TECHNICAL FIELD

The current disclosure relates to the preparation of food products comprising seaweed, in particular meat and meat substitutes comprising seaweed.

BACKGROUND OF THE INVENTION

There is general consensus that the large-scale consumption of meat has several negative impacts on the environment. There is an increasing awareness of the environmental burden of meat consumption, therefore market shares of meat alternatives are growing rapidly.

Seaweeds are an attractive ingredient for hybrid meat products, e.g. products containing both meat and significant amount of plant-based ingredients, and meat substitutes, e.g. food products resembling meat products but that do not contain animal muscle protein. First, the use of seaweed in meat (substitute) products may reduce environmental burden. Seaweed grows in seawater and does not require scarce resources such as farmland and fresh water or pollutants such as fertilizers and pesticides. Moreover, seaweed absorbs CO2 and therefore has a negative carbon footprint. Second, the use of seaweed can lead to improved food products in terms of taste and health. Seaweed comprises functional fibers, therefore requiring less fat for taste experience and mouthfeel. In addition, seaweed is supportive in taste for applications in both sweet and savory food products. Third, seaweed may provide health benefits over the consumption of meat, such as due to the healthier metabolic profile (e.g., lower lipid and cholesterol values).

Despite the aforementioned advantages of seaweed as a food product ingredient, its industrial use has been largely hampered to date. Most seaweed species that are suitable for application as a food ingredient are harvested during spring or early summer. Within hours after harvest the seaweed starts to deteriorate. For quality reasons, seaweed needs to be processed within 48 hours, preferably within 24 hours.

The moment seaweed is harvested it may comprise around 90% water, which requires it to be dried before distribution to the markets or consumers (Djaeni et al. Low Temperature Seaweed Drying Using Dehumidified Air. Procedia Environmental Sciences, Volume 23, 2015, Pages 2-10). Drying of seaweed withing hours after harvest is considered the “gold- standard” preservation method for seaweeds intended for application in meat (substitute) products. Drying is known to slow down physical and biochemical (autolytic) processes in the deterioration of seaweed, and to limit the spoilage by microorganisms. Upon application in food products the dried seaweed is rehydrated before it is applied as an ingredient for meat (substitute) products. Nevertheless, decrease in sensory profile is observed in dried seaweed (Xiren Guli Keyimu. Elimination of seaweed odour and its effect on antioxidant activity. AIP Conference Proceedings 1614, 399, 2014). Use of dried and rehydrated seaweed leads to unsatisfactory changes in the color and taste profile of the food product. These unsatisfactory changes in color and taste profile may increase upon longer storage of the dried seaweed. Color and taste are among the most important factors for the attractiveness and acceptability of meat and meat substitute food products, and an inappropriate and/or inconstant color and/or taste profile is usually perceived as (microbial) food spoilage by the consumer. In order to improve the attractiveness of seaweed-based food products and/or mask the loss in sensory (color and/or taste) profile, only limited amounts can commonly be introduced into the food product.

There is an unmet need in the production of seaweed-based meat (substitute) food products (in particular when comprising relatively large amount of seaweed) that are attractive to the consumer, and which do not show a loss in their sensory profile. For this reason, the amount of seaweed that can be incorporated into a food product, without having negative effects on product quality and sensory profile, is considered to be limited. The current invention provides a solution to one or more of the above-mentioned problems.

SUMMARY OF THE INVENTION

The inventors found that the use of freshly frozen seaweed improves the sensory profile and shelf-life of both refrigerated and freshly frozen meat (substitute) products comprising seaweed. The improvement in sensory profile and shelf-life becomes particularly advantageous when increasingly high amounts of seaweed are used in a meat (substitute) product and/or when the storage or preservation time of the seaweed increases. The improvement in the sensory profile and shelf-life with the use of freshly frozen seaweed is especially evident when compared to the use of dried (and rehydrated) seaweed. The use of dried seaweed did not lead to satisfactory meat (substitutes) and was not compatible with high amounts of seaweed (e.g. 15 wt.% or more, preferably 20 wt.% or more).

The inventors discovered the following features of food products comprising freshly frozen seaweed (different from food products comprising dried seaweed) that are considered to contribute to an improvement in sensory profile and/or shelf-life:

- a prevention of browning in the form of so-called melanosis of the seaweed ingredient

- a reduction of discoloration in the form of “methemoglobin formation” of a meat product - an improvement in shelf life (e.g., due to improved antimicrobial and anti-oxidative activity)

- a reduction in fishy taste and/or fishy odour

- an improvement in cooking weight yield/ lower cooking loss of the food product (e.g., due to reduced water and/or fat loss during cooking)

- an improvement in juiciness and tenderness (e.g., due to reduced water and/or fat loss during cooking, higher water holding capacity).

Resulting from the previous, the inventors discovered that the use of freshly frozen seaweed may render meat (substitute) products with a better health profile, because improvement in sensory profile, shelf life and cooking yield reduces the need of (unhealthy) food additives.

Improved anti-microbial properties of the freshly frozen seaweed as compared with the dried seaweed is an important benefit for a customer. Increased shelf life is thanks to inhibition of growth of food-spoiling and/or pathogenic bacteria and thereby potentially is a cost-safer for retailer and consumer as ‘best-before’ dates on pack can possibly be extended. The antimicrobial effect of freshly frozen seaweed may reduce or eliminate the need of antimicrobial compounds (e.g. E200-E299 and/or E700-E799).

Improved cooking yield is an important benefit for a customer, for example a meat processor or a meat substitute producer, as there is less raw product of the meat (substitute) needed for the same amount of end product, which reduces input of raw materials per unit weight of end product.

In addition, or alternatively, the improved cooking yield, which is related to juiciness and tenderness (e.g., due to reduced water and/or fat loss during cooking, higher water holding capacity), reduces the need to add sodium chloride as a water holding capacity enhancer and/or flavouring agent as most of the taste of the food product is in the water and fat. Currently, (partial) meat substitutes rely on relatively high salt content and additives (for example E-number ingredients like phosphates and methylcellulose, E461) to enhance water and fat holding capacity, and induce and enhance flavor/taste, tenderness, and/or juicy mouthfeel of the finished food product after preparation. However, concerns are rising worldwide about high sodium levels in consumer diets. Also, commonly applied food additives such as phosphates and methylcellulose (E461) are increasingly scrutinised for their potential adverse health effects.

Aforementioned features and advantages were not that clearly observed with the use of dried (and rehydrated) seaweed according to the prior art. Moreover, high amounts of freshly frozen seaweed can be used in meat (substitute) products (e.g. >15 wt.% or more), while maintaining desirable sensory profile (foremost color, taste, smell, juiciness, tenderness) and shelf-life.

In an aspect, the current disclosure relates to the use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, for preparing a food product comprising seaweed in an amount of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%, wherein the food product is a meat product or a meat substitute product.

In an aspect, the current disclosure relates to a use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, for preventing or reducing color change in a food product, wherein the color change is preferably due to metmyoglobin and/or melanosis, wherein the food product is a meat product or a meat substitute product, wherein the food product comprises seaweed in an amount of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%.

In an aspect, the current disclosure relates to a use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, for decreasing weight loss of a food product during cooking, wherein the decrease in weight loss of the food product during cooking is preferably determined by heating 100 g of the food product in an oven providing a temperature of 130 °C and 10% air humidity, and measuring weight loss when the food product has reached a core temperature of 72 °C, wherein the food product is a meat product or a meat substitute product, wherein the food product comprises seaweed in an amount of 1- 50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%.

As used herein, the term “air humidity” means the concentration of water vapour present in the surrounding air. The “air humidity” herein is preferably the relative humidity in the surrounding air, often expressed as a percentage (the ratio of the actual amount of water vapour present in the volume of air at a given temperature to the maximum amount that air would hold at that temperature).

In an aspect, the current disclosure relates to a use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, for increasing juiciness of a food product, wherein the juiciness is defined by a decrease in weight loss during cooking and/or an increase in water holding capacity, wherein the decrease in weight loss of the food product during cooking is preferably determined by heating 100 g of the food product in an oven providing a temperature of 130 °C and 10% air humidity, and measuring weight loss when the food product has reached a core temperature of 72 °C, wherein the food product is a meat product or a meat substitute product, wherein the food product comprises seaweed in an amount of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%.

In an aspect, the current disclosure relates to a use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, for increasing tenderness of a food product, wherein an increase in tenderness is preferably measured by a decrease in peak force (N/mm) as determined for a round core (obtained from the food product) with a diameter of 1.27 cm subjected to a shear test using a Warner Bratzler cutter shearing at a speed of 200 mm/min using a 10-kg load cell, wherein the food product is a meat product or a meat substitute product, wherein the food product comprises seaweed in an amount of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%.

In an aspect, the current disclosure relates to a use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, for increasing shelf-life of a food product (preferably a refrigerated food product) comprising seaweed in an amount of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%, wherein the food product is a meat product or a meat substitute product.

In an aspect, the current disclosure relates to a use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, to prevent or reduce microbial growth in food product comprising seaweed in an amount of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%, wherein the food product is a meat product or a meat substitute product.

In an aspect, the current disclosure relates to a use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, for reducing freezer burn in a food product (preferably a frozen food product) comprising seaweed in an amount of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%, wherein the food product is a meat product or a meat substitute product.

In an aspect, the current disclosure relates to a use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, as (partial) substitute in a food product for one or more selected from the group consisting of an antimicrobial compound, antioxidant regulator, an acidity regulator, an emulsifier, a stabilizer, and a thickening agent, wherein the food product is a meat product or a meat substitute product, wherein the food product comprises seaweed in an amount of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%. In an aspect, the current disclosure relates to a use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, seaweed for reducing fishy taste and/or fishy odour of a food product comprising seaweed in an amount of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%, wherein fishy taste and/or fishy odour is preferably measured by trimethylamine (TMA) content and/or total volatile bases (TVBs) content in the food product, wherein the food product is a meat product or a meat substitute product.

In an aspect, the current disclosure relates to a method of preparing a food product comprising seaweed, preferably Himanthalia elongata seaweed, in an amount of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%, the method comprising: a) providing freshly frozen seaweed, preferably Himanthalia elongata seaweed, and optionally thawing the freshly frozen seaweed; and b) combining the seaweed obtained in step a) with one or more food components to obtain the food product, wherein the food product is a meat product or a meat substitute product.

