_T his invention refers to a process for the obtention of carho- 5 hydrates, derived from vegetal raw material, thus fitting in the technical sector of the industrial production of carbon hv drates.

In its essential form, the utilization of carbohydrates is widely spread all over the world; when fermented, the prcduc- 0 tion of alcohol would suffice to justify their existence.

In the sector of estarification the production of biodegrada¬ ble detergents, whose non polluting importance does not need to be enhanced would already represent a large sector. By acid degradation, .besides other compounds they produce Fur- fural; by the alkaline one, lactic acid with a range of appli¬ cations encompassing food, esters, pla≤ticizers and plastics. By oxidation and allylation and the very broad range ofEydroge nation of carbohydrates, with numberless secondary reaction food, resins, plastics, plasticizers, explosives, cosmetics, adhesives, etc. are obtained.

To restrain ourselves from further designations, beyond the scope of this patent, we would like to emphasize that any of the larger branches: Hydrogenation - Fermentation - Ξsterifica tion, by itself justify it. For centuries, the process of obtaining essential saccharoids has not changed. The equipment changed, improved becoming exceedingly sophisticated. Sophistication reached a stage that brought forth the dissapearance of small and medium size sugar plants, leaving only .the macro-units. This because the process was exclusively mechanic. The basic operation was to evaporate the water corresponding to the sugar cane juice used; no che_τ_:L cal reaction was introduced. Only sucrose was wanted, the purer the better. All products that were not sucrose were rejected. The other compounds, even if valid and usef l were discarded as waste, even if valuable. This way no major change was intro¬ duced in the production process of carbohydrates of vegetal nature in the last centuries. Only the mechanical processes were improved, with the coming of steam, vacuum, electricity, but the process remained the same. It was based on the crvstal

lization of sucrose and the simple separation of the solid frαn the liquid stage. This fact, base of the ancient process is of foremost significance for the understanding of the difference from the new process we shall proceed to describe. _A t the current stage, the large scale production of carbohy¬ drates, from sugar cane, beets, sorghum or any syrups able to produce crystalline carbohydrates, proceeds along the following overall line: initially, the raw-material passes in machines that mechanically remove the juice followed by thermic precipi- tation, passing to the vacuum concentration and crystallization, ending with the separa-tion of the solid _t and liquid stages. Practically, all the production of carbohydrates, the condensa¬ tion ones of high molecular weight, and those of three, five and six carbons, leads to the obtention of the best utilized soluble form: the al a-delta-glucopyranosis.

It is known that ^* the types of carbon hydrates most common in nature are cellulosis and starch, the last one as reserve nourishment for plants and the former constituing their skel¬ eton, both nonsoluble in water. With the exception of these two, the carbohydrates ocurring in nature form solutions part

• of which can be transformed in solid matter and part that - inspite of the technological progresses - remains liquid, and might reach high quantitative.levels.From the industrial point of view, the existence of the two stages is unfavorable because, besides the depletion of the solid phasis, the liquid one is subject to chemical and biological reaction, normally not controlled.

Inversely, in the new procεs-s specific controlled enzymatic reactions are introduced, only on previously determined sub— strates, without reacting with the remaining components, permitting to obtain, only the solid phasis. This is one of the main points of the process. The process for- the obtention of carbohydrates derived from vegetal raw-material is the subject of this Patent Application. With this process, by the judicious utilization of adequate hydrolases and of transferases of the enzymes classification, it is possible to obtain carbon hydrates totally crystalliza- ble and soluble that might be stored over long time periods without decomposition, permitting developments like hydrogen-

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ation, esterification, oxidation, fermentation, acid and alka¬ line degradation, propylenic reaction, etc.

In the process, the utilization of enzymes reacting in sucessive steps, allows the total transformation of solids in carbohy- drates of six to twelve carbons, for direct use or for further developments, arising from a primary product exclusively .in the solid stage. This way, the action of hydrolases-oxiredutases- transferases, acting stepwise, permits to reach specific and final sectors that sometimes even nature does not manage to attain.

