CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application is a continuation-in-part of U.S. Patent Application No. 16/010,306, filed June 15, 2018, which claims the benefit of U.S. Provisional Patent Application No. 62/668,656, filed May 8, 2018. This application also claims the benefit of U.S. Provisional Patent Application No. 62/781 ,519, filed December 18, 2018. All of these applications are incorporated herein by reference in their entireties.

TECHNICAL FIELD

[0002] This disclosure relates to seaweed meal and a method of making the same. The seaweed meal disclosed herein can replace the use of agar. The method and the seaweed meal production can be used in the field of food industry.

BACKGROUND

[0003] Agar, also called Kanten in Japan. It is a polysaccharide extracted from red algae such as Gelidium and Gracilaria, and is one of the most widely used algae colloids. Agar has been used as food for several hundred years in China and Japan. It first appeared in the form of jelly-like block, which form is still available nowadays. After World War II Western countries began to industrialize the production of agar powder. Agar powder was initially used for bacteria culturing, later extended to food, cosmetics and pharmaceutical industries. Agar has unique gel performance and gel stability, and therefore, has a wide range of applications in food industry, as well as in pharmaceutical, cosmetics and biological engineering industries.

[0004] Clean Label and disruptive Green have become more and more popular in food industrials, and have been listed in the“Top 10 Innova’s 2017 Trends,” resulting in strong interests in naturalness and clean label. This trend has been growing globally for the past couple of years and is the new standard in the industry. It provides vast market space development potential for seaweed meal to replace agar in food application.

[0005] US Patent Application Publication No. 2015/0164125 describes a method for making seaweed meal, including the steps of harvesting the seaweed from a sea floor; chopping the seaweed; cleaning the seaweed; desiccating the seaweed; grinding the seaweed to a meal; drying the seaweed between the cleaning step and the desiccating step, the drying step is accomplished by a seaweed agitator. US Patent No. 4,125,392 describes a seaweed filter cake product produced by conditioning raw seaweed by mixing it with water and other substances, and then digesting the conditioned seaweed by cooking it with steam to form a steam-digested mixture. The filter cake is skimmed off the top of the resulting steam-digested mixture and used by itself, or as a starting product for producing fertilizers or plant growth stimulators in liquid or pelletized form.

[0006] The prior-art references merely produced a simple seaweed meal or cake products, with poor gelling capability, brown or green color, and strong fishery flavor. These prior-art seaweed products are not suitable for use as gelling agent or stabilizer in food applications to replace agar extracted from red seaweed. Thus, there is a need in the field to produce better quality seaweed meal suitable for food applications.

SUMMARY

[0007] In one aspect, provided herein is a method of making a seaweed meal to replace agar in the food industry. The method comprises the steps of cleaning the fresh or dried red seaweed, pretreating the red seaweed with an alkali, pre-treating the red seaweed with an acid, treating the red seaweed with an enzyme such as a cellulase, and drying and grinding the treated red seaweed to obtain the seaweed meal. Optionally, the method comprises a step of bleaching the red seaweed with a bleaching agent before the drying and grinding step. In some embodiments, the bleaching step is carried out before the enzyme treatment step. In some embodiments, the bleaching step is carried out after the enzyme treatment step. The disclosure provides a simple process to produce a seaweed meal with light color, good flavor and taste, good mouthfeel, high gelling capability, great stability, and ideal smoothness and slipperiness suitable for“clean label” food applications.

[0008] In a related aspect, provided herein is a seaweed meal produced by the method described above. The seaweed meal produced by the disclosed method can be used in food industry to replace agar as gelling ingredient in, for example, dairy, jelly, pudding applications.

BRIEF DESCRIPTION OF THE DRAWINGS

[0009] This application contains at least one drawing executed in color. Copies of this application with color drawing(s) will be provided by the Office upon request and payment of the necessary fees.

[0010] Figures 1A-1 C shows the FTIR results of seaweed meal Sample 2 (top curve in each panel) in comparison to commercially available seaweed powder (Sample 10) (middle curve in each panel), and agar (Sample 11 ) (bottom curve in each panel). Figure 1A shows the entire range while Figures 1 B and 1 C show the selective ranges (enlarged). The spectral feature at approximately 890 cm ¹ is mainly associated with the C-H bending at the anomeric carbon in b-galactose residues (Figure 1 C). The presence of a strong band in the FTIR spectra at 930 cm ^-1 is indicative of the occurrence of 3,6- anhydro-D-galactose (Figure 1 C). And 1550-1510 cm ^-1 is associated with protein N-H bending (amide bands II) (Figure 1 B). The seaweed meal obtained by the process disclosed herein (Sample 2) contains lower protein (Figure 1 B) and higher 3,6-anhydro- D-galactose (Figure 1 C) than the commercially available seaweed powder (Sample 10). In comparison to the commercially available agar (Sample 11 ), Sample 2 contains slightly higher protein (Figure 1 B) but slightly lower b-galactose and 3,6-anhydro-D- galactose (Figure 1 C).

