CN107006673A - Protein product and preparation method thereof - Google Patents

Abstract

The invention provides a kind of protein product, the protein product includes alcohol soluble protein and carbohydrate, it is characterised in that the alcohol soluble protein accounts for more than the 70wt% of albumen butt；Meanwhile, alpha-alcohol soluble protein accounts for more than the 75wt% of the alcohol soluble protein, and the molten albumen of β -ol accounts for below the 20wt% of the alcohol soluble protein, and the molten albumen of γ -ol accounts for below the 6wt% of the alcohol soluble protein.

Description

Protein product and preparation method thereof

Technical field

The present invention relates to the method that the method for alpha-alcohol soluble protein and purification obtain protein product is enriched with from raw material, also relate to And the protein product obtained by the above method, belong to crops manufacture field.

Background technology

Zeins (zein, zein) be it is a kind of from corn or the material containing zein, such as maize yellow-powder raise The production extracted in material (also known as corn protein powder, corn gluten meal) or the product such as corn distiller's dried grain and DDGS (DDGS) Product.Alcohol soluble protein is content highest albumen, the 44wt%~79wt% accounted in corn embryosperm albumen in corn embryosperm.Maize Powder is mainly obtained from cornstarch wet production.In corn wet production, sulfurous acid soaking corn, soaking water are prepared first Carry out evaporating and obtaining corn steep liquor after separating with corn, carried out after corn while it is primary and secondary it is broken apart go out plumule, then endosperm Fiber is isolated by fine grinding, and starch milk in main separating step is separated seitan after pre-concentration, starch warp Cross after washing by dehydration, the finished product dried；And maize yellow-powder is made after concentrating, being dehydrated, dry in above-mentioned seitan；It is above-mentioned Plumule obtains the plumule dregs of rice after being dehydrated, drying and extract corn oil, and above-mentioned fiber is distinguished after dehydrating with the plumule dregs of rice The products such as fibrin feed, high dregs of rice feed are obtained after spray corn steep liquor.

Zeins conduct《Chinese Pharmacopoeia (2010)》In pharmaceutic adjuvant, also obtain U.S. FDA approval, be public Recognize the material being directly added into food of safety (GRAS).Corn protein has the film forming characteristics of surface-active and uniqueness, Bulk support, food fresh keeping film, biodegradable packaging material, chewing gum, azelon etc. can be targetted as medicament, in food The application of the fields such as product, medicine, chemical industry is increasingly extensive.

Zeins belongs to protein,alcohol-soluble matter, the aqueous solution of ethanol, isopropanol or acetone can be dissolved in, in pure water Or it is insoluble in absolute ethyl alcohol.It is main comprising four kinds of components, i.e., alpha-alcohol soluble protein (account for the 70wt% of zeins~ 85wt%), the molten albumen of β -ol (accounting for 1wt%~5wt%), the molten albumen of γ -ol (accounting for 10wt%~20wt%) and δ -ol are molten Albumen (accounting for 1wt%~5wt%) (Cereal Chemistry, 2011,88 (2):159-173).Wherein, alpha-alcohol soluble protein is Topmost commercial corn alcohol soluble protein.Processing is prepared zeins technique and mainly existed using different Gliadin Components Difference in solubility in Extraction solvent.The maize yellow-powder of the above-mentioned output in corn wet milling process, dry protein content is about 65wt% or so, the also main starch, 10wt%~13wt% fiber and about 6wt% for including about 15wt%~20wt% Fat, is also one of primary raw material that commercial corn alcohol soluble protein is extracted.Commercial process mainly uses Swallen1941 Patent (US2287649A) and Carter and the Reck patents of 1970 (US3535305), as shown in Figure 3.With isopropanol or 95% ethanol (v/v) handles maize yellow-powder in batches or continuously in higher pH value and temperature (50-60 DEG C), extract filtering or from The heart, then precipitate alcohol soluble proteins, the vacuum dried finished product that is milled to obtain with (- 10 DEG C~-25 DEG C) excessive of cold water or low temperature.Certainly, Prepare zeins and also have other or modified technique, but be all to use organic solvent (US6602985B1；Cereal Chemistry Journal,2006,83(5):565-568；Cereal Chemistry,2011,88(4):356-362) or have Machine acid (Cereal Chemistry, 2008,85 (2):It is 202-206) process program of extractant.

Also there is researcher to carry out research removing to the non-protein impurity of protein raw materials, improve corn total protein content, or by This improves the efficiency of follow-up solvent extraction.Cai Muyi et al. discloses one kind in patent CN101390564 and uses alkali heat-treatment Maize yellow-powder is so that partial starch, fat and pigment are changed into solvable so as to remove, so as to obtain with alcohol soluble protein and paddy egg Bai Weizhu protein isolate product.But due to there is the starch being connected with fiber or albumen in bloom, and pigment is (mainly beautiful Cream-coloured matter) active site of protein is embedded in, so it is difficult to being removed by single gelatinization or saponification.CPC International Inc. of the U.S. and Japan Showa Sangyo Co., Ltd. decolourizes to produce White corn protein (JP2004059537) using freezing, but the technique is related to methanol, third Ketone etc. has the use of the reagent of potential safety hazard.Sessa is a kind of disclosed in patent US20080242842 to use zeolite and activated carbon The method that the alcohol soluble protein for being dissolved in alcoholic solution is decolourized and is deodorized.In addition, also someone uses n-hexane or acetic acid second Ester equal solvent carries out decoloration and deodorization to alcohol soluble protein low moisture raw material or finished product.In addition, passing through the enzymes such as solvent or lipase Removing fat therein is so as to further improve total protein purity；And pass through chemical reagent such as ozone, persulfuric acid or peroxidating Thing or enzyme (such as lipoxygenase) decolourize.But, too high heating-up temperature also results in the amino acid in albumen Residue participates in some harmful substances of generation, and such as when temperature is more than 100 DEG C, cysteine and methionine can be with glucose response shapes Into noxious material --- acrylamide (Food Chemistry, 4th revised and extended edition, H.- D.Belitz, W.Grosch and P.Schieberle write, the 25-29 pages).Liaw et al. (US5968585A) utilizes UF membrane Technology separates the starch in endosperm and albumen, obtains feed liquid of the dry protein content in about 70wt%.Prepare corn alcohol at present It is high, molten still to there is solvent loss, use cost in the organic solvent of molten albumen or the various extraction processes of aqueous solutions of organic solvent The problem of agent wastewater treatment difficulty is big, significantly heating and cooling operation energy consumption is big is, it is necessary to further be solved.In addition, extracting The gelation that Cheng Zhonghui occurs is another drawback of organic solvent or aqueous solutions of organic solvent extraction and separation process, mainly due to The presence of the molten albumen of γ -ol and cause (Journal of Agricultural and Food Chemistry 60 (7): 1742-1747).There is process to be incubated 30min at 70 DEG C to prevent gel by pH regulations to 11.5, but albumen is in highly basic Deamidation or the destruction of sulfydryl of part peptide bond hydrolysis and asparagine and glutamine can occur in environment, in addition rear Phase acid adjustment step can also cause system to produce more salinity.Also research extracts alcohol using 90% acetic acid-aqueous solution (v/v) molten Albumen, but fat content therein is higher than alcohol extracting protein product, and it is used as relatively low (the Selling G of tensile strength of material W,Woods K K.Improved Isolation of Zein from Corn Gluten Meal Using Acetic Acid and Isolate Characterization as Solvent[J].Cereal Chemistry,2008,85(2): 202-206).Also technology is modified as the product of aqueous application by the alcohol soluble protein being chemically treated alcohol extracting (CN103781796A).But it is being entirely free of organic solvent (such as ethanol, isopropanol) or high content is organic currently without one kind The process of alcohol soluble protein is produced in the aqueous phase system of sour (such as 90% acetic acid).

The content of the invention

A kind of novel production process of alcohol soluble protein is found, while more effectively utilizing the other components money in related raw material Source turns into problem of the pendulum in face of those skilled in the art, but is also likely to become reduction production cost effective way the most.Therefore, A kind of separation under the conditions of the gentle acid-base value (pH=3~11) of organic solvent-free from the raw material comprising protein,alcohol-soluble is provided The alcohol soluble protein product gone out will be highly beneficial.The method that offer produces the product also will be highly beneficial.There is provided and implement The system of this method also will be highly beneficial.

The present inventor passes through to being found first after each component in zein raw material, including the research of different protein components： For wherein different component, especially protein component, corresponding enzyme and separation method can be used, organic solvent is not being included completely Aqueous environment (have that to can reach purifying corn alcohol soluble protein, especially α -ol in gentle acid-base value, pH=3~11) molten The effect of albumen, and the content in the product of β -/molten albumen of γ -ol can be controlled；Form a variety of useful by-products simultaneously Product, excellent opportunity is brought so as to be utilized to starch or alcohol production enterprise to the deep development of zein.