In an aspect, the current disclosure relates to a food product comprising seaweed, preferably Himanthalia elongata seaweed, in an amount of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%, wherein the food product is a meat product or a meat substitute product, wherein the food product is obtainable by a method comprising: a) providing freshly frozen seaweed, preferably Himanthalia elongata seaweed, and optionally thawing the freshly frozen seaweed; and b) combining the seaweed obtained in step a) with one or more food components to obtain the food product, wherein the food product is a meat product or a meat substitute product.

DETAILED DESCRIPTION OF THE INVENTION

In an aspect, the current disclosure relates to a of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, for preparing a food product comprising seaweed in an amount of preferably 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%, wherein the food product is a meat product or a meat substitute product.

The inventors found that the current invention allows for high amounts of seaweed to be incorporated into meat (substitute) products, while maintaining desirable sensory profile (e.g. color, taste, smell, tenderness, juiciness) and shelf-life. This allows the skilled person to prepare food products with various amounts of seaweed. In an embodiment, the food product herein is brought to the customer and/or the consumer cooled and/or refrigerated through retail, wherein the freshly frozen seaweed may predominantly provide an antimicrobial effect and/or an anti-discoloration effect as disclosed herein in the food product.

In an embodiment, the food product herein is brought to the customer and/or the consumer frozen and/or in a freezer through retail or wholesale, wherein the freshly frozen seaweed may predominantly provide for an anti-freezer burn and/or an anti-discoloration effect as disclosed herein in the food product.

The food product comprising seaweed as disclosed herein may comprise seaweed in an amount of at least 1, or 5, or 10, 11, 12, 13, 14, or 15, 16, 17, 18, 19, or 20, 21, 22, 23, 24, or 25, or 30, or 35, or 40, or 45, or 50, or 55, or 50, or 55, or 60, or 65, or 70, or 75, or 80, or 85, or 90, or 95 wt.%. In addition, or alternatively, the food product comprising seaweed as disclosed herein may comprise seaweed in an amount no more than 95, or 90, or 85, or 80, or 75, or 70, or 65, or 60, or 55, or 50, or 45, or 40, or 35, or 30, or 25, or 20, or 15, or 10, or 5 wt.%.

As used herein, the term “seaweed” may mean any species of seaweed (i.e. macroalgae) or a combination (e.g. blend, mixture) of two or more types of species of seaweed, such as belonging to one or more of the group of three major groups of seaweed: brown seaweed (brown algae, Phaeophyceae), green seaweed (green algae, Chlorophyta), and red seaweed (red algae, Rhodophyta). As used herein, the term “seaweed” encompasses the "kelps”, meaning large brown algae seaweed that make up the order Laminariales. Preferably the “seaweed” as disclosed herein is Himanthalia Elongata or a combination (e.g. blend, mixture) Himanthalia Elongata with one or more other species of seaweed, preferably Undaria pinnatifida, Palmaria Palmata and/or Chondrus Crispus. The seaweed as disclosed herein is preferably a red or brown seaweed (brown algae). Regarding red seaweeds, e.g. one or more of: Chondrus Crispus, Palmaria Palmata. Regarding brown seaweeds: e.g. one or more of: Eisenia bicyclis, Alaria esculenta, Durvillaea antarctica, Ecklonia cava, Saccharina japonica, Laminaria digitata, Postelsia palmaeformis, Nereocystis luetkeana, Saccharina latissima, Undaria pinnatifida, Undaria undarioides, Fucus vesiculosus, Pelvetia canaliculate, Sargassum fusiforme, Sargassum echinocarpum, Sargassum cinetum, Sargassum vulgare, Sargassum swartzii, Sargassum myriocysum, Fucus spiralis, Himanthalia elongata, and Cladosiphon okamuranus. In an embodiment, the food product comprising seaweed as disclosed herein further comprises Himanthalia Elongata in combination with one or more other species of seaweed, preferably Undaria pinnatifida, Palmaria Palmata and/or Chondrus Crispus.

In an embodiment, the food product comprising seaweed as disclosed herein further comprises only Himanthalia Elongata as a species of seaweed.

As used herein “freezing” of seaweed means subjecting the seaweed to a temperature of below 0 °C, preferably of below -2 °C, more preferably of below -15 °C, wherein said freezing is preferably performed within 48 or, more preferably, 24 hours after harvest. In addition, or alternatively, “freezing” as used herein preferably means that the seaweed is subjected to said reduced temperature for at least 24 hours, preferably at least one week, more preferably at least one month, even more preferably at least one year. The term “freshly frozen seaweed” as used herein means that the seaweed is, or has been, subjected to freezing, wherein the freezing preferably has a purpose preservation and/or storage. The term “freshy frozen” preferably encompasses that the seaweed has been subjected to freezing within 72 hours, preferably 48 hours, more preferably within 24 hours, after harvest. In the current disclosure the term “freshly frozen seaweed” may be used interchangeably with the term “frozen’, “freeze-stored” or “freeze-preserved seaweed”. Preferably, the “freshly frozen” seaweed as disclosed herein has been washed prior to freezing. Preferably, “freshly frozen seaweed” herein refers to a seaweed that is other than being washed and or treated to reduce levels of iodine and/or contaminants such as heavy metals and allergens. Preferably, the “freshly frozen” seaweed herein has not been, subjected to another preservation and/or storage method than freezing (e.g. pickling and/or drying). In an embodiment “freshly frozen seaweed” refers to seaweed that has been harvested (i.e. collected from its origin of growing or cultivation) longer than a day ago (i.e. prior to the use according to the invention), preferably longer than a week ago, more preferably longer than a month ago, most preferably longer than a year ago and/or wherein the seaweed has been subjected to freezing for at least 24 hours (in total), preferably at least one week (in total), more preferably at least one month (in total), most preferably at least one year (in total). The “freezing” preferably refers to freezing without interruption (i.e. the seaweed is not subjected to thawing in between two freezing steps). The “freshly frozen seaweed” as disclosed herein encompasses a seaweed has been thawed once or more during freeze-storage. For example, a seaweed that after harvest has been kept in freshly frozen state (e.g. at -20 °C) for one month, subsequently kept in unfreshly frozen state for one month, and subsequently kept in a freshly frozen state for another month, is considered a "freshly frozen seaweed”, more specifically a “freshly frozen seaweed” subjected (in total) to at least two months of freezing. For example, a seaweed that after harvest has been preserved by freezing (and not subjected to drying for a purpose of preservation), and optionally is thawed for processing into a food product, is herein considered a “freshly frozen seaweed”.

In an embodiment, freshly frozen Himanthalia elongata seaweed is defined as Himanthalia elongata seaweed which has been subjected to freezing within 72 hours, preferably within 48 hours, more preferably within 24 hours, after harvest.

In a preferred embodiment, the “freezing” as disclosed herein does not encompass the use of dry ice or another solid form of gas.

In an embodiment, the seaweed in the food product comprising seaweed as disclosed herein has been harvested a least one day before preparing the food product, preferably one week before preparing the food product, more preferably at least one month before preparing the food product, even more preferably at least one year before preparing the food product, most preferably at least two years before preparing the food product.

In an embodiment, the seaweed used in the preparation of the food product comprising seaweed as disclosed herein has been freshly frozen, and preferably never thawed after its harvest before application.

In the current disclosure the term “drying” of seaweed means that the seaweed is subjected to a process involving reducing moisture content in seaweed, preferably a process aimed at removing most (e.g. >90%, >95%, >99%) of the moisture content from the seaweed. The term “drying” as used herein encompasses among others air drying, heat drying, freeze drying, and sun drying. Preferably, the “term” drying refers to a method which the skilled person considers as preservation method (of seaweed), and which involves drying. As used herein “dried seaweed” encompasses seaweed that is dried and thereafter rehydrated, such as before incorporating into a food product. As used herein, “dried seaweed” can be used interchangeably with “dried an rehydrated seaweed”.

As used herein, a “meat product” means a food product that comprises meat (i.e. flesh of an animal, including flesh of a fish), preferably in an amount of at least 1 wt.%, more preferably in an amount of at least 5 wt.%. In addition, or alternatively, a “meat product” as used herein means a food product wherein a part of the protein (e.g. >50%) is derived from animal muscle protein. In a preferred embodiment, a “meat product is a food product comprising 90% or less than 90%, e.g. 70, 60, 50, 40, 30, 20, 10, 5, 1, or 0 wt.% by weight meat, while preferably having a meat protein content of more than 20, 30, 40, 50, 60, 70 wt.%. A “meat product” as used herein encompasses, among others, a meat burger, a meat steak, a meat fillet, a meat sausage, a meat sausage roll, a meat snack, a meatball, a meat nugget, a meat schnitzel, a meat cutlet, and minced meat. As used herein, a “meat substitute” means a food product that does not contain animal muscle protein. In addition, or alternatively, the term “meat substitute” as used herein refers to a food product which is not a meat product, but preferably with at least some similarities in aesthetics, appearance, texture and/or taste as compared with a meat product. In a preferred embodiment, the protein in a meat substitute is completely non-meat derived. The term “meat substitute” as used encompasses (and may be used interchangeably with), among others, one or more selected from the group comprising a (meat) burger substitute, a (meat) steak substitute, a (meat) fillet substitute, a (meat) sausage substitute, a (meat)sausage roll substitute, a (meat) snack substitute, a meatball substitute, a (meat) nugget substitute, a (meat) schnitzel substitute, a (meat) cutlet substitute, and a minced meat substitute.

In an embodiment, the meat substitute as disclosed herein is a vegetarian food product. In an embodiment, the meat substitute as disclosed herein can be a vegan food product.

The inventors found that the freezing of seaweed leads to less discoloration of the seaweed comprising food product, e.g. as compared to drying of seaweed.

In an aspect, the current disclosure relates to a use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, for preventing or reducing color change in a food product, wherein the color change is preferably due to metmyoglobin and/or melanosis, wherein the food product is preferably a meat product or a meat substitute product, wherein the food product comprises seaweed in an amount of preferably 1-50 wt.%, preferably of IQ- 40 wt.%, more preferably of 15-35 wt.%.

As used herein “melanosis” in a food ingredient/product means a coloration and/or pigmentation that occurs in a polyphenol containing food ingredient/product, typically by oxidation. Generally, melanosis occurs when polyphenols are exposed to ambient air (e.g. during storage), which is enhanced by drying of the food ingredient/product containing polyphenols.

As used herein “metmyoglobin” (metMb) means the oxidized form of myoglobin. Myoglobin stores oxygen in muscle cells, and it therefore found in high amounts in meat and animal- protein-based foods. When a food high in myoglobin is exposed to oxygen, the food typically changes towards a tan or brown colour (often together with pigmentation) when the iron in myoglobin oxidizes.