The drafts included in this report, illustrate fully the intend¬ ed innovations, figure 1 shows the overall flow chart of the conventional procedures currently used in industrial chemistry, whereas figure 2 shows the flow chart of the actual process. In more detail, in figure 1 we observe an (1) expeller where the juice is extracted by any pressure procedure, by cylinder or diffusion; the filters (2) with a wire grid where the extracted juice is strained to remove the gross material in suspension; the heaters (3) where the juice is cleaned by heat, chemical coagulants and even ionic exchange; the plate heaters (4) to increase the extracts temperature; the vacuum concentrator batteries (5) where most of the water is eliminated; the so- lidifiers unit (6) where, after passing again through plate heaters (4) solids are separated from the liquid, forming a mixed paste, ending the process in the centrifugal separators (7) , where solids are -finally separated. Figure 2 shows that, in the current process, some of the heaters (3) the centrifugal separators (7) and some plate heaters (4) are excluded whereas the other conventional components remain. The proposed procedure acts on the extracts, creating controlled conditions that induce directions on the dynamics of the reac¬ tions, which are able to complete nature's action; it acts on pre-established substrates, enabling the obtention of a, specific and defined product that basically maintains all the initial components, as only the reactions .that were interrupted in the so called "relative vegetal cycle" are completed.

With the diffuser (1) the extract or juice is obtained were the ■ dissolved carbohydrates are found; there is no need to control the pH ^' , as the vegetal extracts are in the 3, 5 and 7 range were

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the proposed enzymes act and the reaction times vary Iran 20 to 90 in. Temperature should remain at 18C to 67C-To simplify the enzymes that should be added are listed below, by the num¬ ber corresponding to their international classification, and 5 in the present case they vary from 3.2.1.1(4-glucan-4-glucanhv drolase) to 5.3.1.5 (xylose-ketol-isomerase) ; during the reac¬ tions:

Name Quantity

1st. step with addition

10 3.2.1.4-3-1,4 Glucanhydrolase ^" l,5ϋ

3.2.1.15 -<* -1, -1 Galacturonic Glucanhydrolase 0,9U 2nd step with addition

3.2.1.20- ""-L-D Glucoside glucohydrolase 0,3U

3rd step with addition

15 3.2.1.21 - B-D Glucoside glucohydrolase 0,3U

5.3.1.4 - D Arabinose-ketol-isomerase 0,15U

5.3.1.5 - D Xylose-ketol-isomerase 0,15ϋ 4th step with addition 3.2.1.1 -<— 1,4 Glucan-4-glucanhydrqlase 0,12U

20 3.2.1.3 -- ^■ -- 1,4 Glucohydrolase 0,22U is reached.

The enzyme unit is the quantity able to hydrolize in 10 in. one mαl of starch. Once the initial stage is ended the temperature in the heat-

25 ers (3) is raised -to 90/100C to clot all impurity that re¬ mains and the albuminoids, then proceeding to a filtration or cεntrifugation to obtain the clean syrup around 13<?3rix; the syrup is led to the vacuum concentrators (5) to eliminate practically all water, at a temperature not beyond 70C around

30 709Brix. Once the adequate saturation is reached to form the crystals, the product is mixed at low speed in the crystal- lizers (β) till the granules separate by themselves. The end product must have a relative humidity of less than 0,50%, preferrably at 0,25%. The obtained concentrate is com-

35 posed of carbohydrates without liquid stage. ^' The process permits, as an option, that,the concentrate be sent to a spray-dryer to form an even powder or granulate to be used directly or for further industria "se in types of reactions mentioned previously.

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Among the process' advantages one might mention:

1) no losses in the primary process;

2) a sole solid phasis in the prepared primary product;

3) strictly natural product composition;

4) absence of effluents and by-products;

5) about 30% higher end production;

6) greater storage and transportation facility;

7) lower production cost because of the low enzyme quantities employed;

8) elimination of part of the conventional equipment currently used;

9) process' simplicity with consequent industrial costs reduc¬ tion; and,

10) integral solid stage for natural components, without de¬ pletion due to partition of stages. Below a comparative plan of the two processes: Conventional Enzymatic a) Extraction a) Extraction b) Chemical precipitation b) - c) - ^• c) Enzymatic treatment d) Filtration - d) Filtration e) Evaporation e) Evaporation f) Cooking f) Cooking g) Crystallization g) Crystallization

(final) h) Centrifugation (triple) Integral Sugar

Sugar (sucrose) syrup (discard)

Note that, in the Enzymatic process, the operation for the obtention of a dry crystalline product, composed of a sole solid phasis ends in the crystallize . In the Conventional this stage is constituted by the so called "baked paste" a mixture of crystals and honey, that must be separated, if not both will be lost by fermentation and further reactions. Thus the following conclusions might be reached for the enzymatic process: a) Sugar and all other solid components found in the initial juice are present. b Absence of secondary carbohydrates, like gums, colloids,

xylose, arabinoses, starch, dextrines, etc., that normally are agents for the high viscosity of syrups. To decrease this viscosity the use of surfactants or detergents is re¬ quired. In this process all secondary carbohydrates are transformed in glucose. c) Reduction of darkening agents and pigments like Antocyanins and Polyphenols (unwanted because of color and taste) . d) Reduction of operational costs and elimination of equipment. e) Increse of final revenues. f) Obtention of natural carbohydrate without partion between solid and liquid stages and fully able of further develop¬ ments. Aiming to further individualize the differences and advantages of the enzymatic process, we present a summary of the average composition of final components in the two processes:

Convent!onal Enzymatic

(all solid)

Sugar -

Sucrose: 99, 5% Treacle JSucrose 80%

Sucrose: 35% Main a)

Glucose: 7% ► carbohy¬ 1.Glucose 10%

Fructose: 9% drates

Galactose

Xylose

Ramnose

Arabinose

Pectins Starch

D. Manitol ϋronic ACJ -d Methoxyl

Conventional (cont.) Enzymatic (cont. )

Dextrines Secondary ^' so ₃ 0,71% carbohy-

Gums Cl 0,71% drayes d> -

Colloids (liquid) ^P 2°5 0,05%

Insolubles 0,24%

Aconit c Acid other carbohy¬ e) Proteins 1,59%

Inosite drates

Phytin (liquid) f) Aminoacids g) Others nitrogenous compound 0,85% h) Aconitic acid 1,19% i) No nitrogenous acid 0,85% j) Esteroids-Waxes-Present 1) Pigments - Present

SO., 1,8% m) Vitamins 3 complex

Cl 1,8% Acid Ashes ₂ A ₅ 0,06% (liquid)

Insolubes 0,20%

Total ashes: 12% (liquid) Proteins: 4,5% (liquid) Aminoacids (liquid) Other nitrogenous compounds - 2,0%

Esteroids-Waxes

-Antocyanins

Pigments

-Cloriphyll (liquid)

-Polyphenols

-Tannin

Vitamins B Complex

Water - 17/25% simple comparaison shows the large number of compounds that were transformed in glucose and from inactive they become the most active carbohydrate, D-Glucopyranose. Furthermore they were not destroyed but reshu led by the enzymes that can lead them to a more suitable and usable form.

This reactions sequence that would be obtained in nature byits "relative cycle" was achieved by enzymatic action. It should be noted that in the conventional product only sucrose is found as a solid, and this is an important fact. The results derived in assays of alcoholic fermentation clearly show the advantage of ^* the product obtained by enzymatic action ^, An assay was undertaken for alcoholic fermentation obtaining etanol by the following process: a) Must preparation dissolving 140g of carbohydrate in water ^' to make up a liter. b) Adjustment of _. the pH at 4,7 by addition of sulphuric acid. c) Innoculation with 50g of pressed saccharomyces cerevisiae per liter of must. d) Maintenance of the required fermentation conditions (1 liter cylinder at 30-32C) . Fermentation reached completion in 6/7 hours. e) Output: in well controlled technical conditions output might reach 90%.

■ The results obtained by assays of ^* alcoholic fermentations clearly show the advantage of using carbohydrate, obtained by enzymatic action, over the conventional raw-material: - In the production of alcohol the used syrup contains 55 to ^■ 60% of Total Reducing Sugars. The carbohydrate of the ^' enzy¬ matic route has 90 to 92% of TRS. - The syrup exhibits some non fer entescible carbohydrates. In this enzymatic process all carbohydrates are fermentescible. There is an 82% output without addition of nutrients, reaching 90/92% when nutrients are added, better results than those of the syrup fermentation. The end product (being, integral sugar) , might, further be a - source of carbon interesting for the production of yeast due to the advantages of easy storage, transport and handling. Experiences were undertaken with integral pure sugar as well as with sugar enriched with salts and the results showed high yelds in the production of the yeast in less time than with the conventional processes.

The end product obtained by the current process has a range of applications; as human nourishment because it possesses besides

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high rates of sugars, around 92%, mineral salts, essential and non essential amino-acids, vitamins, etc. as nourishment for animals and for industrial scopes it is efficiently used as raw material for the process of alcoholic, glutamic, acetic, fermentation and also the production of a large range of prod¬ ucts.

The process, object of the current invention, has, among others the following advantages: a) reduces industrial costs (30% in machinery and equipment) b) increases production attaining integral sugar, 50% per ton of sugar cane with Pol 88, as it does not use chemicals. Further with this enzymatic process a natural sugar is obtained totally in solid state, in a sole phasis, without wastes or secondary carbohydrates. Therefore, the following application for patent, presents a great innovation and the product(s) obtained possess by far superior characteristics to the conventional ones.