The first aspect of the present invention is related to a kind of method that alpha-alcohol soluble protein is enriched with from raw material, and the raw material is molten comprising alcohol Albumen and non-alcohol soluble protein, and optionally comprising macromolecular carbohydrate, (macromolecular carbohydrate in the present invention is main Including starch and cellulose) and/or grease, it is characterised in that methods described comprises the steps without using organic solvent：

(1) raw material is crushed and sized mixing；

(2) Protease Treatment is used, the molten albumen of at least a portion β -ol in raw material, the molten albumen of γ -ol and non-alcohol is molten Albumen carries out complete hydrolysis or partial hydrolysis, and removes hydrolysate using the filtering of particle diameter difference, so as to obtain alpha-alcohol soluble protein It is able to the crude product being enriched with；

(3) crude product is washed, be dehydrated, dried, obtain final products.

The second aspect of the present invention be related to it is a kind of from raw material purify obtain protein product method, the raw material comprising β- Alcohol soluble protein, the molten albumen of γ -ol and non-alcohol soluble protein, and optionally include macromolecular carbohydrate and/or grease, its feature It is, methods described comprises the steps without using organic solvent：

(1) raw material is crushed and sized mixing；

(2) using hydrolysis ferment treatment, in raw material at least a portion macromolecular carbohydrate carry out complete hydrolysis or Partial hydrolysis, and hydrolysate is removed using the filtering of particle diameter difference, so as to obtain albumen crude product；

(3) the albumen crude product is washed, be dehydrated, dried, obtain final protein product.

The third aspect of the present invention is related to the protein product according to made from aforementioned aspect of the present invention, the protein product bag Containing alcohol soluble protein and carbohydrate, it is characterised in that the alcohol soluble protein accounts for more than the 70wt% of albumen butt；Meanwhile, α- Alcohol soluble protein accounts for more than the 75wt% of the alcohol soluble protein, and the molten albumen of β -ol accounts for below the 20wt% of the alcohol soluble protein, γ -ol Molten albumen accounts for below the 6wt% of the alcohol soluble protein.

Above-mentioned aspect of the invention preferred embodiment in, raw material be selected from by maize yellow-powder, corn embryosperm karusen The group constituted with vinasse.

Above-mentioned aspect of the invention preferred embodiment in, protease be selected from by carboxyl protease, serine egg One or more in the group that white enzyme, metalloproteinases, thiol protease are constituted.Preferably, the carboxyl protease is mould Bacterium carboxyl protease, preferably aspergillus carboxyl protease, more preferably aspergillus oryzae carboxyl endo protease；The serine protease is The serine endoprotease of the serine protease of bacillus, preferably hay bacillus；The metalloproteinases be mould or The metalloproteinases of bacillus, preferably aspergillus oryzae metalloendoprotease or bacillus subtilis metalloendoprotease；Institute It is the thiol protease from plant, preferably bromelain and/or papain to state thiol protease.

Above-mentioned aspect of the invention preferred embodiment in, hydrolase be selected from by alpha amylase, carbohydrase, cellulose In the group that enzyme, 1,4 beta-glucanase, Pullulanase, zytase, pectase, arabanase, hemicellulase are constituted It is one or more.Preferably, the alpha amylase is the alpha amylase of mould or bacterium, preferably mould alpha amylase, more preferably aspergillus Alpha amylase；The carbohydrase is mould glucoamylase, preferably aspergillus or trichoderma glucoamylase；The cellulase For fungal cellulases, preferably trichoderma cellulase；The 1,4 beta-glucanase is fungi or bacterium 1,4 beta-glucanase；The general Shandong Blue enzyme is bacillus Pullulanase.

Above-mentioned aspect of the invention preferred embodiment in, when carrying out the Protease Treatment or hydrolysis ferment treatment, It is optionally added reagent composition enzyme is adjusted, the reagent composition is in the group being made up of following material It is one or more：The compound of the disulfide bond in albumen, such as phosphorus-containing compound or sulfur-containing compound can be opened, wherein phosphorous Compound preferably three (2- carboxyethyls) phosphine, compound of the sulfur-containing compound preferably containing free sulfhydryl group and/or can provide inferior sulfate radical Compound, more preferably mercaptoethanol, dithiothreitol (DTT), cysteine and oligopeptides comprising cysteine are (by 2-10 amino Acid composition peptide), sulphite, sulfurous acid, bisulfites, pyrosulfite；Metal ion, preferred as alkali ion, alkali Earthmetal cations, divalent transition metal ion, more preferably sodium ion, potassium ion, magnesium ion, calcium ion, manganese ion, cobalt ions, Zinc ion；Metal-chelator, preferably EDTA, EGTA.

Above-mentioned aspect of the invention preferred embodiment in, Protease Treatment is carried out under the following conditions：PH 3.5~ 10.5, preferably 3.8~10；20 DEG C~65 DEG C, preferably 35 DEG C~55 DEG C for the treatment of temperature；Processing time 0.2h~10h, preferably 0.5h ~5h.It is highly preferred that the condition is selected from：pH4.8、45℃；pH7.5、52℃；pH3.8、35℃；pH8.3、52℃； pH8.5、65℃；pH6.5、45℃；pH8.0、45℃；pH10.2、45℃；pH4.2、35℃；pH6.5、45℃；pH4.8、55 ℃；pH4.8、53℃；pH7.2、53℃；pH7.5、25℃；pH10.1、55℃.

Above-mentioned aspect of the invention preferred embodiment in, hydrolysis ferment treatment carry out under the following conditions：PH 3~8, It is preferred that 3.3~7.5, more preferably 4~6.5, most preferably 4.5~5.5；30 DEG C~72 DEG C for the treatment of temperature, preferably 35 DEG C~63 DEG C, more It is preferred that 40 DEG C~60 DEG C, most preferably 45 DEG C~55 DEG C；Processing time 0.5h~12h, more preferably preferably 1h~10h, 2h~8h, most It is preferred that 2h~7h；It is highly preferred that above-mentioned hydrolase treatment conditions are selected from：pH5.0、63℃；pH5.5、50℃；pH3.0、35 ℃；pH6.5、45℃；pH4.0、40℃；pH6.5、45℃；pH4.5、60℃；pH8、45℃；pH5、55℃；pH7.5、50℃； pH3.5、30℃；pH5.6、50℃.

Above-mentioned aspect of the invention preferred embodiment in, filtering is using 1 μm~80 μm, preferably 10 μm~50 μm of mistake Filter opening footpath is carried out, or is carried out using the membrane filtration aperture of 10nm~10 μm, preferably 20nm~1 μm.

Above-mentioned aspect of the invention preferred embodiment in, alcohol soluble protein account for the albumen butt more than 74wt%, It is preferred that more than 80wt%, more preferably more than 85wt%, further preferred more than 90wt%, even more preferably more than 95wt%, Most preferably more than 97wt%, alpha-alcohol soluble protein accounts for more than the 77wt%, preferably more than 85wt%, more preferably of the alcohol soluble protein More than 90wt%, most preferably further preferred more than 95wt%, 100wt%, the molten albumen of β -ol account for the alcohol soluble protein Below 10wt%, preferably below 5wt%, more preferably below 3wt%, further preferred below 2wt%, most preferably 0%, γ -ol Molten albumen accounts for below the 10wt%, preferably below 5wt%, more preferably below 2wt% of the alcohol soluble protein, most preferably 0%, albumen Butt account for more than the 85wt%, preferably more than 90wt%, more preferably more than 94wt%, most preferably 99wt% of the protein product with On.

Compared with method prepared by prior art alcohol soluble protein, the technical scheme involved by the application has the following advantages that：

Total protein content and alpha-alcohol soluble protein, the molten albumen of β -ol in alcohol soluble protein product of the present invention, γ -ol The content of molten albumen is with using other organic solvents such as ethanol, isopropanol according to traditional extraction process products obtained therefrom without substantially poor Not.Alpha-alcohol soluble protein is to provide functional main protein component such as product film forming.Alcohol soluble protein production of the present invention The product that content of the alpha-alcohol soluble protein in total protein in product can be prepared higher than traditional handicraft.

In the case where that need not carry out single decoloration process, the color and luster of alcohol soluble protein product of the present invention is than tradition The color of yellow alcohol soluble protein product prepared by method is more shallow, and accessible white, also has than product made from conventional method Lower corn characteristic odor, the wider and single food samples consumption of the application of product can be bigger without influenceing former food Apparent and local flavor.

Alcohol soluble protein product of the present invention is with using other organic solvents such as ethanol, isopropanol according to traditional extraction side Method products obtained therefrom such as alcoholic solution dissolubility, film forming, into fibroid, mouldability, prepare the functions such as microballoon characteristic, degradability Without significant difference in terms of characteristic, some characteristics, such as microballoon aqueous stability are even better than product prepared by traditional handicraft.

The preparation method of alcohol soluble protein product of the present invention can be realized with a step retains alpha-alcohol soluble protein in selectivity While any regulation β -ol molten albumen, the content of the molten albumen of γ -ol in the product, without according in traditional preparation methods Using the aqueous solution of organic solvent, and alpha-alcohol soluble protein, the molten egg of β -ol are separated by adjusting the concentration of organic solvent step by step The white and molten albumen of γ -ol.