In a preferred embodiment (e.g., a meat product), the color change due to metmyoglobin and/or melanosis is a (gradual) change in one or more of the L*a*b* values as defined by the International Commission on Illumination (CIE), preferably defined by a decrease in L*, an increase in a*, and/or a decrease in b*.

In an embodiment, the freshly frozen seaweed as disclosed herein and/or the freshly frozen seaweed comprised in the food product as disclosed herein has a colour profile defined by the following L*a*b* values:

- L* in the range of 50-80; and/or

- a* in the range of -15 to +15; and/or

- b* in the range of +45 to +75.

The one or more of the L*a*b values as disclosed herein can be measured by any suitable method known to the skilled person. Widely accepted methods are colorimetry or spectrophotometry. In an embodiment, the L*a*b is measured by colorimetry, using a standard white calibration (e.g. L*=97.10, a*=0.19, b*=1.95), CIE standard illuminant D65, and 10° standard observer angle setting.

The water holding capacity of an uncooked product is directly related to the amount of amount of water and/or fat in an uncooked food product and may be positively associated with the juiciness and tenderness of a cooked food product. In an embodiment, an increase in water holding capacity of an uncooked product is measured by a centrifugal method as disclosed herein.

As used herein, the term “water holding capacity” of an uncooked food product means the ability of the uncooked food product to retain liquid water when subjected to external forces such as compression centrifugation. Typically, the water holding capacity inversely correlates to the amount of liquid water released from a food product subjected to centrifugal force (e.g. as disclosed in Jauregui et al. 1981. A simple centrifugal method for measuring expressible moisture, a water-binding property of muscle foods. J. Food Sci. 46: 1271-1271, Zhang et al. LWT - Food Science and Technology Volume 28, Issue 1, 1995, Pages 50-55). For example, 30 g of a food product is weighed and diced into 1 cm ³ cubes. The cubes are placed in a centrifuge tube over paper with known weight. The tube is centrifuged at 860 x g for 7.5 min at 20 °C in a lab benchtop centrifuge. The weight increase of the paper normalized to the initial weight of the food product relates to the liquid water released from the food product. For example, a higher amount of liquid water released from the food product means a lower water holding capacity. An improved water holding capacity of an uncooked food product enables lowering or leaving out of emulsifiers, stabilizers and/or thickening agents.

The water holding capacity of a cooked food product is directly related to the amount of amount of water and/or fat in a cooked food product and may be positively associated with the juiciness and tenderness of a cooked food product. In an embodiment, an increase in juiciness and tenderness is measured by a decrease in weight loss during cooking.

In an aspect, the current disclose relates to the use of freshly frozen seaweed, preferably freshly frozen Himanthalia elongata seaweed, for decreasing weight loss of a food product during cooking, wherein the decrease in weight loss of the food product during cooking is preferably determined by heating 100 g of the food product in an oven providing a temperature of 130 °C and 10% air humidity, and measuring weight loss when the food product has reached a core temperature of 72 °C, wherein the food product is a meat product or a meat substitute product, wherein the food product comprises seaweed in an amount of 1- 50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%.

In an embodiment, the use of freshly frozen seaweed (e.g. Himanthalia elongata) in the current disclosure is for decreasing the weight loss during cooking of the seaweed and/or of a food product comprising the seaweed, wherein the decrease in weight loss is preferably relative to dried and rehydrated seaweed. In an embodiment, the food product as disclosed herein, preferably obtainable by the method as disclosed herein, has a lower weight loss during cooking than a food product comprising dried and rehydrated seaweed.

In a preferred embodiment, the food product has a weight loss during cooking which is lower as compared to a food product wherein the freshly frozen Himanthalia elongata seaweed in step a) of the method as disclosed herein is replaced by dried and rehydrated Himanthalia elongata seaweed, wherein the weight loss of the food product during cooking is preferably determined by heating 100 g of the food product in an oven providing a temperature of 130 °C and 10% air humidity, and measuring weight loss when the food product has reached a core temperature of 72 °C.

As used herein, the “cooking” refers to any method that is suitable to prepare the food product for consumption and that involves heating, e.g. cooking may be one or more of baking, ovenheating, frying, roasting, grilling, steaming, poaching, simmering, broiling, blanching, braising, and stewing. The cooking protocol generally depends on the food product. Preferably, the cooking protocol follows the cooking instructions as recommended to the consumer by the manufacturer and/or supplier, for example as stated on the packaging of the food product. In a preferred embodiment, the “weight loss during cooking” is determined by heating 100 g of a food product (preferably intact, disc shaped and with a thickness of 1 cm) at 130 °C (preferably in an environment with 10% air humidity), until the food product reaches a core temperature of 72 °C.

The term “weight loss during cooking” as used herein may be used interchangeably with the term “cooking loss”. As used herein, the term “weight loss during cooking” preferably means the water and/or fat loss as a result of cooking. The term “weight loss” may herein be used interchangeably with “water and/or fat loss”.

In a preferred embodiment, the weight loss during cooking (%) is determined with the following equation: Weight loss during cooking (%) = (weight before cooking-weight after cooking)/weight before cooking.

In a preferred embodiment, the cooking yield (%) can be calculated with the following equation: Cooking yield (%)= 100% - weight loss during cooking (%).

The food product as disclosed herein (e.g. meat product or meat substitute product) may have a weight loss during cooking of no more than 30%, or 25%, or 20%, or 15%, or 10%, or 5%. In addition or alternatively, the food product as disclosed herein (e.g. meat product or meat substitute product) may have a weight loss during cooking of at least 5%, or 10%, or 15%, or 20%, or 25%, or 30%.

The inventors consider that a lower weight loss during cooking results in a meat (substitute) product made from freshly frozen seaweed leads to an increased juiciness (mouth meat juice release upon biting), a reduced biting effort (i.e. “increased tenderness”) and a more satisfying mouthfeel during consumption. At first glance, tenderness appears to be simply a measure of the biting effort required. An improved water holding capacity of both in uncooked and cooked state enables lowering or leaving out of emulsifiers, stabilizers and/or thickening agents.

In an embodiment, the current disclosure relates to a use of freshly frozen Himanthalia elongata seaweed for increasing juiciness of a food product, wherein the juiciness is preferably defined by a decrease in weight loss during cooking and/or an increase in water holding capacity, wherein the cooking is preferably by heating 100 g of the food product in an oven at 130 °C, providing 10% moisture, until the food product has reached a core temperature of 72 °C, wherein the food product is a meat product or a meat substitute product, wherein the food product comprises seaweed in an amount of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%.

In an embodiment, an increase in juiciness as disclosed herein is defined by a decrease in weight loss during cooking.

In an embodiment, an increase in juiciness as disclosed herein is defined by an increase in water holding capacity, wherein the water holding capacity is preferably determined on a food product subjected to a method of cooking as disclosed herein.

In an aspect, the current disclosure relates to a use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, for increasing tenderness of a food product, wherein the food product comprises seaweed in an amount of preferably 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%. An increase in in tenderness is preferably measured by a decrease in peak shear force (N) when subjecting the food product to shearing by Warner Bratzler shearing device, more preferably wherein said shearing is performed on round cores (1.27 cm in diameter) obtained from the food product at a speed of 200 mm/min using a 10-kg load cell. In an embodiment, the shearing as disclosed herein uses the following protocol: the food product is allowed to equilibrate to room temperature. A round core (1.27 cm in diameter) of food product is sheared on a Warner Bratzler shearing device (United Testing machine, e.g. United SSTM-500) at a cross head speed of 200 mm/min using a 10-kg load cell, and a 1.02 cm thick V-shape blade (V-shape) with a 60°angle and a half-round peak. The peak force (kg) needed to shear each the samples is recorded, converted to Newtons (N), and the mean peak shear force is reported.

The peak force herein is preferably determined for a food product that is cooked and brought back to room temperature (23°C), wherein the cooking may be any appropriate method as disclosed herein, but preferably involves heating of 100 g of the food product in an oven at 130 °C (e.g. 10% air humidity) until the food product has reached a core temperature of 72

In an aspect, the current disclosure relates to a use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, for increasing shelf-life of a freshly frozen (-1°C to -20°C) and/or cooled (below 7°C) food product comprising seaweed in an amount preferably of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%, wherein the food product is a meat product or a meat substitute product.

The inventors found that the current invention allows for the preparation of food products comprising seaweed with extended shelf-life of refrigerated meat (substitute) products. The shelf-life is typically dependent on the type of food product. The food product comprising seaweed as disclosed herein may have a shelf-life of at least 1 day, or 2 days, or 3 days, or 4 days, or 5 days, or 6 days, or 7 days, or 21 days or 28 days. In addition or alternatively, the food product comprising seaweed as disclosed herein seaweed as disclosed herein may have a shelf-life of no more than 28 days, or 21 days, or 1 month, or 7 days, or 6 days, or 5 days, or 4 days, or 3 days, or 2 days, or 1 day.

The inventors found that the current invention allows for the preparation of food products comprising seaweed with extended shelf-life of frozen meat (substitute) products. The shelflife is typically dependent on the type of food product. The food product comprising seaweed as disclosed herein may have a shelf-life of at least 1 day, or 1 month, or 3 months, or 4 months, or 5 months, or 6 months, or 7 months, or 9 days or 12 months. In addition or alternatively, the food product comprising seaweed as disclosed herein seaweed as disclosed herein may have a shelf-life of no more than 36 months, or 21 months, or 12 months, or 7 months, or 6 months, or 5 months, or 4 months, or 3 months, or 2 months, or 1 month.

The term “shelf-life” of a food product as used herein means the length of time for which said food product remains fit for consumption and/or saleable. In addition, or alternatively, the “shelf-life” means the length of time between production of a food product (e.g. meat product or meat substitute product), preferably comprising seaweed, and the expiration (best before) date of the food product (e.g. as disclosed on the packaging of the food product). Herein, the “shelf-life” may refer to the shelf-life of seaweed (e.g. as food component, as semi-finished food product) and/or the shelf life of a food product, preferably a meat (substitute) product comprising seaweed. In an embodiment, the “shelf life” herein is defined as the best before date on the packaging. In an embodiment, the “increasing shelf life” herein means extending the best before date. In an embodiment, the “shelf-life” refers to the shelf-life of a refrigerated and/or frozen food product.