DETAILED DESCRIPTION

[0011] Methods for making a high quality seaweed meal suitable for food applications are provided herein. The method is a simple process to produce a seaweed meal with desired features such as light color, good flavor and taste, good mouthfeel, high gelling capability, great stability, and ideal smoothness and slipperiness suitable for food applications. The seaweed meal produced by the disclosed method can be used in food industry to replace agar as a gelling agent in, for example, producing dairy, jelly, pudding products. [0012] In some embodiments, the method disclosed herein comprises the steps of cleaning seaweed material with water, pre-treating the clean seaweed material with an alkali, pre-treating the alkali-treated seaweed material with an acid, bleaching the pre-treated seaweed material with a bleaching agent, and treating the bleached seaweed material with an enzyme to obtain a seaweed meal suitable for food applications. The bleaching step is optional and can be included if a light color seaweed meal is desired. In some embodiments, the method further comprises the step of processing the enzyme-treated seaweed by twin Screw Extruded process, and then crushing and drying the extruded seaweed.

[0013] Seaweed is also called macroalgae, marine algae, or seagrass. The seaweed material used for the disclosed method includes red algae, for example, Gracilaria, Gelidium, Porphyra, and Ahnfeltia. In some embodiments, a combination of at least two, at least three, or at least four of the disclosed red algae can be used.

[0014] The seaweed material is cleaned by water prior to any treatment. Seaweed materials in any form such as dry seaweed, fresh seaweed, or rehydrated seaweed, can be used. It is known in the art that fresh seaweed can be dehydrated to obtain dry seaweed. The dry seaweed can be rehydrated by soaking the dry seaweed in water. In some embodiments, the dry seaweed is rehydrated during the cleaning step. When dry seaweed is used, the dry seaweed:water ratio in the cleaning step can be between 1 :1 and 1 :40, between 1 :3 and 1 :30, between 1 :5 and 1 :25, between 1 :5 and 1 :20, between 1 :10 and 1 :20, or between 1 :5 and 1 :10; when fresh seaweed or rehydrated seaweed is used, the fresh seaweed:water ratio in the cleaning step can be between 1 : 1 and 1 :3, between 1 : 1 and 1 :5, between 1 :2 and 1 :5, between 1 :1 and 1 :10, or between 1 : 1 and 1 :15.

[0015] The alkali that can be used for the disclosed method includes sodium hydroxide, potassium hydroxide, calcium hydroxide, or a combination of these alkalis. The alkalis in one or several forms (e.g. various hydrates) can be used in this disclosure. In the pre-treatment step, the seaweed:alkali ratio can be between 1 :1 and 1 :3, between 1 :1 and 1 :5, between 1 :2 and 1 :5, or between 1 :1 and 1 :10. The alkali solution can be at a concentration of between 1 % and 10%, between 3% and 15%, between 5% and 15%, between 5% and 10%, between 10% and 20%, between 3% and 20%, or between 10% and 30% (wt%). In some embodiments, the alkali pre-treatment step is performed at a temperature between 40°C and 80°C, between 65°C and 95°C, between 50°C and 100°C, or between 70°C and 120°C. In some embodiments, the alkali pre-treatment can be performed for a period of between 0.5 hour and 24 hours, between 0.5 hour and 20 hours, between 0.5 hour and 15 hours, between 0.5 hour and 12 hours, between 0.5 hour and 8 hours, between 0.5 hour and 6 hours, between 1 hour and 4 hours, between 1.5 hours and 3 hours, or between 0.5 hour and 4 hours.

[0016] The acid that can be used for the disclosed method includes hydrochloric acid, nitric acid, phosphate acid, and oxalic acid. Alternatively, a combination of the disclosed acids, such as a combination of at least two acids, at least three acids, or at least four acids, can be used. In the pre-treatment step, the seaweed:acid ratio can be between 1 :1 and 1 :3, between 1 :1 and 1 :5, between 1 :2 and 1 :5, or between 1 :1 and 1 :10. The acid solution can be at a concentration of between 0.05% and 5%, between 0.1 % and 3%, between 0.1 % and 2%, between 0.1 % and 1 %, or between 0.1 % and 0.5% (wt%). In some embodiments, the acid pre-treatment step can be performed for a period of between 5 minutes and 240 minutes, between 10 minutes and 180 minutes, between 10 minutes and 120 minutes, between 10 minutes and 60 minutes, or between 20 minutes and 60 minutes.