The preparation method of alcohol soluble protein product of the present invention, due to being carried out in whole water phase system, even if reservation γ- Alcohol soluble protein can also avoid the generation of the material gelation in preparation process completely, you can output remains with the molten albumen of γ -ol Product, so as to widen the type of alcohol soluble protein product significantly.

The preparation method of alcohol soluble protein product of the present invention, in starch minuent degraded (DE values<Or the non-paste of starch 40) Under conditions of change state, you can realize the separation of starch and its spin-off and alcohol soluble protein.

The preparation method of alcohol soluble protein product of the present invention in output alcohol soluble protein product simultaneously, can coproduction raw sugar production Product or oligosaccharides, disaccharides, the refined material of monose product, other feed grades or food-grade albumen class byproduct.

The preparation method of alcohol soluble protein product of the present invention compares traditional preparation methods, without special decolouring, deodorization Technique can obtain bright color, milky white, the flat alcohol soluble protein product of original characteristic odor.

For the process involved by the preparation method of alcohol soluble protein product of the present invention, because whole process only makes It is medium with water, and belongs to fire without using the ethanol used in traditional extraction technique, acetone, n-hexane, ethyl acetate etc. The solvent of dangerous Class A, therefore workshop explosive-proof grade is relatively low, production process security is good.

Equipment involved by the preparation method of alcohol soluble protein product of the present invention is chemical process, food and medicine is processed Common equipment, equipment investment cost is relatively low.Also, raw materials used need not dehydrate can be prepared, it is possible to decrease part Energy consumption.

Brief description of the drawings

Fig. 1 is the process flow diagram of alcohol soluble protein production technology of the present invention.

Fig. 2A and 2B are the production equipment schematic diagrames of implementation alcohol soluble protein production technology of the present invention.

Fig. 3 is the main process of zeins of the prior art.

Fig. 4 A-4E are carrying out reproducibility dodecane for the protein component in the raw material and different zein products of example After base sodium sulphate (SDS) polyacrylamide gel electrophoresis electrophoretogram (under) and OD value analysis chart (on).Wherein, A is original Material；B is the alpha-alcohol soluble protein product that ethanol extracts (conventional method)；C is the alpha-alcohol soluble protein product that this patent method is obtained；D It is the alcohol soluble protein composition product that this patent method is obtained with E.

Fig. 5 is that zeins is embodiment 1-6 and film made from the zeins obtained by comparative example is broken Stress sheet.

Embodiment

Zein provided by the present invention and preparation method thereof is described in further detail below in conjunction with the accompanying drawings.

Fig. 1 is the process flow diagram that alcohol soluble protein is extracted from zeins raw material.Raw material 102 is added water tune Defibrination 106 is carried out after section 104, sizeable material particles are obtained by 108, slurries containing the material particles enter respectively by In the enzymolysis piece-rate system 110 of enzymatic hydrolysis system 112 and piece-rate system 114 by series, parallel or connection in series-parallel composition, in 110 The a part such as carbohydrate of starch, fiber etc. and degraded or/and the molten protein component of modification non-alcohol in degradation material are simultaneously Component is separated.The isolate 116 rich in alcohol soluble protein is obtained by 110 and enters washing system 124, will then obtain being rich in alcohol The isolate 126 of molten albumen obtains alcohol soluble protein product after being dried 128.It can also obtain being rich in Protein Separation thing by 110 118, enter drying system 132 into drying system 132 or after carrying out refined 130, obtain protide product.May be used also by 110 Obtain being rich in carbohydrate isolate 120, can be utilized into carbohydrate recovery system 132.Digest piece-rate system The washings that the waste water and washing system of 110 outputs are produced enter water treatment system 136.

Zeins production technology equipment used in implementation process carries out exemplary explanation in fig. 2.Base This identical equipment be used for different raw material (such as maize yellow-powder, corn embryosperm karusen, distiller's dried grain) and purpose thing (comprising β-with The molten albumen of γ -ol and the alcohol soluble protein rich in alpha-alcohol soluble protein, the molten egg of alcohol comprising the molten albumen of β -ol and rich in alpha-alcohol soluble protein In vain, alpha-alcohol soluble protein).According to an exemplary embodiment, raw material used is maize slurry.According to another implementation Mode, raw material used can be corn embryosperm karusen dry powder.

Fig. 2A is the process flow diagram that alcohol soluble protein is extracted from maize slurry.According to an exemplary implementation Mode, maize slurry 202 is added water and entered after regulation tank 204 is mixed in pulverizer 206, for example, colloid mill can be used.

According to an exemplary embodiment, by the material moisture regulation in 204 to 95%；According to a preferred reality Mode is applied, by moisture regulation to 75%~90%.

As illustrated, feed liquid by after filter 208 with enzyme (such as complex cellulase), reagent composition (such as hydroxide Sodium, sodium pyrosulfite), water, steam etc. enter the first enzymolysis reactor 210.The He of the first separator 212 is sequentially entered after finishing 214, such as cyclone separator, material and enzyme (such as protease), reagent composition (such as sodium hydroxide, divalence of gained alcohol soluble protein Manganese ion, disodium ethylene diamine tetraacetate (EDTA-2Na)), water, steam etc. reacted into the second enzymolysis reactor 216, it is and laggard Enter centrifuge 218, such as three-phase disk centrifugal separator.

According to an exemplary embodiment, the material in 216 adjusts pH to 7.5 using sodium hydroxide, and addition accounts for egg Bai Chongliang 0.3% bacillus subtilis neutral proteinase and 0.5% bacillus alkaline protease, 0.1mM Mn²⁺At 55 DEG C Addition 0.05mM EDTA-2Na after carrying out 1.5 hours.

As illustrated, the material of the gained alcohol soluble protein of centrifuge 218 and enzyme (such as amylase), reagent composition (such as salt Acid), water, steam etc. enter the 3rd enzymolysis reactor 220.Filter 222 and 224 is sequentially entered after completion of the reaction, and 224 After middle realization washing alcohol soluble protein finished product is obtained into drier 228.

According to exemplary embodiment, device 222 (for example can be micro-filtration film device) and 224 (for example can be plate Frame filter press) final separation of the molten protein component of non-alcohol and alcohol soluble protein can be achieved.

As illustrated, the protein component of the non-alcohol soluble protein of the gained of centrifuge 218 can be with enzyme (such as protease), reagent set Compound (such as hydrochloric acid), water, steam enter enzymolysis reactor 230 further processing after by filter bank 232 (such as 100nm~ 20nm milipore filters group) select after passing through and concentrating, it is dry that albumen is produced after being refined by chromatographic column 234 (such as ion exchange column) Product.212nd, 214 carbohydrate containing feed liquids may be recovered；218 third phase feed liquid and 224 washings can enter water process System.

Fig. 2 B are the process flow diagram that high-purity alpha-alcohol soluble protein is extracted from corn embryosperm karusen dry powder.According to One exemplary embodiment, after maize yellow-powder is crushed by pulverizer 240 (such as roll squeezer), adds into regulation tank 242 Water is mixed, and is squeezed into fiberizer 244, by entering after filter 246 in reactor 248.

According to exemplary embodiment, filter 246 can have the pore size filter of such as 200 mesh.

As illustrated, feed liquid and enzyme (such as compound cellulose/amylase), reagent composition (such as potassium hydroxide), water, steam Squeezed into after completion of the reaction in filter 250 Deng into first reactor 248.

According to exemplary embodiment, filter 250 can have the pore size filter of such as 300 mesh.

Separated as illustrated, being entered by 250 material in seperator 252 (such as horizontal screw centrifuge), gained contains The heavy phase material of alcohol soluble protein and enzyme (such as protease), reagent composition (such as sodium hydroxide, divalent calcium ions), water, steam Reacted into the second enzymolysis reactor 254.The trapped substance of filter 250 and the clear liquid of seperator 252 can carry out carbon hydrate Thing is reclaimed.

The enzyme reactor group (254-1 and 254-2) of two series connection is can also be according to exemplary embodiment, 254, Material is using salt acid for adjusting pH to 4.2 in 254-1 reactors, and addition accounts for telephoric acid neutral proteinase, the egg of protein by weight 0.8% Bai Chongliang 1% three (2- carboxyethyls) phosphines, are carried out 2 hours at 45 DEG C.Then adjusted into 254-1 reactors using potassium hydroxide PH to 8.3 is saved, addition accounts for the bacillus alkaline protease and 0.1mM Ca of protein by weight 0.3%²⁺0.5 is carried out at 50 DEG C small Shi Fanying.

As illustrated, 254 materials got are entered and separated in filter 256, retention material enters filter 258, washed (260) and enter drying machine 262 (such as freeze drier) afterwards, pulverizer 264 obtains high-purity alpha -ol after crushing Molten protein product.Enter in filter 256 through material in filter 266, retention material enters drying after washing (268) Device 270 (as restrain dry and) dry, and by pulverizer 272 (such as beater grinder) crushing after protein feed product.Filtering The transmission feed liquid of machine 258 and 266 is discharged into water treatment system.