The inventors found that the use of freshly frozen seaweed in meat (substitutes) extends the shelf-life of the meat (substitute) comprising seaweed (e.g. as compared to use of dried and rehydrated seaweed), without negative effects on the sensory profile. The naturally improved shelf-life eliminates or reduces the need of food additives such as antioxidants and (synthetic) antimicrobial compounds.

It is known in the art that increased antimicrobial activity in meat (substitutes) may help retard the growth of microbes, amongst them food spoiling and/or pathogenic micro-organisms (Papuc et al. Comprehensive Reviews in Food Science and Food Safety. November 2017, Volume16, Issue 6, p.1243 - 1268). In accordance, the inventors consider the features of freshly frozen seaweed may help retard the said food spoilage and/or pathogenic microorganisms, thus providing another mechanism by which shelf-life is improved in meat (substitutes) prepared from freshly frozen seaweed. In addition or alternatively, the inhibition or slowing down of growth of pathogens may reduce the risk of disease caused by intake of foodborne bacteria.

As used herein, the term “food spoilage micro-organism” means a micro-organism, preferably a bacterium, that causes spoilage of food, but which does not necessarily cause lifethreatening infections. In the current disclosure, the following pathogens are considered as food spoilage pathogens: Brochothrix thermosphacta, Carnobacterium spp., Lactobacillus spp., Lactococcus spp., Leuconostoc spp., Pediococcus spp., Stretococcus spp., Kurthia zopfii, and Weisella spp.

As used herein, the term “pathogenic micro-organism” means a micro-organism, preferably a bacterium, that typically can cause life-threatening infections, and furthermore may also contribute to food spoilage. In the current disclosure, the following pathogens are considered as foodborne pathogens: Bacillus cereus, Campylobacter jejuni, Clostridium botulinum, Clostridium perfringens, Cronobacter sakazakii, Esherichia coli, Listeria monocytogenes, Salmonella spp., Shigella spp., Staphylococccus aureus, Vibrio spp. and Yersinia enterocolitica.

In an embodiment, the use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, is for reducing growth of food spoiling micro-organisms and/or pathogenic micro-organisms in a food product comprising seaweed disclosed herein.

Preferably the term “reducing growth” of micro-organisms as used herein means a reduction of at least 1 log, more preferably of at least 2 log, more preferably of at least 3 log, even more preferably of at least 4 log in the number of pathogens in a food product, e.g. in comparison to a food product not comprising seaweed and/or in comparison to a food product comprising dried (and rehydrated) seaweed. In an aspect, the current disclosure relates to a use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, to prevent or reduce microbial growth in a food product comprising seaweed in an amount of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%, wherein the food product is a meat product or a meat substitute product.

In an aspect, the current disclosure relates to a use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, for reducing freezer burn in a food product (preferably a frozen food product) comprising seaweed in an amount of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%, wherein the food product is a meat product or a meat substitute product.

As used herein, the term “freezer burn” means a condition that occurs when water sublimation and water crystal formation increases the oxidation of fatty acids of food products, amongst others due to air reaching deeper into the food product. Freezer burn may appear as an off-taste of the food product, which can be the result from lipid oxidation. Typically, reducing or preventing the freezer burn will improve the flavor and/or color of a frozen food product. In an embodiment, a reduction in freezer burn is defined by a reduction in the level of lipid peroxidation marker, preferably malondialdehyde. The level of the lipid peroxidation marker herein, such as malondialdehyde, can be determined by high-performance liquid chromatography.

In an aspect, the current disclosure relates to a use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, in a food product as substitute or partial substitute for one or more selected from the group comprising an antimicrobial compound, an antioxidant regulator, an acidity regulator, an emulsifier, a stabilizer, and a thickening agent, wherein the food product is a meat product or a meat substitute product, wherein the food product comprises seaweed in an amount of preferably 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%.

In a preferred embodiment, the antimicrobial compound is an (synthetic) antimicrobial compound, more preferably identified by an E (Europe) number according to the International Numbering System for Food Additives, such as one or more of E200-E299 and/or E700- E799.

As used herein, the term “antimicrobial compound” means a compound that reduces growth and/or kills microbes (e.g. bacteria, fungi, virus, preferably bacteria), preferably of one or more food spoilage micro-organisms and/or pathogenic micro-organisms as disclosed herein, wherein the compound is preferably a synthetic compound. The “antimicrobial compound” herein is preferably one or more of an (synthetic) antibiotic compound, an (synthetic) antiviral compound, an (synthetic) antifungal compound and an (synthetic) antiparasitic compound. Preferably, the antimicrobial compounds herein is an antibacterial compound. Natural (food) products, such as seaweed, are herein not considered as antimicrobial compounds. In an embodiment, the antimicrobial compound is a synthetic antimicrobial compound.

In an embodiment, the current disclosure relates to a use of freshly frozen seaweed, preferably freshly frozen Himanthalia elongata seaweed, for decreasing bacterial number in a food product comprising seaweed in an amount of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%, wherein the food product is a meat product or a meat substitute product, wherein the bacterial number is defined in terms of the colony forming units per gram of sample (CFU/g). The skilled person is aware of the methods to determine the CFU.

In a preferred embodiment, the antioxidant regulator and/or the acidity regulator is identified by an E (Europe) number according to the International Numbering System for Food Additives, preferably by one or more of E300-E399, more preferably by E301 (sodium ascorbate) and/or by E330 (citric acid).

In a preferred embodiment, the emulsifier, stabilizer, and/or thickening agent is identified by an E number according to the International Numbering System for Food additives, preferably by one or more of E400-E499, preferably by E400 (alginate) and/or by E461 (methylcellulose).

In an aspect, the current disclosure relates to a use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, as substitute or partial substitute in a food product of one or more of a food colour additive (e.g. one or more of E100- E199), preservative (e.g. E200-E299), and flavour enhancer (e.g. E600-E699), wherein the food product is a meat product or a meat substitute product, wherein the food product comprises seaweed in an amount of preferably 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%.

As used herein, the term “antioxidant regulator” means an ingredient, that is (primarily) used in a food product to reduce oxidation in a food product. Reducing oxidation typically may provide improved taste, colour, and/or shelf-life (over a longer period).

The skilled person is well-aware of ingredients that may serve as antioxidants in food products, for example one or more selected from the group comprising tocopherols, ascorbic acid (e.g. L-ascorbate, sodium L-ascorbate, calcium L-ascorbate, potassium L-ascorbate), rosemary extract, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), tertiary butylhydroquinone (TBHQ), propyl galate, vitamin C, vitamin E, a carotenoid (e.g. beta-carotene), selenium, manganese, glutathione, coenzyme Q10, lipoic acid, a flavonoid, phenol, a polyphenol, lecithin, L-tartaric acid, citric acid, erythorbic acid. The antioxidant regulator as taught herein may or may not additionally serve as a thickening agent, and/or an emulsifier in a food product. The antioxidant as disclosed herein is preferably one or more ingredients as identified by an E number according to the International Numbering System for Food additives, preferably by one or more of E300-E399, preferably by one or more of E300 (ascorbic acid), E301 (sodium ascorbate), E306 (tocopherols), E310 (prolyl glate), E319 (TBHQ), E320 (BHA), E321 (BHT), E392 (rosemary extract).

As used herein, the term “acidity regulator” means an ingredient that is (primarily) used in a food product to change the acidity, preferably increase the acidity, and/or to improve the organoleptic quality by imparting the food product with a different, preferably an increased, acid flavour. The skilled person is well-aware of ingredients that may serve as acidity regulator in food products, for example one or more selected from the group comprising citric acid (E330), acetic acid (E260), lactic acid (E270), fumaric acid (E297), calcium acetate (E263), and malic acid (E296), and combinations thereof. The acidity regulator as disclosed herein may or may not additionally serve as an antioxidant regulator. The acidity regulator as disclosed herein is preferably one or more ingredients as identified by an E number according to the International Numbering System for Food additives, preferably by one or more of E300- E399.

As used herein, the term “stabilizer” means an ingredient, that is (primarily) used in a food product to enhance stability, shelf-life, ease of processing, gel formation, and/or stiffness. The skilled person is well-aware of ingredients that may serve as stabilizer in food products, for example one or more selected from the group comprising starches such as arrowroot, cornstarch, potato starch, sago, tapioca, vegetable gums such as guar gum, xanthan gum, locust bean gum, pectin such as pectin from apples or pectin from citrus fruits, proteins such as collagen, egg whites, gelatin, whey, other such as agar, carrageenan, sodium pyrophosphate, soy lecithin, egg lecithin, monoglycerides and/or diglycerides, and combinations thereof. The stabilizer as taught herein may or may not additionally serve as a thickening agent, and/or an emulsifier in a food product. The stabilizer as disclosed herein is preferably one or more ingredients as identified by an E number according to the International Numbering System for Food additives, preferably by one or more of E400-E499. As used herein, the term “emulsifier” means an ingredient that is (primary) used in a food product as border between two immiscible liquids. The skilled person is well-aware of ingredients that may serve as emulsifier in food products, e.g. one or more selected form the group comprising mustard, soy lecithin, egg lecithin, monoglycerides, diglycerides, polysorbates, carrageenan, guar gum and/or canola oil, and combinations thereof. In the current disclosure, the emulsifier may also serve as a thickening agent. The emulsifier as disclosed herein is preferably one or more ingredients as identified by an E number according to the International Numbering System for Food additives, preferably by one or more of E400- E499.

As used herein, the term “thickening agent” means an ingredient that is (primarily) used in a food product to increase the viscosity. The skilled person is well-aware of ingredients that may serve as thickening agent in food products, e.g one or more selected from the group comprising locust bean gum, carrageenan, seaweed extract, guar gum, xanthan gum, extracellular polysaccharide gum, scleroglucan, agar, pectin, alginate, cellulose derivatives (e.g. methylcellulose), gum tragacanth and/or gum arabic, and combinations thereof. In the current disclosure, the thickening agent may also serve as an emulsifier. The thickening agent as disclosed herein is preferably one or more ingredients as identified by an E number according to the International Numbering System for Food additives, preferably by one or more of E400-E499, preferably by E400 (alginate) and/or by E461 (methylcellulose).

As part of the invention, reduced levels of sodium may be used in the food product comprising seaweed. For example, the meat (substitute) comprising seaweed as disclosed herein may have a sodium content (per kg of food product) of less than 20 g (e.g. 10-20 g), preferably less than 10 g (e.g. 1-10 g), more preferably less than 5 g (e.g. 1 - 5 g), more preferably less than 1 g (e.g. 0.1 - 0.1 g), wherein the meat (substitute) comprising seaweed preferably has a juiciness as defined by a weight loss during cooking as disclosed herein.