[0017] Optionally, the seaweed is treated with a bleaching agent to obtain a light color seaweed meal. The bleaching agent that can be used for the disclosed method includes hypochlorous acid, sodium hypochlorous, and chlorine dioxide. Alternatively, a combination of the disclosed bleaching agents, such as a combination of at least two bleaching agents, or at least three bleaching agents, can be used. In the bleaching step, the seaweed: bleaching agent ratio can be between 1 :1 and 1 :3, between 1 :1 and 1 :5, between 1 :2 and 1 :5, or between 1 :1 and 1 :10. The bleaching agent can be at a concentration of available chlorine content of between 0.01 % and 5%, between 0.02% and 3%, between 0.05% and 2%, between 0.05% and 1 %, or between 0.05% and 0.5% (wt%). In some embodiments, the bleaching step can be performed for a period of between 5 minutes and 240 minutes, between 10 minutes and 180 minutes, between 10 minutes and 120 minutes, between 10 minutes and 60 minutes, or between 20 minutes and 60 minutes.

[0018] In some embodiments, the pre-treated or bleached seaweed has an impurity of the red seaweed of less than 5%, less than 4%, less than 3%, less than 2%, or less than 1 %, a sulfate content of less than 3%, less than 2%, less than 1 %, or less than 0.5%, and a color of light brown to light yellow.

[0019] The enzyme treatment catalyzes the hydrolysis of the cellulose and/or related polysaccharides or the like in the pre-treated and/or bleached seaweed such that the obtained seaweed meal has an improved mouthfeel. Various enzymes may be used to achieve this goal, for example, cellulase, hemicellulase, pectinase, amylase, xylanase, pentosanase, glucanase, esterase. In some embodiments, one or more enzymes can be used to treat the seaweed. An example of the suitable enzyme is cellulase, including acid cellulase, neutral cellulase, and alkali cellulase, including but not limited to 1 ,4-p-D-glucan glucanohydrolase, endo-1 ,4-p-D-glucanase, 1 ,4-p-D- glucan cellobilhydrolase, exo-1 ,4-p-D-glucannase, b-1 ,4-glucosidase, from Trichoderma, Aspergillus or Penicillium. In some embodiments, a combination of at least two, at least three, or more of these cellulases can be used. In the enzyme treatment step, the seaweed:enzyme ratio can be between 1 :1 and 1 :3, between 1 :1 and 1 :5, between 1 :2 and 1 :5, or between 1 :1 and 1 :10. The enzyme can be at a concentration of between 10 U/g and 500 U/g, between 20 U/g and 400 U/g, between 30 U/g and 300 U/g, between 50 U/g and 200 U/g, or between 75 U/g and 150 U/g. In some embodiments, the enzyme treatment step can be performed for a period of between 0.5 hour and 12 hours, between 0.5 hour and 8 hours, between 0.5 hour and 6 hours, between 1 hour and 3 hours, or between 1 hour and 4 hours, at a pH range of between 2.0 and 10.0, between 3.0 and 9.0, between 3.0 and 8.0, between 4.0 and 7.0, or between 5.0 and 6.0, or at a temperature between 15°C and 100°C, between 20°C and 90°C, between 30°C and 80°C, between 20°C and 70°C, between 30°C and 60°C, or between 40°C and 50°C. [0020] A variety of acids or alkalis can be used to adjust pH. For example, the alkali for pH adjustment is selected from the group consisting of sodium hydroxide, potassium hydroxide, and a combination thereof. The acid for pH adjustment is selected from the group consisting of hydrochloric acid, nitric acid, phosphate acid, and a combination of at least two, or at least three of these acids.

[0021] Optionally, the method disclosed herein further includes processing the enzyme treated seaweed with a twin Screw Extruded process, which can be performed at a temperature between 70°C and 200°C, between 80°C and 150°C, between 100°C and 150°C, between 100°C and 140°C, between 120°C and 140°C, or between 1 10°C and 130°C. The twin Screw Extruded process can be performed in the absence or in the presence of water. For example, the seaweed:water ratio can be between 1 :0 and 1 :5, between 1 : 1 and 1 :3, or between 1 :0 to 1 :2.

[0022] In some embodiments, the clean seaweed is pretreated with 5%-15% (wt%) of an alkali solution at a ratio of between 1 :2 and 1 :5 and at a temperature of between 65°C and 95°C, pretreated with 0.1 %-0.5% (wt%) of an acid solution at a ratio of between 1 :2 and 1 :5, treated with a cellulase having a concentration of between 75 U/g and 250 U/g at a ratio of between 1 :2 and 1 :5 and at a temperature of between 40°C and 60°C, and then treated with a twin Screw Extruded process at a temperature between 1 10°C and 140°C.

[0023] The enzyme treated seaweed or the twin Screw Extruded processed seaweed can be crushed and dried to obtain the seaweed meal. In some embodiments, the dried seaweed meal has a water gel strength of at least 50 g/cm ² , at least 75 g/cm ² , at least 100 g/cm ² , at least 120 g/cm ² , or at least 150 g/cm ² . In some embodiments, the dried seaweed meal has a color of light brown to light yellow. In some embodiments, the dried seaweed meal has a melting point of less than 100°C, less than 90°C, less than 85°C, less than 75°C, or less than 60°C.