According to exemplary embodiment, filter 258 can have the pore size filter of such as 500 mesh.

The running parameter of zeins extraction is listed in table 1 below, it provides typical model for each operating procedure Enclose, preferred scope.A typical range of from 2~150 μm of raw material powder particle diameter.The preferred scope of raw material powder particle diameter is 10~100 μ M (particle diameter with mesh number can table look-up conversion).A typical range of from aqueous the 50%~95% of the moisture regulation of raw material.The moisture of raw material is adjusted The preferred scope of section is aqueous 75%~90%.The 0.01wt% of protein by weight in a typical range of from raw material of protease addition ~10wt%.The preferred scope of protease addition is 0.1wt%~3wt% of protein by weight in raw material.Protease hydrolyzed pH A typical range of from 3.5~10.5.Protease hydrolyzed pH preferred scope is 3.8~10.The typical model of protease hydrolyzed temperature Enclose for 20 DEG C~65 DEG C.The preferred scope of protease hydrolyzed temperature is 35 DEG C~55 DEG C.The typical range of protease hydrolyzed time For 0.2h~10h.The preferred scope of protease hydrolyzed time is 0.5h~5h.In a typical range of from raw material of amylase addition 0.05wt%~20wt% of starch weight.The preferred scope of amylase addition be raw material in starch weight 0.25wt%~ 15wt%.A typical range of from the 3~8 of amylase enzymolysis pH.Amylase enzymolysis pH preferred scope is 3.3~7.5.Amylase enzyme Solve a typical range of from 30 DEG C~72 DEG C of temperature.The preferred scope of amylase enzymolysis temperature is 35 DEG C~63 DEG C.Amylase enzymolysis A typical range of from 1h~10h of time.The preferred scope of amylase enzymolysis time is 2h~7h.The allusion quotation of fiber hydrolase addition Type scope is 0.2wt%~30wt% of fibre weight in raw material.The preferred scope of fiber hydrolase addition is fibre in raw material Tie up 0.5wt%~20wt% of weight.A typical range of from the 4~6.5 of fiber hydrolase pH.Fiber hydrolase pH preferred scope For 4.5~5.5.A typical range of from 40 DEG C~60 DEG C of fiber hydrolase temperature.The preferred scope of fiber hydrolase temperature is 45 DEG C~55 DEG C.A typical range of from 0.5h~12h of fiber hydrolase time.The preferred scope of fiber hydrolase time be 2h~ 8h.Alkali includes sodium hydroxide, potassium hydroxide.Alkali addition is determined according to reaction pH.Acid includes hydrochloric acid, sulfuric acid, sulfurous acid, organic Sour (including lactic acid, citric acid, malic acid).Sour addition is determined according to reaction pH.Reagent composition in system is in feed liquid The addition situation molar concentration of water section contained by feed liquid (concentration be)：A typical range of from 1mM of sulfur-containing compound addition~ 50mM.The preferred scope of sulfur-containing compound addition is 5mM~30mM.A typical range of from 0.5mM of phosphorus-containing compound addition ~60mM.The preferred scope of phosphorus-containing compound addition is 2mM~40mM.Sodium ion, potassium ion, divalent alkaline-earth metal ion, A typical range of from 0.01mM~20mM of the addition of divalent transition metal ion.Sodium ion, potassium ion, divalent alkaline-earth metal from Son, the preferred scope of the addition of divalent transition metal ion are 0.1mM~12mM.The typical range of metal-chelator addition For 0mM~35mM.The preferred scope of metal-chelator addition is 0.1mM~18mM.Operating pressure typical case model in reaction vessel Enclose for -0.1MPa~0.3MPa.A typical range of from 200g~150000g of centrifugal force.The preferred scope of centrifugal force be 1000g~ 7000g.A typical range of from 1 μm~80 μm of pore size filter.The preferred scope of pore size filter is 10 μm~50 μm.Membrane filtration aperture A typical range of from 10nm~10um.The preferred scope in membrane filtration aperture is 20nm~1 μm.

In the hydrolyzation system of the present invention, acid, alkali make reaction be in suitable soda acid ring as pH adjusting agent for adjusting Border, and it is also adjusted for the dissolved state of component.Sulfur-containing compound, phosphorus-containing compound and metal ion are mainly used in adjusting egg The structure of white background thing, specifically it act as opening the disulfide bond in protein substrate, therefore can be closed in theory from any The suitable compound that can open the disulfide bond in albumen.Metal ion and chelating agent are used for regulatory protein enzyme, carbohydrate The activity or stability of the enzymes such as enzyme.

Table 1：Running parameter table for extracting alcohol soluble protein

It should be pointed out that described in following examples extraction raw material composition and albumen finished product composition etc. numerical value simultaneously It is not used in and involved extraction raw material and albumen finished product is defined to specific specific products, and is only for description and extracts raw material Or the objective parameter of albumen finished product.

In the present invention, according to《The measure of national food safety standard GB5009.5-2010-- Protein in Food》In " the first method " determine product in protein content, protein conversion factor is 6.24.Determined and produced according to GB/T5009.6 method Fat content in product.Moisture in product is determined according to GB/T5009.3 method.According to GB5009.12, GB5009.17 Lead content in product is determined with GB5009.11 method.According to method determine product in mercury content.According to method determine production Arsenic content in product.Aflatoxin content in product is determined according to GB5009.23 method.According to GB4789.2, GB4789.3, GB4789.15 and GB4789.4 method determines total plate count in product, coliform, mould and the amount of pathogenic bacteria respectively.

In the present invention, the OD value (as shown in Figure 4) of protein is determined by the following method：Concentration is used for 15% (w/v) separation gel carries out reproducibility sds polyacrylamide to the prepared sample added with 10% (v/v) beta -mercaptoethanol Gel electrophoresis, coomassie brilliant blue staining (bottom graph in Fig. 4 A-E) is carried out to the gel after electrophoresis.Use Quantity One Software carries out photodensitometry to band in gel can produce signal peak (upper diagram in Fig. 4 A-E).The area of signal peak is should The OD value of component protein is in functional relation with protein content.Specifically, by the OD value of different Gliadin Components Be designated as Di (i=1,2,3 ..., n), the ratio between percentage OD value of alcohol soluble protein are accounted for alcohol soluble protein each component and is calculated, That is Di/D0%.

According to exemplary embodiment, the extraction raw material containing alcohol soluble protein used in the present invention can have such as Fig. 4 A Electrophoretic image and its optical density collection of illustrative plates comprising alpha-alcohol soluble protein, the molten albumen of β -ol, the molten albumen of γ -ol and the molten albumen of δ -ol.

According to exemplary embodiment, alcohol soluble protein product of the invention can have such as Fig. 4 C's to include the molten egg of α -ol White electrophoretic image and its optical density collection of illustrative plates.

According to exemplary embodiment, alcohol soluble protein product of the invention can have such as Fig. 4 D's to include the molten egg of α -ol In vain, the electrophoretic image and its optical density collection of illustrative plates of the molten albumen of β -ol and the molten albumen of γ -ol.

According to exemplary embodiment, alcohol soluble protein product of the invention can have such as Fig. 4 A's to include the molten egg of α -ol The electrophoretic image and its optical density collection of illustrative plates of the white and molten albumen of β -ol.

According to exemplary embodiment, alkali protease used in the present invention is the serine stretch protein of bacillus Enzyme.According to exemplary embodiment, acid protease used in the present invention is mould carboxyl protease.According to exemplary Embodiment, neutral proteinase used in the present invention is the metalloproteinases of mould or bacillus.According to exemplary reality Mode is applied, thiol protease used in the present invention is the mercapto from plant (stem end, leaf, skin and the pawpaw fruit of such as pineapple) Base protease, such as bromelain, papain.

In upper table 1 in listed reagent composition, acid, alkali, which are used for regulation, makes reaction be in suitable acid or alkali environment, and And it is also adjusted for the dissolved state of component.Sulfur-containing compound, phosphorus-containing compound and metal ion are mainly used in regulatory protein bottom The structure of thing；Metal ion and chelating agent are used for the activity or stability of the enzymes such as regulatory protein enzyme, carbohydrase.This area Technical staff had the ability according to the teachings of the present invention, completely understanding should how according to used in actual enzyme and corresponding bottom Thing selects suitable reagent composition and its concentration, to realize the purpose of the present invention (for example, those skilled in the art can fit Locality is selected enzyme processing time, and passes through ordinary skill Knowl- edge Control hydrolysis time so that substrate reaches institute Desired hydrolysis degree, to realize that the utilization particle diameter difference filtering that the present invention is emphasized removes hydrolysate).

Embodiment

Unless defined otherwise, the percentage composition indicated in the present invention is weight percentage.In following examples In, if only indicating concentration when adding related reagent, then it represents that the reagent rear concentration reached in system is added to.Such as nothing Special instruction, all reactions are carried out at ambient pressure.