As part of the invention, reduced levels of phosphate may be used in the food product comprising seaweed. For example, the meat (substitute) comprising seaweed as disclosed herein may have a phosphate content (per kg of food product) of less than 2 g (e.g. 1-2 g), preferably less than 0.2 g (e.g. 0.1-0.2 g), more preferably less than 0.02 g (e.g. 0.01 - 0.02 g), more preferably less than 0.002 g (e.g. 0.001 - 0.002 g), wherein the meat (substitute) comprising seaweed preferably has a juiciness as defined by a weight loss during cooking as disclosed herein. The phosphate herein is preferably identified by one or more of the group consisting of E338 (Phosphoric acid), E339 (Sodium phosphates), E340 (Potassium phosphates), E341 (Calcium phosphates), E343 (Magnesium phosphates).

The inventors found that the use of a freshly frozen seaweed to prepare a meat(substitute) with seaweed reduces the taste and odour associated with food (fish) spoilage. Herein the “fishy taste” and/or “fishy odour” means a taste or smell, respectively, in a food product that is associated with food (fish) spoilage. In addition, or alternatively, the “fishy taste” and/or “fishy odour” herein may mean a taste or smell, respectively, due to a compound associated with food (fish) spoilage, e.g. one or one or more of ammonia, trimethylamine (TMA, e.g. expressed as trimethylamine nitrogen, TMA-N), dimethylamine (DMA, e.g. expressed as dimethylamine nitrogen, DMA-N), and monomethylamine, known as total volatile basic nitrogen (TVB-N). The term “fishy taste” may herein be used interchangeably with the term “rotten taste”). The term “fishy odour” may herein be used interchangeably with “rotten odour”. In a preferred embodiment, a reduction in fishy taste and/or fishy odour is defined by a reduction in one or more of ammonia, TMA, TMA-N, DMA, DMA-N, monomethylamine, and TVB-N.

In an aspect, the current disclosure relates to a use of freshly frozen seaweed, preferably of freshly frozen Himanthalia elongata seaweed, for reducing or preventing fishy taste and/or fishy odour of a food product comprising seaweed in an amount of preferably1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%, wherein fishy taste and/or fishy odour is preferably measured by trimethylamine (TMA) content and/or total volatile bases (TVBs) content in the food product, wherein the food product is a meat product or a meat substitute product.

In a preferred embodiment, the use of froze seaweed, preferably of freshly frozen Himanthalia elongata seaweed, as disclosed herein is for increasing the antioxidant activity of the food product comprising seaweed.

In a preferred embodiment, the food product comprising seaweed as disclosed herein has an antioxidant activity of 50-90%, preferably 60-90%, expressed as % singlet oxygen radical scavenging capacity.

In the present disclosure, the “antioxidant activity” is preferably defined by the % singlet oxygen radical scavenging capacity, more preferably measured by the 2,2-diphenyl-1- picrylhydrazyl (DPPH) assay (Rajauria et al. Journal of Food Biochemistry 37(3) 2012). For example, the protocol may be (similar to) the following: The assay is performed in a 96-well round-bottomed microtiter plate with 1 :1 (v/v) ratio of 100 mL of DPPH radical solution (165 mM) and 100 mL of sample. Different concentrations are tested for each sample in order to get the % of singlet oxygen radical scavenging capacity. The DPPH solution is freshly prepared for each experiment in methanol. The reaction mixtures are incubated for 30 min at 25 °C in dark conditions, and absorbance is measured at 517 nm in a microtiter plate reader. The ability to scavenge the DPPH radical is calculated using the equation: scavenging capacity (%) = 1- [(A _sam pie - A _sam pie blank)/(A _con troi)], where Acontroi is the absorbance of the control (DPPH solution without sample), A _sam pie is the absorbance of the test sample (DPPH solution plus test sample) and A _sam pie blank is the absorbance of the sample only (sample without any DPPH solution). Calculated values indicate the concentration of sample required to scavenge DPPH radicals (expressed as %). The higher % value of the sample, the higher antioxidant capacity.

The antioxidant activity of the food product comprising seaweed as disclosed herein may be defined by a singlet oxygen radical scavenging capacity (%) of at least 10%, or 20%, or 30%, or 40%, or 50%, or 60%, or 70%, or 80%, or 90%. In addition or alternatively, the antioxidant activity of the food product comprising seaweed as disclosed herein may be defined by a % singlet oxygen radical scavenging capacity (%) of no more than 90%, or 80%, or 70%, or 50%, or 40%, or 30%, or 20%, or 10%. The antioxidant activity of the food product comprising seaweed as disclosed herein is preferably defined by a singlet oxygen radical scavenging capacity (%) of 40-90%, preferably, 50-90%, more preferably 60-90%.

In a preferred embodiment, the freshly frozen seaweed, preferably the freshly frozen Himanthalia elongata seaweed, as disclosed herein is in a composition further comprising one or more ingredients selected from the group consisting of a preservative, a seasoning, a flavor compound, a spice, a herb, a sugar, a pepper, a dextrose, a phosphate, an ascorbic acid, a protein, and a salt.

In an embodiment, the one or more ingredients are mixed with the seaweed when the seaweed is in a freshly frozen state (e.g. at a temperature of -5 °C or less, preferably of -10 °C or less), as to form the composition comprising seaweed and one or more ingredients as disclosed herein.

In a preferred embodiment, the freshly frozen seaweed, preferably the freshly frozen Himanthalia elongata seaweed, as disclosed herein is in a composition further comprising one or more other species seaweed other than Himanthalia elongata, preferably Undaria pinnatifida, Chondrus Crispus, Palmaria Palmata, Saccharina Latissima and/or Alaria Esculenta.

In a preferred embodiment, the Himanthalia elongata seaweed and the one or more species of seaweed other than Himanthalia elongata are provided in the composition as disclosed herein in a weight ratio of 20:1 - 1 :1 , such as 10:1 - 1 :1 or 5:1 - 2:1.

In an aspect, the current disclosure relates to a method of preparing a food product comprising seaweed, preferably Himanthalia elongata, e.g. in an amount of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%, the method comprising: a) providing freshly frozen seaweed, preferably freshly frozen Himanthalia elongata seaweed, and optionally thawing the freshly frozen seaweed; and b) combining the seaweed obtained in step a) with one or more food components to obtain the food product, wherein the food product is a meat product or a meat substitute product.

The term “food component” as used herein encompasses one or more selected from the group comprising meat, fish, a vegetable, a plant, an (animal) fat, an oil, a plant protein, a nutritional compound, a preservative, a seasoning, a flavor compound, a spice, a herb, a sugar, a pepper, a dextrose, a phosphate, an ascorbic acid, and a salt.

The plant protein as disclosed herein may be derived from a pulse, an oil seed, tuber, grain, or is pea protein, soy protein, rapeseed protein, potato protein, bean protein, lupine protein.

The nutritional compound’ as disclosed herein may be any ingredient that may act to enhance the nutritional value of a food product, to restore any nutrients lost or degraded during production of a food product, to enrich a food product in order to correct dietary deficiencies, and/or to add nutrients to food substitutes. The skilled person is well-aware of ingredients that may serve as nutritional compound in a food product, such as a vitamin (e.g. vitamin A, B, C, and/or D), an essential fatty acid (e.g. linoleic acid, cholesterol, omega-3 fatty acid, oleic acid, omega-6 fatty acid, docosahexaenoic acid, polyunsaturated fatty acid, a mineral (e.g. calcium, iron), and/or dietary fibre.

In an embodiment, the combining in step b) of the method as disclosed herein is by one or more methods selected of the group comprising mixing, homogenizing, blending, cutting, folding, kneading, sifting, stirring, beating, and creaming. In an embodiment, the seaweed is at a temperature of between -250 °C and 5 °C, preferably of between -10 °C to 0 °C when combining with the one or more other food components in step b) of the method as disclosed herein.

In a preferred embodiment, the method of preparing a food product comprising seaweed as disclosed herein comprises: a) providing freshly frozen Himanthalia elongata seaweed and optionally thawing the freshly frozen Himanthalia elongata seaweed; and b) combining the Himanthalia elongata seaweed obtained in step a) with one or more food components to obtain the food product.

In a preferred embodiment, the seaweed in step a) and/or the seaweed in step b) has a moisture content of 25-99%, preferably 50-99%, more preferably 75-99%. In an embodiment, the moisture content of the seaweed in step a) and/or the seaweed in step b) is at least 50%, or 60%, or 70%, or 80%, or 90%.

In an aspect, the current disclosure relates to a food product comprising seaweed, preferably Himanthalia elongata seaweed, e.g. in an amount of 1-50 wt.%, preferably of 10-40 wt.%, more preferably of 15-35 wt.%, wherein the food product is a meat product or a meat substitute product, wherein the food product is obtainable by a method comprising: a) providing freshly frozen seaweed, preferably Himanthalia elongata seaweed, and optionally thawing the freshly frozen seaweed; and b) combining the seaweed obtained in step a) with one or more food components to obtain the food product, wherein the food product is a meat product or a meat substitute product.

In a preferred embodiment, the food product comprising seaweed as disclosed is obtainable by a method comprising: a) providing freshly frozen Himanthalia elongata seaweed and optionally thawing the freshly frozen Himanthalia elongata seaweed; and b) combining the Himanthalia elongata seaweed obtained in step a) with one or more food components to obtain the food product.

In a preferred embodiment, the food product comprising seaweed as disclosed herein has a juiciness which is higher compared as compared to a food product wherein the freshly frozen Himanthalia elongata seaweed in step a) is replaced by dried, and optionally rehydrated, Himanthalia elongata seaweed, wherein the juiciness is defined by a decrease in weight loss during cooking and/or an increase in water holding capacity, wherein the cooking is preferably by heating 100 g of the food product in an oven at 130 °C, providing 10% moisture, until the food product has reached a core temperature of 72 °C.

In a preferred embodiment, the food product comprising seaweed as disclosed herein has a bacterial number which is lower as compared as compared to a food product wherein the freshly frozen Himanthalia elongata seaweed in step a) is replaced by dried, and optionally rehydrated, Himanthalia elongata seaweed, wherein the bacterial number is preferably defined by the colony forming units per gram of food product (CFU/g).

In a preferred embodiment, the food product comprising seaweed as disclosed herein has a bacterial number that is at least 6 log, 5 log, 4 log, 3 log, 2 log, or 1 log lower as compared as compared to a food product wherein the freshly frozen Himanthalia elongata seaweed in step a) is replaced by dried, and optionally rehydrated, Himanthalia elongata seaweed, wherein the bacterial number is preferably by the colony forming units per gram of food product (CFU/g).