[0024] In some embodiments, the seaweed meal obtained by the disclosed technology contains the following ingredients: > 40% soluble dietary fiber, about 20%- 35% insoluble dietary fiber, > 75% total dietary fiber, < 2% sulfate content, and < 5% protein. In some embodiments, the seaweed meal obtained by the disclosed technology has the following properties: gel strength at 1.5% >100 g/cm ² , and a melting point at 1.0% 60-95 °C.

[0025] The following examples are intended to illustrate various embodiments of the invention. As such, the specific embodiments discussed are not to be construed as limitations on the scope of the invention. It will be apparent to one skilled in the art that various equivalents, changes, and modifications may be made without departing from the scope of invention, and it is understood that such equivalent embodiments are to be included herein. Further, all references cited in the disclosure are hereby incorporated by reference in their entirety, as if fully set forth herein.

EXAMPLES

Example 1 : Materials and Methods

[0026] The algae materials used in the experiments are as follows:

[0027] Other reagents include: hypochlorous acid, sodium hypochlorous, available chlorine contents 10%, food grade; chlorine dioxide, available chlorine contents 50%, food grade; sodium hydroxide, potassium hydroxide, calcium hydroxide, 99% food grade; hydrochloric acid, 36%, food grade; and sulfuric acid, nitric acid, oxalic acid, phosphate acid, 99%, food grade; acid cellulase, neutral cellulose, alkaline cellulose, 50,000 U/g, food grade. HAAKE Process 11 Parallel Twin Screw Extruder was used in certain processes.

Example 2: Preparation of Seaweed Meal from Dried Gracilaria

[0028] 10 kg dried Gracilaria was mixed with 50 kg water in the tank, stirred at 60

RPM for 30 minutes to clean the seaweed, and then the water was removed to obtain 50 kg clean seaweed. The clean seaweed was mixed with 50 kg 3% sodium hydroxide solution in the reaction tank, the mixture was heated and the temperature was kept at 50°C, with stirring at 60 RPM for 30 minutes, followed by washing the seaweed with 500 kg water. Once the water was removed, 48 kg of alkali pre-treated seaweed was obtained. The alkali pre-treated seaweed was mixed with 50 kg of 0.1 % hydrochloric acid solution in the tank, stirred for 10 minutes, washed by 100 kg water, and then the water was removed to obtain 46 kg acid pre-treated seaweed. The pre-treated seaweed was mixed with 50 kg 0.05% bleaching agent, hypochlorous acid (HCIO) solution, stirred at 60 RPM for 10 minutes, then the hypochlorous acid solution was removed, and the seaweed was washed with 100 kg water. Once the water was removed 46 kg pre-treated seaweed was obtained. The specification of the pre-treated seaweed was: impurity of the red seaweed < 0.5%, the sulfate content of the seaweed < 0.3%, and the color was yellow.

[0029] Subsequently, the pre-treated seaweed was mixed with 46 kg of 30 U/g acid cellulase solution, pH was adjusted to 3.0 using hydrochloric acid, and the temperature was kept at 20°C, with stirring at 60 RPM for 30 minutes. Then the cellulase solution was removed and the seaweed was washed with 100 kg water. 40 kg cellulase treated seaweed was obtained.

[0030] The seaweed was processed by HAAKE Process 11 Parallel Twin Screw Extruder with a heating temperature set at 100°C. And then the extruded seaweed was crushed and dried to obtain 3.5 kg seaweed meal. Example 3: Preparation of Seaweed Meal from Fresh Gracilaria

[0031] 50 kg fresh Gracilaria was mixed with 150 kg water in the tank, and stirred at 60 RPM for 30 minutes to clean the seaweed. The water was removed to obtain 49 kg clean seaweed. The clean seaweed was mixed with 120 kg of 6% sodium hydroxide solution in the reaction tank. The mixture was heated and the temperature was kept at 85°C, with stirring at 60 RPM for 120 minutes. The seaweed was washed with 600 kg water, and the water was removed to obtain 48 kg alkali pre-treated seaweed. The alkali pre-treated seaweed was mixed with 120 kg of 0.38% phosphate acid solution in the tank and stirred for 30 minutes, and then washed with 100 kg water. The water was removed to obtain 48 kg acid pre-treated seaweed. The acid pre-treated seaweed was mixed with 120 kg 0.1 % bleaching agent, sodium hypochlorous acid solution, stirred at 60 RPM for 30 minutes, the sodium hypochlorous acid solution was removed, and followed by washing with 100 kg water. The water was removed to obtain 45 kg pre- treated seaweed. The specification of the pre-treated seaweed was: impurity of the red seaweed < 0.8%, the sulfate content of the seaweed < 0.2%, and the color was light yellow.