Embodiment 1

Adjusted enzymatic vessel is squeezed into after the maize yellow-powder regulation moisture of aqueous 8.9%, albumen 64% (butt) to 70% Section to pH4.8,45 DEG C, and add and account for 4.8% acid protease of contained protein content (MA-SD, A Manuo amano enzyme preparation have Limit company), 60mM tri- (2- carboxyethyls) phosphine reaction adjusted after 1.2 hours to pH7.5,52 DEG C, addition 2.6% alkali protease (SUKAPro NE, Su Kehan bioengineering shares have for (2709, Pang Bo bioengineering Co., Ltd) and 2% neutral proteinase Limit company), 1mM mercaptoethanols react 0.5 hour after centrifuge, wash and collect precipitation.Total protein contents on dry basis in products obtained therefrom 61.1%, the content of prolamine in albumen is more than 74%, and alpha-alcohol soluble protein content is that the molten protein content of 95%, β -ol is 2%.

Embodiment 2

By the maize yellow-powder of aqueous 62.4%, protein 70 % (butt) add water regulation to squeeze into after 75% enzymatic vessel adjust to PH3.8,35 DEG C, and add acid protease (the SUKAPro AC Su Kehan bioengineering shares of contained protein by weight 0.38% Co., Ltd), 0.26% bromelain (food-grade, Pang Bo bioengineering Co., Ltd), 50mM sodium pyrosulfites, 0.5mM tri- (2- carboxyethyls) phosphine, 4mM manganese ions first react 10 hours, then be warming up to 50 DEG C maintain 0.5 hour, then adjust to PH8.3,52 DEG C, and add 0.33% alkali protease (2709, Pang Bo bioengineering Co., Ltd) react 1 hour after add 10mM EDTA are simultaneously centrifuged, and are washed and are collected precipitation.The molten egg of alcohol in products obtained therefrom in total protein contents on dry basis 54.1%, albumen Bai Hanliang 97.55%, wherein containing 95.85% alpha-alcohol soluble protein, the 1.9% molten albumen of β -ol and the 0.95% molten egg of γ -ol In vain.

Embodiment 3

After the maize yellow-powder material regulation moisture of aqueous 89.9%, albumen 68% (butt) to 80%, enzyme is squeezed into Solution tank adjust to pH6.5,45 DEG C, (food-grade, Pang Bo bioengineering have the papain of the contained protein by weight 0.01% of addition Limit company), after 20mM mercaptoethanols and 0.01mM cobalt ions reacts 1.5 hours, then adjust to pH10.2,45 DEG C, and add 0.01% alkali protease (Protex 6L, Jie Nengke bioengineering Co., Ltd) is centrifuged after reacting 2 hours, is washed and is received Collection precipitation.Content of prolamine in products obtained therefrom in total protein contents on dry basis 55.9%, albumen is more than 90.66%, wherein containing 85.54% alpha-alcohol soluble protein, the 9.91% molten albumen of β -ol and the 4.34% molten albumen of γ -ol.

Embodiment 4

Enzymatic vessel will be squeezed into after aqueous 9.0%, protein 28 % (butt) distiller's dried grain (DDG) regulation moisture to 80%, PH4.2, acid protease (the limited public affairs of MA-SD, A Manuo amano enzyme preparation that contained protein by weight 0.8% is added at 35 DEG C Department), 40mM tri- (2- carboxyethyls) phosphine reaction 6 hours, regulation pH to 6.5,45 DEG C, add 0.5% neutral proteinase (1398, Pang Bo bioengineering Co., Ltd) and 1.7% papain (food-grade, Pang Bo bioengineering Co., Ltd), 5mM sulfydryls Ethanol, 0.1mM zinc ions are centrifuged after reacting 0.5 hour, are washed and are collected precipitation.Total protein contents on dry basis in products obtained therefrom 23.9%, the content of prolamine 81.30% in albumen, wherein the molten egg of β -ol containing 94.5% alpha-alcohol soluble protein, 1.95% The white molten albumen of γ -ol with 0.04%.

Embodiment 5

The aqueous 11.0%, distiller's dried grain of albumen 30% (butt) and DDGS (DDGS) are added water after modulating and washing twice Moisture is adjusted to 95%, contained protein by weight 0.23wt% acid protease (SUKAPro is added at pH4.8,55 DEG C AC Su Kehan bioengineering limited company) and 2mM tri- (2- carboxyethyls) phosphine reaction after 0.5 hour, at pH4.8,53 DEG C The bromelain (food-grade, Pang Bo bioengineering Co., Ltd) of the contained protein by weight 0.3% of addition reacts 2.8 hours, then Adjust pH to 7.2 and at 53 DEG C add 0.5% neutral proteinase (1398, Pang Bo bioengineering Co., Ltd), 6mM calcium from Son, 15mM mercaptoethanols and 15mM cysteines are centrifuged after reacting 3.1 hours, are washed and are collected precipitation.Total egg in products obtained therefrom It is molten containing 87.5% alpha-alcohol soluble protein, the 3.1% molten albumen of β -ol and 5.1% γ -ol in white contents on dry basis 24.3%, albumen Albumen.

Embodiment 6

Aqueous 31.0%, protein 32 % (butt) wet vinasse (WDG) are added water into regulation moisture to 80%, PH7.5, neutral proteinase (SUKAPro NE, Su Kehan the bioengineering stock that contained protein by weight 0.04wt% is added at 25 DEG C Part Co., Ltd), 2mM magnesium ions, 2mM calcium ions and 20mM mercaptoethanols react 4.5 hours, then adjust the maintenances of pH to 10.1 0.2 hour.The first permeate and trapped fluid are obtained using 1 μm of aperture membrane filtration, regulation trapped fluid moisture returns enzyme to 80% Solution tank regulation pH to 10.1, (2709, Pang Bo is biological for the alkali protease for the contained protein by weight 0.02wt% that adds materials after 55 DEG C Engineering Co., Ltd) insulation 0.5 hour after centrifuge, wash and collect precipitation.Total protein contents on dry basis 25.7% in products obtained therefrom, In albumen content of prolamine be more than 93.1%, wherein containing 77.9% alpha-alcohol soluble protein, the 17.2% molten albumen of β -ol and The 1.3% molten albumen of γ -ol.

Embodiment 7-12 is using maize yellow-powder of the water content for 80% and the contained albumen weight of addition at pH4.2,35 DEG C Acid protease (MA-SD, A Manuo amano enzyme preparation Co., Ltd), 10mM tri- (2- carboxyethyls) phosphine reaction 3 of amount 2% are small When, regulation pH to 6.5,45 DEG C, add 0.5% neutral proteinase (1398, Pang Bo bioengineering Co., Ltd) and 1.7% Papain (food-grade, Pang Bo bioengineering Co., Ltd) react 1 hour after material.

Embodiment 7

Above-mentioned material is crushed after (about 40 μm of abrasive disk space) with colloid mill (MagicLab, IKA company), enzymatic vessel is squeezed into Regulation to pH5.0,63 DEG C, and add starch contained therein weight 8wt% alpha amylase (Spezyme Fred, Jie Neng sections bioengineering Co., Ltd) and 4% carbohydrase (Spirizyme Ultra, letter enzyme preparation Co., Ltd of Novi) reaction 2 hours；200g from Heart 15min (ALLEGRA 30R, Beckman Coulter Inc. of the U.S.), collect precipitation regulation moisture to 70% then Squeeze into the second enzymatic vessel adjust to pH5.5,50 DEG C, and add 3.0% cellulase (SUKAZYM- of contained fibre weight SUKACell, Su Kehan bioengineering limited company) and 2% include arabanase, cellulase, beta glucan The complex enzymes (Viscozyme L, letter enzyme preparation Co., Ltd of Novi) of the enzymes such as enzyme, hemicellulase, zytase, 10mM calcium from Son, 10mM potassium ions react 2 hours after, using 50 μm of aperture filter-cloth filterings (protein-contg about 100 μm of target phase average particle diameter, Both less than 30 μm greatly of the average grain diameter of non-targeted phase particle diameter different component (containing DDGS)) obtain the first filter cake, adjusted to Moisture 70%, pH7.5,55 DEG C, and add after 35mM EDTA, pass through 10 μm of membrane filtrations (protein-contg target phase average grain About 15 μm of footpath, non-targeted mutually mostly be DDGS) after obtain the second filtrate and filter cake, to gained filter cake by washing, dry after Obtain the zein product that dry protein content is 88.4%.