The food product comprising seaweed as disclosed herein may have a bacterial number of no more than 10 log CFU/g food product, such as more than 9, 8, 7, 6, 5, 4, 3, 2, 1 log CFU/g food product.

In a preferred embodiment, the food product comprising seaweed as disclosed herein has a color as defined by:

- a higher L* as compared to a food product wherein the freshly frozen seaweed (e.g. Himanthalia elongata in step a) is replaced by dried, and optionally rehydrated seaweed (e.g. Himanthalia elongata), and/or

- a higher a* as compared to a food product wherein the freshly frozen seaweed (e.g. Himanthalia elongata in step a) is replaced by dried, and optionally rehydrated seaweed (e.g. Himanthalia elongata)’, and/or

- a lower b* as compared to a food product wherein the freshly frozen seaweed (e.g. Himanthalia elongata in step a) is replaced by dried, and optionally rehydrated seaweed (e.g. Himanthalia elongata), wherein the L*a*b* values are as defined by the CIE.

In a preferred embodiment, the food product comprising seaweed as disclosed herein does not comprise an antimicrobial compound and/or comprises an antimicrobial compound in an amount of no more than 1 mg/kg food product, preferably no more than 0.1 mg/kg, more preferably no more than 0.01 mg/kg, even more preferably no more than 0.001 mg/kg. In a preferred embodiment, the food product comprising seaweed as disclosed herein does not comprise an antioxidant regulator and/or comprises an antioxidant regulator in an amount of no more than 1 mg/kg food product, preferably no more than 0.1 mg/kg, more preferably no more than 0.01 mg/kg, even more preferably no more than 0.001 mg/kg.

In a preferred embodiment, the food comprising seaweed as disclosed herein does not comprise an acidity regulator and/or comprises an acidity regulator in an amount of no more than 1 mg/kg food product, preferably no more than 0.1 mg/kg, more preferably no more than 0.01 mg/kg, even more preferably no more than 0.001 mg/kg.

In a preferred embodiment, the food comprising seaweed as disclosed herein does not comprise an emulsifier and/or comprises an emulsifier in an amount of no more than more than 50 g per kg food product, preferably no more than more than 10 g per kg food product, more preferably no more than 1 g per kg food product, even more preferably no more than 0.1 g per kg food product, most preferably no more than 0.1 g per kg food product.

In a preferred embodiment, the food comprising seaweed as disclosed herein does not comprise a stabilizer and/or comprises a stabilizer in an amount of no more than more than 50 g per kg food product, preferably no more than more than 10 g per kg food product, more preferably no more than 1 g per kg food product, even more preferably no more than 0.1 g per kg food product, most preferably no more than 0.1 g per kg food product.

In a preferred embodiment, the food comprising seaweed as disclosed herein does not comprise a thickening agent and/or comprises a thickening agent in an amount of no more than more than 50 g per kg food product, preferably no more than more than 10 g per kg food product, more preferably no more than 1 g per kg food product, even more preferably no more than 0.1 g per kg food product, most preferably no more than 0.1 g per kg food product.

In a preferred embodiment, the antioxidant regulator and/or the acidity regulator as disclosed herein is identified by an E number according to the International Numbering System, preferably one or more of E300-E399, more preferably by E301 (sodium ascorbate) and/or by E330 (citric acid).

In a preferred embodiment, the stabilizer and/or thickening agent as disclosed herein is identified by an E number according to the International Numbering System, preferably by one or more of E400-E499, preferably by E400 (alginate) and/or by E461 (methylcellulose). The inventors found that compounds related to (causing) fishy taste and/or fishy odour are reduced in meat (substitute) product prepared with freshly frozen seaweed. Ammonia, TMA, TMA-N, DMA, DMA-N, monomethylamine, and TVB-N are herein considered as “compounds related to (causing) fishy taste and/or fishy odour”.

The food product comprising seaweed as disclosed herein may comprise one or more compounds relating to fishy taste and/or fishy odour in an amount of no more than 100 mg, or 10 mg, or 8 mg, or 7 mg, or 6 mg, or 5 mg, or 4 mg, or 3 mg, or 2 mg, or 1 mg, or 0.5 mg, or 0.1 mg, or 0.05 mg or 0.01 mg, or 0.001 mg, wherein the amount is the amount per kg of food product or the amount per kg seaweed comprised in the food product. In addition or alternatively, the food product comprising seaweed as disclosed herein may comprise one or more compounds relating to fishy taste and/or fishy odour in an amount of at least 0.001 mg, or 0.01 mg, or 0.05 mg, or 0.1 mg, or 0.5 mg, or 1 mg, or 2 mg, or 3 mg, or 4 mg, or 5 mg, or 6 mg, or 7 mg, or 8 mg, or 9 mg or 10 mg, or 100 mg, wherein the amount is the amount per kg of food product or the amount per kg seaweed comprised in the food product.

In a preferred embodiment, the food product as disclosed herein comprises

- TMA and/or TVBs in an amount of no more than 50 mg/kg, preferably no more than 5 mg/kg, more preferably more than 0.5 mg/kg, wherein the amount is the amount in the food product; and/or

- TMA and/or TVBs in an amount of no more than 200 mg, preferably of no more than 20 mg, more preferably of no more than 2 mg, wherein the amount is the amount per kg seaweed in the food product.

For quantitative measurement of TMA and TVB-N, the Conway dish (Microdiffusion dish) method is preferably used. In this method, TMA and TVB are determined as their nitrogen (TMA-N and TVB-N respectively). The optimal method may be dependent on the type of food product, but the preferred protocol is as follows (based on Ng, C. S. (1987). In H. Hasegawa (Ed.), Laboratory Manual on Analytical Methods and Procedures for Fish and Fish Products, pp. B-3.1-B-3.8):

Reagents a) Inner ring solution — 1% boric acid solution containing indicator:

Take 10 g of boric acid in 1 liter flask, add 200 ml of ethanol. After dissolving boric acid, add 10 ml of mixed indicator solution, then make up to 1 liter with distilled water. b) Mixed indicator solution: Dissolve bromocresol green (BCG) 0.01 g and methyl red (MR) 0.02 g in 10 ml of ethanol. c) 0.02N HCI:

Dilute 20 ml of 1N HCI standard solution with distilled water and make up to 1000 ml. d) Saturated K2CO3 solution:

Take 60 g of potassium carbonate (K2CO3), and add 50 ml of distilled water. Boil gently for

10 min. After cooling down, obtain filtrate through filter paper. e) 50% K2CO3 solution:

Dilute saturated K2CO3 solution twice with distilled water. f) 4% trichloroacetic acid (CCI3COOH) (TCA) solution:

Dissolve 40 g of TCA in 960 ml of distilled water. g) Sealing agent:

Take 3 g of Tragacanth gum, add 30 ml of distilled water, 15 ml of glycerine and 15 ml of 50% saturated K2CO3 solution and mix well. h) Neutralized 10% formaldehyde solution:

Add 10 g of MgCCh to 100 ml of formalin (35% formaldehyde solution) and shake in order to neutralize the acidity of formalin. Filter and dilute filtrate 3 times with distilled water. i) 1% TiCh aqueous solution:

Take 6.7 ml of 15% TiCh solution into 100 ml volumetric flask and make up to 100 ml with distilled water. j) Saturated KNO3 aqueous solution:

Dissolve about 55 g of KNO3 in 50 ml of distilled water.

Apparatus

Conway’s unit:

Wash with detergent (use neutral detergent if available), then rinse with running water and leave until dry. Do not wipe with cloth.

Micro- burette

Procedure

A. Sample extraction

1. Take 2 g of minced sample.

2. Add 8 ml of 4% TCA solution and grind well.

3. Stand for 30 min at ambient temp, with occasional grinding. 4. Filter through filter paper (Whatman No. 41). (or Centrifuge at 3000 rpm, for 10 min.)

5. Keep the filtrate in -20°C freezer if necessary.

B. Determination of TVB-N

1. Apply sealing agent to Conway’s unit.

2. Pipette 1 ml of inner ring solution into inner ring.

3. Pipette 1 ml of sample extract into outer ring.

4. Slant the Conway’s unit with cover.

5. Pipette 1 ml of saturated K2CO3 solution into outer ring.

6. Close the unit.

7. Mix gently.

8. Stand for 60 min at 37°C in incubator.

9. Titrate inner ring solution with 0.02 N HCI using a micro-burette until green colour turns to pink.

10. Do blank test using 1 ml of 4% TCA instead of sample extract.

C. Determination of TMA-N

Principle of TMA-N determination is similar to TVB-N determination except addition of formaldehyde to the sample solution. Formaldehyde is added in order to fix any ammonia present in the sample.

1. Apply sealing agent to Conway’s unit.

2. Pipette 1 ml of inner ring solution into inner ring.

3. Pipette 1 ml of sample extract into outer ring.

4. Pipette 1 ml of neutralized 10% formaldehyde into outer ring.

5. Slant the Conway’s unit with cover.

6. Pipette 1 ml of saturated K2CO3 solution into outer ring.

7. Close the unit.

8. Mix gently.

9. Stand for 60 min. at 37°C in incubator.

10. Titrate inner ring solution with 0.02N HCI using a micro-burette until green colour turns to pink.

11. Do blank test using 1 ml of 4% TCA instead of sample extract.

The terms ‘comprising’ or ‘to comprise’ and their conjugations, as used herein, refer to a situation wherein said terms are used in their non-limiting sense to mean that items following the word are included, but items not specifically mentioned are not excluded. It also encompasses the more limiting verb ‘to consist essentially of’ and ‘to consist of’. Reference to an element by the indefinite article ’a’ or ‘an’ does not exclude the possibility that more than one of the elements is present, unless the context clearly requires that there be one and only one of the elements. The indefinite article ‘a’ or ‘an’ thus usually means ‘at least one’.

The terms ‘to increase’ and ‘increased level’ and the terms ‘to decrease’ and ‘decreased level’ refer to the ability to significantly increase or significantly decrease or to a significantly increased level or significantly decreased level. Generally, a level is increased or decreased when it is at least 5%, such as 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% higher or lower, respectively, than the corresponding level in a control or reference. Alternatively, a level in a sample may be increased or decreased when it is statistically significantly increased or decreased compared to a level in a control or reference.