[0032] The pre-treated seaweed was mixed with 120 kg of 100 U/g cellulase solution, pH was adjusted to 5.0 with hydrochloric acid, heated and the temperature was kept at 50°C, with stirring at 60 RPM for 120 minutes. The cellulase solution was removed and the seaweed was washed with 100 kg water. 41 kg of cellulase treated seaweed was obtained.

[0033] The treated seaweed was processed by HAAKE Process 11 Parallel Twin Screw Extruder with heating temperature set at 128° C. And then the extruded seaweed was crushed and dried to obtain 3.8 kg seaweed meal.

Example 4: Preparation of Seaweed Meal from Dried Gracilaria

[0034] 10 kg dried Gracilaria was mixed with 150 kg water in the tank, and stirred at 60 RPM for 30 minutes to clean the seaweed. The water was removed to obtain 50 kg clean seaweed. The clean seaweed was mixed with 250 kg 20% sodium hydroxide solution in the reaction tank. The mixture was heated and kept at a temperature of 100°C, with stirring at 60 RPM for 12 hours. Then the seaweed was washed with 600 kg water. The water was removed to obtain 45 kg alkali pre-treated seaweed. The alkali pre-treated seaweed was mixed with 225 kg 0.5% nitric acid solution in the tank and stirred for 2 hours, and then washed with 100 kg water. The water was removed to obtain 42 kg acid pre-treated seaweed. The acid pre-treated seaweed was mixed with 210 kg 0.5% bleaching agent, sodium hypochlorous acid solution, stirred at 60 RPM for 120 minutes, and then the sodium hypochlorous acid solution was removed, followed by washing with 200 kg water. The water was removed to obtain 42 kg pre-treated seaweed. The specification of the pre-treated seaweed was: impurity of the red seaweed < 0.4%, the sulfate content of the seaweed < 0.1 %, and the color was light yellow.

[0035] The pre-treated seaweed was mixed with 210 kg of 300 U/g neutral cellulase solution, pH was adjusted to 8.0, heated and the temperature was kept at 70°C, with stirring at 60 RPM for 6 hours. The cellulase solution was removed and the seaweed was washed with 200 kg water. 32 kg cellulase treated seaweed was obtained.

[0036] The treated seaweed was mixed with 64 kg water and processed by HAAKE Process 11 Parallel Twin Screw Extruder with heating temperature set at 140°C. And then the extruded seaweed was crushed and dried to obtain 2.5 kg seaweed meal.

Example 5: Preparation of Seaweed Meal from Dried Gracilaria

[0037] 10 kg dried Gracilaria was mixed with 120 kg water in the tank, and stirred at 60 RPM for 30 minutes to clean the seaweed. The water was removed to obtain 50 kg clean seaweed. The clean seaweed was mixed with 150 kg 10% sodium hydroxide solution in the reaction tank. The mixture was heated and kept at a temperature of 90°C, with stirring at 60 RPM for 4 hours, then the seaweed was washed with 600 kg water. The water was removed to obtain 48 kg alkali pre-treated seaweed. The alkali pre-treated seaweed was mixed with 150 kg of 0.4% sulfuric acid solution in the tank and stirred for 1 hour, and washed with 100 kg water. The water was removed to obtain 48 kg acid pre-treated seaweed. The acid pre-treated seaweed was mixed with 150 kg of 0.3% bleaching agent, hypochlorous acid solution, stirred at 60 RPM for 1 hour. And then the sodium hypochlorous acid solution was removed, and the seaweed was washed with 100 kg water. The water was removed to obtain 46 kg pre-treated seaweed. The specification of the pre-treated seaweed was: impurity of the red seaweed < 0.6%, the sulfate content of the seaweed < 0.3%, and the color was light yellow.

[0038] The pre-treated seaweed was mixed with 150 kg of 200 U/g alkaline cellulase solution, pH was adjusted to 8.0, and heated and kept at 60°C, with stirring at 60 RPM for 4 hours. The cellulase solution was removed and the seaweed was washed with 100 kg water. 42 kg cellulase treated seaweed was obtained.

[0039] The treated seaweed was mixed with 42 kg water and processed by HAAKE Process 11 Parallel Twin Screw Extruder with heating temperature set at 120°C. And then the extruded seaweed was crushed and dried to obtain 3.5 kg seaweed meal.