Embodiment 8

Above-mentioned material is crushed into the regulation that adds water after (about 2 μm of abrasive disk space) with colloid mill (MagicLab, IKA company) to arrive 75% squeeze into the first enzymatic vessel adjust to pH3.0,35 DEG C, and add starch contained therein weight 1wt% alpha amylase (Liquozyme Letter enzyme preparation Co., Ltd of SCDS Novi) and composite alpha-amylase (Novozyme NSs of the 1wt% comprising carbohydrase and Pullulanase 50013, letter enzyme preparation Co., Ltd of Novi) react 7 hours, use 1 μm of aperture strainer filtering (protein-contg target phase average grain About 150 μm of footpath, both less than 2 μm greatly of the average grain diameter (containing DDGS) of non-targeted phase different component) the first filter cake is obtained, adjust water Divide content to 75%；Material squeeze into the 3rd enzymatic vessel adjust to pH6.5,45 DEG C, and add fibre weight 12% cellulase, 5% 1,4 beta-glucanase (letter enzyme preparation Co., Ltd of Ultraflo Novi, mainly comprising 1,4 beta-glucanase, also comprising xylan Enzyme etc.) and 13% complex enzyme (Viscozyme L Novi letter enzyme preparation Co., Ltd) reaction 4 hours after addition 10mM EDTA And pH to 5.0 is adjusted, using 40 μm of aperture filter-cloth filterings, (protein-contg about 110 μm of target phase average particle diameter, non-targeted is mutually different Average big both less than 30 μm (the containing DDGS) of component) the second filtrate and filter cake are obtained, filter cake obtains egg after washing, drying The zein product that white contents on dry basis is 99.2%.

Embodiment 9

Above-mentioned material is crushed after (about 10 μm of abrasive disk space) with colloid mill (MagicLab, IKA company), the first enzyme is squeezed into Solution tank is adjusted to pH4.0,40 DEG C, the contained fibre weight 0.2wt% of addition cellulase (Celluclast, Novi's letter enzyme system Agent Co., Ltd), 0.4% complex enzyme (letter enzyme preparation Co., Ltd of Viscozyme L Novi) and 0.01mM potassium ion it is anti- Answer 8 hours.Using 80 μm of aperture filter-cloth filterings (protein-contg about 140 μm of target phase average particle diameter, non-targeted phase different component Both less than 9 μm greatly of average grain diameter (containing DDGS)) the first filtrate and filter cake are obtained, the first filter cake adjusts moisture to 80% Afterwards, squeeze into the second enzymatic vessel adjust to pH6.5,45 DEG C, add the alpha amylase (Spezyme of starch contained therein weight 0.1% Fred, Jie Nengke bioengineering Co., Ltd), (Spirizyme Ultra, Novi's letter enzyme preparation is limited for 0.05% carbohydrase Company), 0.1% Pullulanase (letter enzyme preparation Co., Ltd of Promozyme D2 Novi), and be warming up to 72 DEG C and maintain 5 hours After adjust to pH10.5 using 20nm apertures membrane filtration that (protein-contg about 0.5 μm of target phase average particle diameter, non-targeted is mutually big All be DDGS) obtain the second permeate and trapped fluid, trapped fluid is washed, dehydrate after obtain dry protein content and be 85.7% zein product.

Embodiment 10

By above-mentioned material with colloid mill (MagicLab, IKA company) crush (about 100 μm of abrasive disk space) afterwards adjust moisture to 80%, the first enzymatic vessel is squeezed into, contained fibre weight 0.2wt% complex cellulase is added at pH4.5,60 DEG C (letter enzyme preparation Co., Ltd of Viscozyme L Novi), 0.1mM EDTA, reaction is centrifuged after 12 hours, after washing once Moisture is adjusted to 80%；Adjust the composite alpha-amylase (Spirizyme that pH to 8 adds starch contained therein weight 0.05% at 45 DEG C Excel, letter enzyme preparation Co., Ltd of Novi) using 80 μm of aperture filter-cloth filterings, (protein-contg target is equal after 10 hours for maintenance Equal about 5 μm of particle diameter, both greater than 90 μm greatly of the average grain diameter of non-targeted phase different component) obtain the first filtrate and filter cake, the first filter Liquid obtains the by 100nm microfiltration membranes (protein-contg about 5 μm of target phase average particle diameter, non-targeted is mostly mutually DDGS) again Two filtrates.Second trapped fluid, which centrifuges 10min under 1000g and obtains precipitating, to be obtained albumen butt after washing, dehydrating and contains Measure the zein product for 75.8%.

Embodiment 11

Above-mentioned material is crushed after (about 150 μm of abrasive disk space) with colloid mill (MagicLab, IKA company), first is squeezed into Enzymatic vessel, (Celluclast, Novi's letter enzyme preparation has the contained fibre weight 12wt% of addition cellulase at pH5,55 DEG C Limit company), 8wt% 1,4 beta-glucanase (letter enzyme preparation Co., Ltd of Ultraflo Novi), 12mM calcium ions, reaction 0.5 is small When；The is obtained using 10 μm of aperture membrane filtrations (protein-contg about 110 μm of target phase average particle diameter, non-targeted is mutually DDGS) One permeate and trapped fluid, regulation trapped fluid moisture is to squeezing into the second enzymatic vessel after 85%, then adjusts pH to 7.5 and at 50 DEG C Add alpha amylase (Spezyme Fred, Jie Nengke bioengineering Co., Ltd), 1% carbohydrase of starch contained therein weight 1% (Spirizyme Ultra, letter enzyme preparation Co., Ltd of Novi) and 3% composite alpha-amylase (Spirizyme Excel, Novi Believe enzyme preparation Co., Ltd) using 50 μm of aperture filter-cloth filterings, (protein-contg target is equal after 3.5 hours for 18mM EGTA reactions About 15 μm of equal particle diameter and DDGS, both greater than 140 μm greatly of the average grain diameter of non-targeted phase different component) obtain the second permeate and Filter cake, permeate, which centrifuges solid phase obtained by 10min under 2000g and obtains dry protein content after washing, dehydrating, is 80.0.% zein product.

Embodiment 12

By above-mentioned crushing material particle diameter to about 30 μm, the first enzymatic vessel is squeezed into, starch contained therein is added at pH3.5,30 DEG C Weight 5wt% alpha amylase (Liquozyme SCDS, letter enzyme preparation Co., Ltd of Novi), 3wt% carbohydrase (Spirizyme Ultra, letter enzyme preparation Co., Ltd of Novi), (Promozyme D2 Novi believes enzyme system to 2wt% Pullulanase Agent Co., Ltd), 10% composite alpha-amylase (Novozyme NS50013, letter enzyme preparation Co., Ltd of Novi), 2mM calcium ions it is anti- Answer 1 hour；Then regulation pH to 5.6 and added at 50 DEG C contained fibre weight 10% composite fibre hydrolase (GC 518, Jie Nengke bioengineering Co., Ltd) react 5 hours；Using 1 μm of aperture membrane filtration, (protein-contg target phase average particle diameter is big Both greater than 100 μm, non-targeted is mutually DDGS) the first permeate and trapped fluid are obtained, regulation trapped fluid moisture is squeezed into 80% Size mixing tank, regulation pH to 10, by 20 μm of filter screen, (protein-contg about 1 μm of target phase average particle diameter, non-targeted is mutually not after 55 DEG C It is about 29 μm and DDGS with the average grain diameter of component) the second filtrate and trapped fluid are obtained, obtain egg after filtrate is dehydrated The zein product that white contents on dry basis is 95.0%.

Raw material in embodiment 13,14 uses aqueous 80% maize yellow-powder, adjusts pH to 5.6 and is added at 50 DEG C The composite fibre hydrolase of contained fibre weight 8% and 2.0% cellulase react 2 hours, or make drop with other conditions The fibre weight of solution accounts for 1/10th of fibrillation amount.

Embodiment 13

Above-mentioned material is 20 μm of colloid mill by abrasive disk space, and the pH of grinding material is adjusted to 3,4 and respectively 6.2, and obtain filtering trapped fluid using aperture for 10 μm of filter membrane, and after trapped fluid is centrifuged and dried, respectively obtain albumen Content is 71.25%, 70.76% and 70.36% product；When obtaining filter cake using aperture for 48 μm of strainer filtering, and will After cake dewatering is dried, the product that protein content is 77.7%, 76.8% and 76.3% is respectively obtained.It is 0.1 μ when using aperture M filter membrane or aperture are 75 μm, 150 μm of filter screen is handled after the material of pH4 in the present embodiment, and gained trapped fluid or filter cake enter One step is dehydrated and dried, and respectively obtains the protein product that protein content is 69.9%, 76.4% and 71.9%.

Embodiment 14

The pH of above-mentioned material is adjusted to 6.9,8 and 10.5 respectively, and permeate is obtained for 1 μm of filter membrane using aperture, It will transmit through after liquid centrifuges and dry, respectively obtain the protein product that protein content is 93.2%, 96.0% and 97.5%；When using Aperture obtains filter cake for 75 μm of strainer filtering, and after cake dewatering is dried, respectively obtain protein content for 76.9%, 78.8% and 79.7% product.When handling this reality using the filter screen that the filter membrane or aperture that aperture is 0.1 μm are 38 μm, 150 μm After the material for applying pH8 in example, gained trapped fluid or filter cake are further dehydrated and dried, and respectively obtaining protein content is 99.5%th, 87.2% and 72.2% protein product.