FIGURE LEGENDS

Figure 1. Images of burgers prepared with 15 wt.% Himanthalia elongata seaweed. Burgers prepared with freshly frozen seaweed (left) retain a lighter and natural red colour and are more generally more appealing. Burgers prepared with dried and rehydrated seaweed (right) are unnaturally darker in colour with brownish colour tone. In addition, burgers prepared with dried and rehydrated seaweed show signs of melanosis, as indicated by the arrows.

Figure 2. Images of meatballs prepared with 15 wt.% Himanthalia elongata seaweed and cooked by baking. Cooked meatballs prepared with freshly frozen seaweed (left) have a reduced cooking loss as compared to cooked meatballs prepared with dried and rehydrated seaweed (right), which contributes to their high juiciness and/or tenderness. Cooked meatballs prepared with dried and rehydrated seaweed (right) are darker in colour and show more signs of oxidation (melanosis).

Figure 3. Images of burgers prepared with 15 wt.% Himanthalia elongata seaweed and cooled at 4 °C for 7 days. Burgers prepared with freshly frozen seaweed (left) retain a lighter and natural red colour and are more generally more appealing. Burgers prepared with dried and rehydrated seaweed (right) show reduction of microbial growth and related spoilage. Burgers prepared with dried and rehydrated seaweed are also unnaturally darker in colour with brownish colour tone. In addition, burgers prepared with dried and rehydrated seaweed show signs of melanosis, as indicated by the arrows. This indicates that freshly frozen Himanthalia elongata seaweed retards microbial growth. EXAMPLES

The following Examples illustrate the different embodiments of the invention.

Example 1

Example 1 shows the taste and sensory profiles in burgers comprising seaweed subjected to different preservation methods.

Freshly harvested Himanthalia elongata seaweed is washed and subjected to one of the two preservation methods for 1 month or 6 months:

1. Drying and rehydration*

2. Freezing at -20 °C

* In the “drying and rehydration group”, the seaweed is air dried in a single layer at 45 °C for 8 hours to remove moisture. The dried seaweed is stored in closed packaging in the dark at room temperature and rehydrated before use.

Burgers are prepared with the following composition:

- 7.5-35% seaweed

- 54-81.5% meat;

- 1.5% starch;

- 1.0% herbs;

- 0.6% NaCI;

- 7.9% water.

The burgers are packed and stored at -20 °C for 1 month. The burgers are cooked in an oven (130 °C, 10-20% air humidity) until a core temperature of 72 °C is reached.

The cooked burgers are presented to panelists. The panelists are asked to score for taste, juiciness, tenderness, and appearance, as being either “very bad”, “average”, “good”, “very good”.

Panelists are also asked to score several undesirable sensory traits typically associated with seaweed-based foods. The panelists use a 10-point scoring system for “fishy taste”, “rotten odor”, and “discoloration” (high color intensity, inhomogeneous color profile). In the 10-point scoring system, 10 means a ‘very unpleasant’ sensory experience and 0 means a ‘very pleasant’ sensory experience.

Table 1 shows the overall taste and sensory profile of burgers comprising 15 wt.% Himanthalia elongata. Table 1 shows that using freshly frozen seaweed which is stored at -20 °C provides a good overall taste and sensory profile of a food product. Even when the seaweed is preserved for 1 month, there is a surprisingly large improvement in the sensory profile for freshly frozen seaweed as compared with drying and rehydration of seaweed. Freezing seaweed at -20 °C for 6 months yields burgers that retain their excellent sensory profile, i.e. being scored as “very good”.

Table 2 shows the presence of undesirable sensory traits typically associated with seaweed, for burgers comprising 15 wt.% Himanthalia elongata. As can be seen, drying can be considered an inappropriate method for amounts of 15% dried seaweed and more. These methods lead to strong fishy taste, rotten odor and unwanted discoloration, especially when the preservation time is prolonged. Especially for drying, the burgers show an overall darker color and inhomogeneous color appearance (denoted as “spotty” appearance” and “brown spots”). There is an absence in fishy taste, rotten odor and discoloration (“spotty” appearance and “brown spots”) when seaweed is freshly frozen as compared to the other preservation methods. When freezing at -20 °C, the preservation time can be increased to at least 12 months without causing undesirable sensory traits.

Table 1. Overall taste and sensory profile of burgers comprising 15 wt.% Himanthalia elongata as scored by panellists (scoring as “very bad”, “bad”, “average”, “good”, “very good”)

Table 2. Scoring of fishy taste, rotten odour, and unpleasant in burgers comprising 15 wt.% Himanthalia elongata. Scoring is done by panellists using a 10-point scoring system (10 means a ‘very unpleasant’ sensory experience; 0 means a ‘very pleasant’ sensory experience).

Example 2

Example 2 shows the effect of the preservation method of seaweed on the presence of undesirable sensory traits in food products, when the amount of seaweed is steadily increased.

Freshly harvested Himanthalia elongata seaweed is washed and subjected to drying/rehydration or freezing at -20 °C for 6 months as described in Example 1. Burgers are prepared the same way as described Example 1 , wherein the wt.% of seaweed is varied at 7.5, 15, and 35 wt.%. The burgers are packed and stored at -20 °C for 1 month. Cooked burgers are scored by panelists for undesirable sensory traits typically using the 10-point scoring system as described in Example 1.

Table 3 shows the presence of undesirable sensory traits typically associated with seaweed, for burgers comprising 7.5, 15, or 35 wt.% Himanthalia elongata. As can be seen, preservation of seaweed at -20 °C allows a higher amount of seaweed to be incorporated in a food product (i.e. at least 35 wt.%), without leading to undesirable sensory traits typically associated with sea-based foods. At 35 wt.%, the burgers comprising freshly frozen seaweed have little “fishy taste”, “rotten odour” and no unwanted discoloration (“spotty” appearance and “grey spots”). This leads to food products that are perceived as having overall very good sensory profile. Overall, drying limits the amount of seaweed that can be incorporated in a food product. At a weight% of 15% and 35%, drying results into a strong fishy taste and rotten odor of the food product.

Table 3. Scoring of fishy taste, rotten odour, and unpleasant in burgers comprising 15 wt.% Himanthalia elongata. The Himanthalia elongata is preserved for 6 months by drying or freezing before use in the burgers. Scoring is done by panellists using a 10-point scoring system (10 means a ‘very unpleasant’ sensory experience; 0 means a ‘very pleasant’ sensory experience).

The inventors consider that appropriate freezing of different species of seaweed leads to improved food products (as shown in Table 1-3), however Himanthalia elongata are thought to provide optimal results. Himanthalia elongata is therefore the preferred seaweed.

Example 3

The present inventors consider that the minimal discoloration in food products comprising freeze-stored seaweed (illustrated in Table 2 and Table 3) is the result of a reduced oxidation in the food product in comparison to the other preservation methods. The reduced oxidation is expected to be the result of an improved antioxidant activity in food products comprising freeze-stored seaweed.

Here, it is illustrated how different preservation methods of seaweed differently affect the formation of metmyoglobin following incorporation into food products.

Figure 1 shows images of burgers prepared with 15 wt.% Himanthalia elongata seaweed (composition and preparation according to Example 1). Burgers prepared with freshly frozen seaweed (left) retain a lighter and natural red colour and are more generally more appealing. Burgers prepared with dried and rehydrated seaweed (right) are losing redness with changes in colour towards a brownish colour tone. The effects become increasingly apparent with increasing the wt.% of seaweed in the burger. Burgers prepared using freshly frozen seaweed are appealing even when up to 50 wt.% seaweed is used.

The Cl ELAB (CIE L* a* b*) color space diagram represents quantitative relationship of colors on three axes:

- L* is represented on a vertical axis with values from 0 (black) to 100 (white). - a* value indicates red-green component of a color, where a* (positive) and a* (negative) indicate red and green values, respectively.

- b* value indicates yellow-blue component of a color, where b* (positive) and b* (negative) indicate yellow and blue values, respectively.

It is determined that the freshly frozen Himanthalia elongata has the following L*a*b* values:

- L* in the range of 50-80 (lower in dried and rehydrated Himanthalia elongata)

- a* in the range of -15 to +15 (higher in dried and rehydrated Himanthalia elongata)

- b* in the range of +45 to +75 (lower in dried and rehydrated Himanthalia elongata)

For the prepared burgers, the actual (absolute) L*a*b are dependent on further ingredients and the wt.% seaweed used.

For the prepared burgers, the L* is typically higher for burgers prepared with freshly frozen seaweed, compared to burgers prepared with dried and rehydrated seaweed;

For the prepared burgers, the a* is typically lower for burgers prepared with freshly frozen seaweed, compared to burgers prepared with dried and rehydrated seaweed;

For the prepared burgers, the b* is typically higher for burgers prepared with freshly frozen seaweed, compared to burgers prepared with dried and rehydrated seaweed.

We herein use a method based on a protocol suggested by the American Meat Science association (page 40, Meat Color Measurement Guidelines, originally established at the 44 ^th Proceedings of the Reciprocal Meat Conference, Revised December 2012) to provide the % Metmyoglobin Formation in the food products. The correlation between the discoloration score (presented in Table 2 and Table 3), the % Metmyoglobin Formation, and the antioxidant activity follows from Table 4.

Table 4. Correlation between the discoloration score, the % Metmyoglobin Formation, and the antioxidant activity.

The putative differences in antioxidant activity can be determined by quantitative methods. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay (Rajauria et al. Journal of Food Biochemistry 37(3) 2012) is a preferred method for this purpose, with the following protocol:

The assay is performed in a 96-well round-bottomed microtiter plate with 1 :1 (v/v) ratio of 100 mL of DPPH radical solution (165 mM) and 100 mL of sample. Different concentrations are tested for each sample in order to get the % of singlet oxygen radical scavenging capacity. The DPPH solution is freshly prepared for each experiment in methanol. The reaction mixtures are incubated for 30 min at 25 °C in dark conditions, and absorbance is measured at 517 nm in a microtiter plate reader. The ability to scavenge the DPPH radical is calculated using the equation: scavenging capacity (%) = 1- [(A _sa mpie - A _S ampie blank)/(A _C ontroi)], where Acontroi is the absorbance of the control (DPPH solution without sample), A _sa mpie is the absorbance of the test sample (DPPH solution plus test sample) and A _sam pie blank is the absorbance of the sample only (sample without any DPPH solution). Calculated values indicate the concentration of sample required to scavenge DPPH radicals (expressed as %). The higher % value of the sample, the higher antioxidant capacity.