Example 6: Preparation of Seaweed Meal from Dried Gracilaria

[0040] 10 kg dried Gracilaria was mixed with 150 kg water in the tank, and stirred at 60 RPM for 30 minutes to clean the seaweed. The water was removed to obtain 50 kg clean seaweed. The clean seaweed was mixed with 120 kg of 4% sodium hydroxide solution in the reaction tank. The mixture was heated and kept at a temperature of 85°C, with stirring at 60 RPM for 2 hours, and then washed with 600 kg water. The water was removed to obtain 49 kg alkali pre-treated seaweed. The alkali pre-treated seaweed was mixed with 150 kg of 0.4% sulfuric acid solution in the tank and stirred for

45 minutes, followed by washing with 100 kg water. The water was removed to obtain

46 kg acid pre-treated seaweed. The acid pre-treated seaweed was mixed with 150 kg of 0.3% bleaching agent, chlorine dioxide solution, stirred at 60 RPM for 1 hour. The chlorine dioxide solution was removed, and the seaweed was washed with 100 kg water. The water was removed to obtain 46 kg pre-treated seaweed. The specification of the pre-treated seaweed was: impurity of the red seaweed < 0.8%, the sulfate content of the seaweed < 0.4%, and the color was light brown. And then the seaweed was crushed and dried to obtain 4.2 kg seaweed meal.

Example 7: Preparation of Seaweed Meal from Ahnfeltia

[0041] 10 kg Ahnfeltia was mixed with 150 kg water in the tank, and stirred at 60

RPM for 30 minutes to clean the seaweed. The water was removed to obtain 49 kg clean seaweed. The clean seaweed was mixed with 120 kg of 5% sodium hydroxide solution in the reaction tank. The mixture was heated and kept at a temperature of 88°C, with stirring at 60 RPM for 120 minutes, and then washed with 500 kg water. The water was removed to obtain 47 kg alkali pre-treated seaweed. The alkali pre-treated seaweed with 120 kg of 0.39% oxalic acid solution in the tank and stirred for 30 minutes, followed by washing with 100 kg water. The water was removed to obtain 48 kg acid pre-treated seaweed. The acid pre-treated seaweed was mixed with 120 kg of 0.15% bleaching agent, hypochlorous acid solution, stirred at 60 RPM for 30 minutes. Then the hypochlorous acid solution was removed, and the seaweed was washed with 100 kg water. The water was removed to obtain 46 kg pre-treated seaweed. The specification of the pre-treated seaweed was: impurity of the red seaweed < 0.5%, the sulfate content of the seaweed < 0.3%, and the color was light yellow.

[0042] The pre-treated seaweed was mixed with 120 kg of 110 U/g acid cellulase solution, pH adjusted to 4.80 with hydrochloric acid, heated and the temperature was kept at 45°C, with stirring at 60 RPM for 180 minutes. The cellulase solution was removed and the seaweed was washed with 100 kg water. 42 kg cellulase treated seaweed was obtained.

[0043] The treated seaweed was processed by HAAKE Process 11 Parallel Twin Screw Extruder with heating temperature set at 130°C. And then the extruded seaweed was crushed and dried to obtain 3.6 kg seaweed meal. Example 8: Preparation of Seaweed Meal from Dried Gelidium

[0044] 10 kg dried Gelidium was mixed with 100 kg water in the tank, and stirred at 60 RPM for 30 minutes to clean the seaweed. The water was removed to obtain 50 kg clean seaweed. The clean seaweed was mixed with 100 kg of 5% sodium hydroxide solution in the reaction tank. The mixture was heated and kept at a temperature of 60°C, with stirring at 60 RPM for 6 hours, and then washed with 600 kg water. The water was removed to obtain 49 kg alkali pre-treated seaweed. The alkali pre-treated seaweed was mixed with 100 kg of 0.2% phosphate acid in the tank and stirred for 1 hour, followed by washing with 100 kg water. The water was removed to obtain 48 kg acid pre-treated seaweed. The acid pre-treated seaweed was mixed with 150 kg of 0.06% bleaching agent, sodium hypochlorous acid solution, and stirred at 60 RPM for 1 hour. The sodium hypochlorous acid solution was removed, and the seaweed was washed with 100 kg water. The water was removed to obtain 46 kg pre-treated seaweed. The specification of the pre-treated seaweed was: impurity of the red seaweed < 0.5%, the sulfate content of the seaweed < 0.4%, and the color was light brown.

[0045] The pre-treated seaweed was mixed with 150 kg of 80 U/g acid cellulase solution, pH was adjusted to 5.5, heated and the temperature was kept at 40°C, with stirring at 60 RPM for 1.5 hours. The cellulase solution was removed and the seaweed was washed with 100 kg water. 43 kg cellulase treated seaweed was obtained.

[0046] The treated seaweed was mixed with 43 kg water and processed by HAAKE Process 11 Parallel Twin Screw Extruder with heating temperature set at 115°C. And then the extruded seaweed was crushed and dried to obtain 3.2 kg seaweed meal.