Raw material in embodiment 15,16 uses aqueous 80% maize yellow-powder, regulation to pH6.5,45 DEG C, adds institute Alpha amylase, 1% carbohydrase, 2% Pullulanase of starch-containing weight 2%, and be warming up to 72 DEG C and maintain 2.5 hours, or make Liquefied amount of starch is accounted for the half of ative starch with other conditions, adjust pH to 5.6 and institute's fibre-bearing weight is added at 50 DEG C The composite fibre hydrolase of amount 8% and 2.0% cellulase react 2 hours, or make with other conditions the fibre weight of degraded Account for 1/10th of fibrillation amount.

Embodiment 15

Above-mentioned material is 20 μm of colloid mill by abrasive disk space, and the pH of grinding material is adjusted to 3,4 and respectively 6.2, and obtain filtering trapped fluid using aperture for 10 μm of filter membrane, and after trapped fluid is centrifuged and dried, respectively obtain albumen Content is 79.0%, 78.7% and 78.3% product；When obtaining filter cake using aperture for 48 μm of strainer filtering, and by filter cake After dehydrating, the product that protein content is 83.6%, 83.0% and 82.63% is respectively obtained.It it is 0.1 μm when using aperture Filter membrane or aperture are 75 μm, 150 μm of filter screen is handled after the material of pH4 in the present embodiment, and gained trapped fluid or filter cake enter one Step is dehydrated and dried, and respectively obtains the protein product that protein content is 78.0%, 82.7% and 79.4%.

Embodiment 16

The pH of above-mentioned material is adjusted to 6.9,8 and 10.5 respectively, and permeate is obtained for 1 μm of filter membrane using aperture, It will transmit through after liquid centrifuges and dry, respectively obtain the protein product that protein content is 95.9%, 97.4% and 98.2%；When using Aperture obtains filter cake for 75 μm of strainer filtering, and after cake dewatering is dried, respectively obtain protein content for 84.8%, 86.1% and 86.8% product.When handling this reality using the filter screen that the filter membrane or aperture that aperture is 0.1 μm are 38 μm, 150 μm After the material for applying pH8 in example, gained trapped fluid or filter cake are further dehydrated and dried, and respectively obtaining protein content is 99.5%th, 92.3% and 80.0% protein product.

Embodiment 17

By aqueous 8.9% maize yellow-powder, using airslide disintegrating mill, (FQS15 types, Shanghai causes triumphant powder machinery manufacture limited Company) powder particle diameter is to the regulation that adds water after about 40 μm to after 50% abundant aquation, into enzymolysis and piece-rate system：Material enters first Enter the first enzymatic vessel adjust to pH5.0,63 DEG C, and add starch contained therein weight 8wt% alpha amylase (Spezyme Fred, it is outstanding Neng Ke bioengineering Co., Ltd) and 4% carbohydrase (Spirizyme Ultra, letter enzyme preparation Co., Ltd of Novi) reaction 2 Hour；200g centrifugation 15min (ALLEGRA 30R, Beckman Coulter Inc. of the U.S.), collect precipitation regulation moisture To 70% then squeeze into the second enzymatic vessel adjust to pH5.5,50 DEG C, and add 3.0% cellulase of contained fibre weight (SUKAZYM-SUKACell, Su Kehan bioengineering limited company) and 2% complex enzyme (Viscozyme L, Novi's letter Enzyme preparation Co., Ltd), 10mM calcium ions, 10mM potassium ions react 2 hours after, obtain first using 50 μm of aperture filter-cloth filterings Filter cake, adjusted moisture to squeeze into after 70% the 3rd enzymatic vessel adjust to pH4.8,35 DEG C, and add contained protein content 5% acid protease (carboxyl protease MA-SD, A Manuo amano enzyme preparation Co., Ltd), 60mM tri- (2- carboxyethyls) Phosphine reaction is adjusted after 1.5 hours to pH7.5,55 DEG C, alkali protease (serine protease 2709, Pang Bo biologies of addition 3% Engineering Co., Ltd) and 2% neutral proteinase (the limited public affairs of metalloproteinases SUKAPro NE, Su Kehan bioengineering shares Department), 1mM mercaptoethanols react 0.5 hour after add 35mM EDTA, by obtaining the second filtrate and filter cake after 10 μm of membrane filtrations. Gained filter cake washed, dry after the alcohol that obtains in the zeins product that dry protein content is 90%, albumen it is molten Protein content is more than 74%, and all alpha-alcohol soluble proteins (100%), and fat and ash content contents on dry basis are respectively 1.04% He 4.01%.In addition, micro-filtrate membrane filtration of second filtrate by 1 μm, trapped fluid, which refines and albumen butt is obtained after dehydrating, to be contained The product of amount 86%.

Embodiment 18

Aqueous 62.4% maize yellow-powder is used into roll squeezer (S120, Changzhou rely on oneself chemical machinery Co., Ltd) crushing Add water and adjusted to after 75% after (about 2 μm of roller gap), into enzymolysis and piece-rate system：Material initially enters the first enzymatic vessel tune Section to pH3.0,35 DEG C, and add starch contained therein weight 1wt% alpha amylase (Liquozyme SCDS Novi letter enzyme preparation have Limit company) and composite alpha-amylase (Novozyme NS 50013, Novi letter enzyme preparation of the 1wt% comprising carbohydrase and Pullulanase Co., Ltd) react 7 hours, the first filter cake is obtained using 1 μm of aperture strainer filtering, moisture is adjusted to 75%；Beat Enter the second enzymatic vessel adjust to pH3.8,35 DEG C, and add the acid protease (carboxyl protease of contained protein by weight 0.4% SUKAPro AC Su Kehan bioengineering limited company), (food-grade, Pang Bo bioengineering have 0.3% bromelain Limit company), 50mM sodium pyrosulfites, 0.5mM tri- (2- carboxyethyls) phosphine, 4mM manganese ions first react 10 hours, then be warming up to 50 DEG C Maintain 0.5 hour, then adjust to pH8.5,65 DEG C, and add 0.3% alkali protease (serine protease 2709, it is huge Rich bioengineering Co., Ltd) 150000g centrifugation 10s (Optima after reaction 1 hour^TMXE, the limited public affairs of U.S.'s Beckman Kurt Department), precipitation regulation moisture is collected to 75%；Material squeeze into the 3rd enzymatic vessel adjust to pH6.5,45 DEG C, and add fiber The cellulase of weight 12%, 5% 1,4 beta-glucanase (Ultraflo Novi letter enzyme preparation Co., Ltd) and 13% are combined Enzyme (letter enzyme preparation Co., Ltd of Viscozyme L Novi) reaction is added 10mM EDTA and adjusted to pH to 5.0, makes after 4 hours The second filtrate and filter cake are obtained with 40 μm of aperture filter-cloth filterings, filter cake obtains dry protein content after washing, drying and is Content of prolamine in 99.1% zeins product, albumen is more than 98.4%, wherein containing the 97% molten egg of α -ol In vain, the 2% molten albumen of β -ol and the 1% molten albumen of γ -ol, fat and ash content contents on dry basis are respectively 0.5% and 0.2%.Separately Outside, the second filtrate is by 1 μm of micro-filtrate membrane filtration, and trapped fluid refines and dry protein content 75.1% is obtained after dehydrating Product.

Embodiment 19

Aqueous 89.9% maize yellow-powder is crushed into (the μ of abrasive disk space about 10 using colloid mill (MagicLab, IKA company) M), enzymolysis and piece-rate system are squeezed into：Material initially enters the first enzymatic vessel and adjusted to pH4.0,40 DEG C, adds institute's fibre-bearing weight Measure 0.2wt% cellulase (Celluclast, letter enzyme preparation Co., Ltd of Novi), 0.4% complex enzyme (Viscozyme Letter enzyme preparation Co., Ltd of L Novi) and 0.01mM potassium ion reaction 8 hours.First is obtained using 80 μm of aperture filter-cloth filterings Filtrate and filter cake, the first filtrate obtain the second filtrate by 1 μm of microfiltration membranes again, and the second filtrate is obtained by 10nm milipore filter To the 3rd trapped fluid, the first filter cake and the 3rd trapped fluid are merged and moisture is adjusted to after 80%, the second enzymatic vessel is squeezed into Regulation to pH8.0,45 DEG C, (Protex 6L, Jie Neng sections bioengineering has the alkali protease of the contained protein by weight 0.4% of addition Limit company), 20mM mercaptoethanols react 1.5 hours, add the alpha amylase (Spezyme of starch contained therein weight 0.1% Fred, Jie Nengke bioengineering Co., Ltd), (Spirizyme Ultra, Novi's letter enzyme preparation is limited for 0.05% carbohydrase Company), 0.1% Pullulanase (letter enzyme preparation Co., Ltd of Promozyme D2 Novi), and be warming up to 72 DEG C and remain 5 small When；Material is squeezed into the 3rd enzymatic vessel and adjusted to pH10.5,45 DEG C of reactions 0.5 hour obtains the using 20nm apertures membrane filtration Four permeate and trapped fluid, trapped fluid is washed, dehydrate after obtain dry protein content be 85% zeins Content of prolamine in product, albumen is more than 92%, wherein the molten albumen of β -ol and 2% containing 91% alpha-alcohol soluble protein, 7% The molten albumen of γ -ol, fat and ash content contents on dry basis be respectively 3.2% and 1.52%.In addition, being obtained after the 4th permeate is refined The product of dry protein content 94.7%.