Table 5 shows the percentage % metMb formation and antioxidant activity in burgers comprising 7.5, 15, or 35 wt.% Himanthalia elongata (composition and preparation according to Example 1). As can be seen, drying leads to a percentage metMb of 40% or higher, associated with only average antioxidant activity. The percentage metMb is even further increases when the preservation time is prolonged (low antioxidant activity). In comparison, the amount of metMb is 1-20% after both 1 and 6 months of storage, and is associated with a very high antioxidant activity.

Table 5. Percentage metmyoglobin (% metMb) and antioxidant activity in burgers comprising 15 wt.% Himanthalia elongata. The % metMb is based on the discoloration as established by panellists.

Table 6 shows the % metMb formation and antioxidant activity in burgers comprising 7.5, 15, or 35 wt.% Himanthalia elongata (composition and preparation according to Example 1). As can be seen, preservation of seaweed at -20 °C allows a higher amount of seaweed to be incorporated in a food product (i.e. at least 35 wt.%), without leading to high a high amount of metmyoglobin after prolonged preservation. Even at 35 wt.%, the burgers comprising freshly frozen seaweed have low metmyoglobin content and very high antioxidant activity.

Table 6. Percentage metmyoglobin (% metMb) and antioxidant activity in burgers comprising 15 wt.% Himanthalia elongata. The Himanthalia elongata is preserved for 6 months by drying or freezing before use in the burgers. The % metMb is based on the discoloration as established by panellists.

It is expected that similar effects as presented in Table 5 and Table 6 are established by quantitative determination of metMb formation. Suitable methods include colorimetry to measure tristimulus values (CIE L*a*b*), or spectrophotometers that supply spectral analysis for the calculation of tristimulus values.

As shown in Figures 4-6 collectively, the inventors found that melanosis is severe when seaweed is dried. The inventors consider that the melanosis in dried seaweed is due to the accumulation of melanin. In contrast, the inventors found that a discoloration due to melanosis is low or not observed at all when freshly frozen seaweed is applied to a meat product. Also, the typical browning over time of a refrigerated food product due to metmyoglobin formation (i.e. oxidized myoglobin) is reduced when compared to when dried and rehydrated seaweed is applied.

The present inventors consider that the increased antioxidant activity and/or decreased oxidation in freshly frozen seaweed-based meat (substitutes) may offer additional advantages:

- Although metmyoglobin formation and melanosis is not necessarily linked to food spoilage, the effects on the appearance of a food product may impact the consumer decision to purchase and consume the product. The formation of metMb and melanosis may be prevented in freshly frozen seaweed-based meat (substitutes).

- The shelf-life of a food product is dependent, at least in part, on the extent of lipid oxidation (Ruben Dominiguez et al. Antioxidants Basel. 2019 Oct; 8(10): 429). Freshly frozen seaweedbased meat (substitutes) may have an extended shelf-life.

- The high antioxidant activity in freshly frozen seaweed-based meat (substitutes) may reduce or eliminate the need for (synthetic) food additives that are otherwise required to protect the food from spoilage. For instance, antioxidants are often added to inactivate free radicals or metal. Acidity regulators help establish antioxidant activity, but also reduce microbial growth and contribute to food taste/color through pH control. Preservatives may also help in inhibition of microbial growth. The high antioxidant activity of food products based on freeze-stored seaweed may eliminate food additives such as those identified by an E number according to the International Numbering System (e.g. E300-E399- “Antioxidants and acidity regulators”). Food products comprising substances identified by E-numbers are typically considered by consumers as unnatural, artificial and unhealthy.

- An increased antimicrobial activity of freshly frozen seaweed may help retard the growth of pathogens, particularly food spoilage bacteria and/or pathogenic bacteria. Thus, high antimicrobial activity of freshly frozen seaweed may limit bacterial growth and thereby improve shelf-life and safety of the food product. The inventors consider that appropriate freezing of different species of seaweed leads to improved food products (as shown in Table 4-6), however Himanthalia elongata are thought to provide optimal results. Himanthalia elongata is therefore the preferred seaweed.

Example 4

The panellists overall scored the products prepared with freshly frozen seaweed (at -20 °C) as being the juiciest and most tender. The inventors confirmed that foods prepared with freshly frozen seaweed have a higher water and/or fat holding capacity, for example as demonstrated by the reduced loss cooking loss (i.e. loss of water and/or fat during cooking).

The inventors tested the cooking loss that occurs in meatballs prepared according to Example 1 (100 g total weight of meatballs, 15 wt.% seaweed, freezing at -20 °C). In addition, the inventors tested the cooking loss for raw meatballs, and for meatballs pre-cooked burgers (in an oven unit, as could be sold commercially). The pre-cooked meatballs were packed and stored in the fridge prior to use for the cooking loss measurements.

The weight loss during cooking (i.e. cooking loss) (%) was determined with the following equation:

Weight loss during cooking (%) = (weight before cooking-weight after cooking)/weight before cooking.

In the current Example, oven heating was performed in an oven set at 130 °C (10-20% air humidity), until the burgers reached a core temperature of approximately 72 °C.

In the current Example, baking was performed using a defined protocol until the burgers reached a core temperature of approximately 72 °C.

It was found that the meatballs prepared with freshly frozen seaweed have a cooking loss of 23% and 24% after oven-heating an baking, respectively. It was found that the meatballs prepared with dried and rehydrated seaweed have a cooking loss of 28% and 30% after oven-heating an baking, respectively. Thus, in the current Examples, the meatballs prepared with freshly frozen seaweed have approximately 5-6% less cooking loss as compared to meatballs prepared with dried and rehydrated seaweed.

Figure 2 shows representative images of meatballs prepared with 15 wt.% Himanthalia elongata seaweed and cooked by baking. Cooked meatballs prepared with freshly frozen seaweed (left) have a reduced cooking loss as compared to cooked meatballs prepared with dried and rehydrated seaweed (right), which contributes to their high juiciness and/or tenderness. Cooked meatballs prepared with dried and rehydrated seaweed (right) are unnaturally darker in colour and show more signs of oxidation.

The inventors demonstrated the reduced cooking loss for meatballs prepared with 15 wt.% freshly frozen seaweed. Even higher reduction in cooking loss is overall seen when increasing the seaweed content (wt.%).

The inventors consider that appropriate freezing of different species of seaweed improves cooking (water and/or fat) yield, however Himanthalia elongata are thought to provide optimal results. Himanthalia elongata is therefore the preferred seaweed.

Example 5

Autolytic and/or enzymatic activity are/is known to increase the levels of several basic nitrogenous compounds, such as trimethylamine (TMA), and dimethylamine (DMA), together known as total volatile basic nitrogen (TVB-N) in seaweed and seafood. These compounds are most responsible for the unpleasant "fishy tase” and “rotten odor” of several marine food products including seaweed.

Considering the improved sensory profile of food products made from freshly frozen seaweed, the inventors hypothesized that the freezing of seaweed strongly inhibits or delays the formation of TMA and/or TVB-N in food products comprising the seaweed. In agreement, the inventors established that use of freshly frozen seaweed reduces TMA and TVB-N in burgers by approximately three-fold as compared to the use of dried and rehydrated seaweed (Figure 7). The burgers were prepared with 15 wt.% Himanthalia elongata as described in Example 1.

Table 7 illustrates the quantitative TMA and TVB-N levels for the comparison between burgers comprising dried-rehydrated or freshly frozen seaweed. Seaweed-based burgers are prepared as described in Example 1 with 15 wt.% Himanthalia elongata and subsequently kept at 4°C for 7 days. For quantitative measurement of TMA and TVB-N, the Conway dish (Microdiffusion dish) method is used. In this method, TMA and TVB are determined as their nitrogen (TMA-N and TVB-N respectively). The optimal method may be dependent on the type of food product, but the preferred protocol is as disclosed herein (based on Ng, C. S. (1987). In H. Hasegawa (Ed.), Laboratory Manual on Analytical Methods and Procedures for Fish and Fish Products, pp. B-3.1-B-3.8).

Example 6

Acidity regulators are commonly used in meat products, hybrid meat products and meat substitutes in order to lower the pH of the food product. Acidity regulators primarily inhibit bacterial growth, but appropriate pH level is also needed for proper binding functionality of meat proteins. The current inventors found that the use of freshly frozen seaweed reduces or eliminates the need of acidity regulators. As proof-of-concept, the inventors compared the pH of burgers (as prepared in Example 1) either comprising or not comprising the acidity regulator E330 (citric acid).

Table 8 provides the pH of burgers comprising dried-rehydrated or freshly frozen Himanthalia elongata, either or not comprising the acidity regulator E330 (citric acid). When E330 is used, burgers prepared with dried and rehydrated seaweed on average have a higher pH than burgers prepared with freshly frozen seaweed. In absence of E330, burgers prepared with dried and rehydrated seaweed show pH levels unfit for meat (substitute) products, whereas this is not seen for burgers prepared with from freshly frozen seaweed.

Table 8 shows the pH of burgers comprising dried-rehydrated or freshly frozen Himanthalia elongata, and either or not comprising acidity regulator E330 (citric acid). Seaweed-based burgers are prepared as described in Example 1 with 30 wt.% Himanthalia elongata,

The use of E300 (vitamin C) led to similar results as shown in Table 8.

Example 7 The inventors found that a use of frozen Himanthalia elongata seaweed retards microbial growth, whereas this is not seen with a use of dried and rehydrated Himanthalia elongata seaweed.

Figure 3 shows images of burgers prepared with 15 wt.% Himanthalia elongata seaweed as described in Example 1 and cooled at 4 °C for 7 days. Burgers prepared with freshly frozen seaweed (left) retain a lighter and natural red colour and are more generally more appealing. Burgers prepared with dried and rehydrated seaweed (right) show microbial growth and related spoilage. Burgers prepared with dried and rehydrated seaweed are also unnaturally darker in colour with brownish colour tone. In addition, burgers prepared with dried and rehydrated seaweed show signs of melanosis, as indicated by the arrows. This indicates that there is freshly frozen Himanthalia elongata seaweed retards microbial growth.

Thus, it was found that the use of freshly frozen Himanthalia elongata seaweed reduces the bacterial growth (e.g. defined by the colony-forming-units) in the food products. As a result, the inventors consider that the antimicrobial effect of freshly frozen seaweed may reduce or eliminate the need of antimicrobial compounds that are otherwise required to protect the food from spoilage. These include antimicrobial compounds identified by an E number according to the International Numbering System, such as one or more E200-E299 and/or of E700-E799.