Example 9: Preparation of Seaweed Meal from Dried Porphyra

[0047] 10 kg dried Porphyra was mixed with 150 kg water in the tank, and stirred at 60 RPM for 30 minutes to clean the seaweed. The water was removed to obtain 50 kg clean seaweed. The clean seaweed was mixed with 200 kg of 15% calcium hydroxide solution in the reaction tank, heated and kept at a temperature of 90°C, with stirring at 60 RPM for 6 hours, and then washed with 500 kg water. The water was removed to obtain 50 kg alkali pre-treated seaweed. The alkali pre-treated seaweed was mixed with 150 kg of 0.4% phosphate acid in the tank and stirred for 45 minutes, followed by washing with 150 kg water. The water was removed to obtain 49 kg acid pre-treated seaweed. The acid pre-treated seaweed was mixed with 150 kg of 0.25% bleaching agent, sodium hypochlorous acid solution, stirred at 60 RPM for 45 minutes, and then the sodium hypochlorous acid solution was removed, and washed with 100 kg water. The water was removed to obtain 48 kg pre-treated seaweed. The specification of the pre-treated seaweed was: impurity of the red seaweed < 0.4%, the sulfate content of the seaweed < 0.5%, and the color was light yellow.

[0048] The pre-treated seaweed was mixed with 150 kg of 220 U/g acid cellulase solution, pH was adjusted to 6.0, heated and the temperature was kept at 60°C, with stirring at 60 RPM for 1.5 hours. The cellulase solution was removed and the seaweed was washed with 100 kg water. 46 kg cellulase treated seaweed was obtained.

[0049] The treated seaweed was mixed with 46 kg water and processed by HAAKE Process 11 Parallel Twin Screw Extruder with heating temperature set at 135°C. And then the extruded seaweed was crushed and dried to obtain 3.8 kg seaweed meal.

Example 10: Preparation of Seaweed Meal from Dried Gracilaria

[0050] 10 kg dried Gracilaria was mixed with 150 kg water in the tank, and stirred at 60 RPM for 30 minutes to clean the seaweed. The water was removed to obtain 58 kg treated seaweed. The specification of the treated seaweed was: impurity of the red seaweed < 0.8%, and the color was light brown. And then the seaweed was crushed and dried to obtain 6.2 kg seaweed meal.

Example 11 : Basic Analysis of the Seaweed Meal Preparations

[0051] Table 2 summarizes the treatment of Examples 2-10 to obtain various seaweed meal preparations.

[0052] The gel strength measurement was performed as follows: 3.0 g of dry seaweed meal was mixed with 197 g of distilled water in a pre-weighed beaker. The mixture was heated to 90°C, with stirring slowly using a spoon, while placed in a 90°C water bath for 30 minutes until the seaweed meal was dissolved in the distilled water. The water jelly was allowed to cool for 3 hours at room temperature. The beaker containing the jelly was transferred to a 20°C constant temperature box and stored for at least 20 hours before the gel strength measurement. The gel strength was tested by TXAT2i (Texture Analyser).

[0053] The melting point and the gelling temperature measurement was performed as follows: 2 g of sample was mixed in 200 ml deionized water, the gelling temperature and melting point were measured by Anton Paar MCR 301 rheometer, with the following parameters:

1. Heating ramp: 2°C/min, 0.4 Hz, 1 %;

2. Stabilization: 15 min, 0.4 Hz, 1 %;

3. Cooling ramp: 2°C/min, 0.4 Hz, 1 %; and

4. Stabilization: 15 min, 0.4 Hz, 1 %.

[0054] The sulfate test was performed as follows: 100 mg sample was dissolved in 20 ml of water (with heating if necessary), and 3 ml of barium chloride test solution and 5 ml of hydrochloric acid were added. The test solution was diluted and filtered if a precipitate formed. The solution or the filtrate was boiled for 5 minutes. A white, crystalline precipitate appeared.

[0055] The soluble dietary fiber contents of the samples were determined by the AOAC official methods, see for example, AOAC 985.29 and AOAC 991.43. The protein contents of the samples were determined by the established Kjeldahl method, published as ISO 20483:2006.

[0056] Table 3 summarizes the results of the basic analysis of the seaweed meal preparation samples 1 -9, obtained from Examples 2-10, in comparison with the commercially available seaweed powder and agar.

[0057] Sample 10 is a commercially available Gracilaria seaweed powder obtained from Viet Delta Industrial Company Limited (which was produced by washing the seaweed with water, drying the seaweed and crushing the dry seaweed to a powder form), and Sample 11 is a commercially available agar obtained from Green-Fresh Group, both were used as controls in the analysis. Sample 9 produced by a regular process was also used as a control. Samples 2 and 6 produced the best results in various tests. Samples 1 , 3, 4, 5, 7 and 8 also produced acceptable results in various tests.

[0058] Figure 1 shows the results of a typical Fourier Transform Infrared

Spectrometer (FTIR) of Sample 2 in comparison with Sample 10, commercially obtained seaweed powder, and Sample 11 , commercially obtained agar. The spectral feature at approximately 890 cm-1 is mainly associated with the C-H bending at the anomeric carbon in b-galactose residues. The presence of a strong band in the FTIR spectra at 930 cm-1 is indicative of the occurrence of 3,6-anhydro-D-galactose. 1550-1510 cm-1 is protein N-H bending (amide bands II). Thus, FTIR demonstrates that the process disclosed herein produced a seaweed meal having a lower protein content and a higher content of 3,6-anhydro-D-galactose compared to the commercially available seaweed powder and agar.

Example 12: Use of Seaweed Meal Samples in Preparing Stirred-type Yogurt

[0059] The seaweed meal samples obtained as described above, Samples 1-4 and 6-9 were used in preparing stirred-type yogurt. The commercially available seaweed powder, Sample 10, and the commercially available agar, Sample 11 , were used as controls. The stirred-type yogurt was prepared by dry blending a seaweed meal sample or a control and sugar, dispersing the dry blend to milk at 50-60°C and stirring for 15 minutes, adding water to the target weight, homogenizing at 65-68°C at 180/50 bar, pasteurizing at 95°C for 5 minutes, cooling down to 43°C and adding bacterial culture for making yogurt, incubating until the pH arriving at 4.6, smoothing the mixture and subjecting to post-fermentation, and refrigerating at 4°C.

[0060] Table 4 lists the ingredients for making the yogurt.

[0061] The viscosity and pH of each yogurt product were measured immediately after production. The sensory test was also performed. Additionally, observation of the yogurt products was made after 10 days storage, and after 20 days storage, respectively. Table 5 details the test results.

[0062] All yogurt products did not have much difference in the final pH. The yogurt products made with seaweed meal Samples 2 and 6 were the best in viscosity, and close to the yogurt product made with Sample 11 , commercially available agar. The yogurt products made with seaweed meal samples 1 , 3, 4, 5, 7, and 8 were lower in viscosity compared with the yogurt product made with Sample 11 , but still accecpable. Nevertheless, the yogurt products made with Sample 9 and commercially available seaweed powder, Sample 10, had lower viscosity than the other yogurt products. Lower viscosity caused thin mouthfeel of yogurt. Also, the yogurt products made with Sample 9 and Sample 10 demonstrated worse water holding ability and worse taste such as astringent and sticky mouthfeel than the other yogurt products.

[0063] All yogurt products made with seaweed meal Samples 1 -8 and 11 were stable under the storage condition for up to 20 days and no water separation was observed after 10 days or 20 days of storage. Nevertheless, there was a 2 mm water separation in the yogurt products made with seaweed meal Samples 9 and 10.

Example 13: Use of Seaweed Meal Samples in Preparing Set-type Yogurt

[0064] The seaweed meal samples obtained as described above, Samples 1-4 and 6-9 were used in preparing set-type yogurt. The commercially available seaweed powder, Sample 10, and the commercially available agar, Sample 11 , were used as controls. The set-type yogurt was prepared by dry blending a seaweed meal sample or a control and sugar, dispersing the dry blend to milk at 50-60 °C and stirring for 15 minutes, adding water to the target weight, homogenizing at 65-68°C at 180/50 bar, pasteurizing at 95 °C for 5 minutes, cooling down to 43°C and filling into containers, adding bacterial culture for making yogurt to each container, incubating until the pH arriving at 4.4, subjecting to post-fermentation, and refrigerating at 4°C. The ingredients are the same as those listed in Table 4 above. [0065] The pH of each yogurt product was measured immediately after production. The sensory test and shear stress test by a HAAKE Viscostester were also performed. Additionally, observation of the yogurt products was made after 10 days storage, and after 20 days storage, respectively. Table 6 details the test results.

[0066] The gel shear stress test demonstrates the gel capability of the yogurt products made with the seaweed meal or control in the order from high to low: S11 > S2 > S6 > S3 > S8 > S1 > S7 > S4 > S5 > S10 > S9. The gel strength is presented by the peak value, meanwhile brittleness is presented by the time course to arrive at the peak value. All of the yogurt products had similar brittleness but different gel strength. The gel strength of S9 made with seaweed meal Sample 9 (sample made with regular process) and S10 made with seaweed meal Sample 10 (commercially available seaweed powder) was lower than other yogurt products.

[0067] Additionally, the yogurt products made with Sample 9 and commercially available seaweed powder, Sample 10, had an undesirable astringent and sticky mouthfeel, worse water holding ability than the other yogurt products.

[0068] All yogurt products made with seaweed meal Samples 1 -8 and 11 were stable under the storage condition for up to 20 days and no water separation was observed after 10 days or 20 days of storage. Nevertheless, there was a 2 mm and 3 mm water separation in the yogurt products made with seaweed meal Samples 9 and 10, respectively.