Embodiment 20

By aqueous 9.0% distiller's dried grain (DDG), using airslide disintegrating mill, (FQS15 types, Shanghai causes triumphant powder machinery manufacture to have Limit company) powder particle diameter to regulation moisture after about 100 μm, to 80%, squeezes into enzymolysis and piece-rate system：Material initially enters first Enzymatic vessel, adds contained fibre weight 0.2wt% complex cellulase (Viscozyme L Novi letter at pH4.5,60 DEG C Enzyme preparation Co., Ltd), 0.1mM EDTA, reaction centrifuges after 12 hours, washing once afterwards regulation moisture to 80%；By thing Material squeezes into the second enzymatic vessel, and acid protease (MA-SD, the A Manuo days of contained protein by weight 1% are added at pH4.2,35 DEG C Wild enzyme preparation Co., Ltd), 40mM tri- (2- carboxyethyls) phosphine reaction 6 hours, regulation pH to 6.5,45 DEG C added in 0.5% (food-grade, Pang Bo bioengineering have for property protease (1398, Pang Bo bioengineering Co., Ltd) and 1.5% papain Limit company), 5mM mercaptoethanols, 0.1mM zinc ions react 0.5 hour after regulation pH to 8 45 DEG C add starch contained therein weight 0.05% composite alpha-amylase (Spirizyme Excel, letter enzyme preparation Co., Ltd of Novi) uses 80 μm after maintaining 10 hours Aperture filter-cloth filtering obtains the first filtrate and filter cake, and the first filtrate obtains the second filtrate by 100nm microfiltration membranes again.Second section Stay liquid to centrifuge 10min under 1000g to obtain precipitating the corn for obtaining that dry protein content is 75% after washing, dehydrating Content of prolamine in alcohol soluble protein product, albumen is more than 81.50%, wherein containing 98% alpha-alcohol soluble protein, 1.99% The molten albumen of β -ol and the 0.03% molten albumen of γ -ol, fat and ash content contents on dry basis are respectively 4.98% and 2.11%.In addition, Second permeate and the first filter cake merge and the product of dry protein content 50% are obtained after drying.

Embodiment 21

By aqueous 11.0% distiller's dried grain and DDGS (DDGS) add water modulate and wash twice afterwards regulation moisture to 95%, (about 150 μm of abrasive disk space) is crushed using colloid mill (MagicLab, IKA company), enzymolysis and piece-rate system is squeezed into：Thing Material initially enters the first enzymatic vessel, added at pH5,55 DEG C contained fibre weight 12wt% cellulase (Celluclast, Letter enzyme preparation Co., Ltd of Novi), 8wt% 1,4 beta-glucanase (letter enzyme preparation Co., Ltd of Ultraflo Novi), 12mM calcium Ion, contained protein by weight 0.2wt% acid protease (SUKAPro AC Su Kehan bioengineering limited company) and 2mM tri- (2- carboxyethyls) phosphine) react 0.5 hour；The first permeate and trapped fluid are obtained using 10 μm of aperture membrane filtrations, is adjusted Trapped fluid moisture adds the bromelain of contained protein by weight 0.3% to the second enzymatic vessel is squeezed into after 85% at pH5,50 DEG C Enzyme (food-grade, Pang Bo bioengineering Co., Ltd) reacts 3 hours, then adjusts pH to 7.5 and added at 50 DEG C in 0.5% Property protease (1398, Pang Bo bioengineering Co., Ltd), starch contained therein weight 1% alpha amylase (Spezyme Fred, it is outstanding Neng Ke bioengineering Co., Ltd), 1% carbohydrase (Spirizyme Ultra, letter enzyme preparation Co., Ltd of Novi) and 3% Composite alpha-amylase (Spirizyme Excel, letter enzyme preparation Co., Ltd of Novi), 15mM mercaptoethanols and 15mM cysteines The second permeate and filter cake are obtained using 50 μm of aperture filter-cloth filterings after being reacted 3.5 hours with 18mM EGTA, permeate exists 2000g descends to centrifuge 10min gained solid phases and the molten egg of corn alcohol that dry protein content is 79% is obtained after washing, dehydrating Content of prolamine in white product, albumen is more than 85.60%, wherein molten containing 87.5% alpha-alcohol soluble protein, 3.1% β -ol Albumen and the 5.1% molten albumen of γ -ol, fat and ash content contents on dry basis are respectively 1.16% and 0.51%.In addition, the second filter cake The product of dry protein content 49% is obtained after drying.

Embodiment 22

Aqueous 31.0% wet vinasse (WDG) are added water into regulation moisture to 80%, using colloid mill (MagicLab, IKA companies) (about 30 μm of abrasive disk space) is crushed, squeeze into enzymolysis and piece-rate system：Material initially enters the first enzymatic vessel, PH3.5, alpha amylase (the Liquozyme SCDS, Novi's letter limited public affairs of enzyme preparation that starch contained therein weight 5wt% is added at 30 DEG C Department), 3wt% carbohydrase (Spirizyme Ultra, letter enzyme preparation Co., Ltd of Novi), 2wt% Pullulanase (letter enzyme preparation Co., Ltd of Promozyme D2 Novi), 10% composite alpha-amylase (Novozyme NS50013, Novi's letter enzyme system Agent Co., Ltd), 2mM magnesium ions and 2mM calcium ions react 1 hour；Then adjust pH to 5.6 and added at 50 DEG C contained fine The composite fibre hydrolase (GC 518, Jie Nengke bioengineering Co., Ltd) for tieing up weight 10% reacts 5 hours；Material is squeezed into Second enzymatic vessel, adds contained protein by weight 0.03wt% neutral proteinase (SUKAPro NE, Su Kehan at pH8,20 DEG C Bioengineering limited company) and 20mM mercaptoethanols react 5 hours, then adjust pH to 10 maintain 0.2 hour.Use 1 μm Aperture membrane filtration obtains the first permeate and trapped fluid, and regulation trapped fluid moisture squeezes into the 3rd enzymatic vessel to 80%, adjusts pH Add materials contained protein by weight 0.02wt% alkali protease (limited public affairs of 2709, Pang Bo bioengineering to 10, after 55 DEG C Department) 0.5 hour is incubated, the second filtrate and trapped fluid are then obtained by 20 μm of filter screen, egg is obtained after filtrate is dehydrated Content of prolamine in the zeins product that white contents on dry basis is 94.8%, albumen is more than 93.7%, wherein containing 81.1% alpha-alcohol soluble protein, the 17.6% molten albumen of β -ol and the 1.3% molten albumen of γ -ol, fat and ash content contents on dry basis Respectively 2.48% and 0.70%.In addition, the second trapped fluid obtains the product of dry protein content 55% after drying.

Comparative example

By aqueous 9.0% distiller's dried grain (DDG), using airslide disintegrating mill, (FQS15 types, Shanghai causes triumphant powder machinery manufacture to have Limit company) powder particle diameter extracts 30min to adding 70% ethanol and 3.5% sodium hydroxide solution after about 100 μm at 70 DEG C.Its In, ethanol solution accounts for the 83% of feed liquid gross weight.Then use centrifuge (ALLEGRA 30R, the limited public affairs of U.S.'s Beckman Kurt Department) clear liquid (2000g, 10min) is isolated, by clear liquid using the membrane filtration that aperture is 1 micron, filtrate is again by retaining molecule The filter membrane measured as 10kDa is done after being concentrated in vacuum drying chamber (DZF-6210, Shanghai Yiheng Scientific Instruments Co., Ltd) Crushing is the product that 150 destination protein contents on dry basis are 86.4% after dry.

Table 2：Embodiment 17-22 product form

Granularmetric analysis

The measure of particle diameter uses the laser particle analyzers of Mastersizer 3000 of Malvern Instrument companies of Britain Test, test adopts water as medium, takes a certain amount of product in medium, mechanical agitation makes it be well dispersed in medium, grain Degree instrument, which is determined, can provide average grain diameter.

The measure of alcohol soluble protein product colour

Take the use embodiment 17-22 of 150 mesh and the alcohol soluble protein sample of comparative example method preparation and commercially available alcohol molten Protein sample (dry protein content 89% is purchased from Rong Ning trade Co., Ltds of Zhuhai City).In color difference meter (CR2400, Japanese Ke Buddhist nun's card Minolta color difference meter) on record powder surface color L (brightness value), a (red scale value) and b (yellow value degree) value.

From table 3 below, compared with alcohol soluble protein sample commercially available and prepared by comparative example method, in embodiment 17-22 The yellow value degree (b) of alcohol soluble protein product have obvious reduction, can at least reduce about 20%, maximum can reduce about 78%.Illustrate this Invented technology also has good decolorizing effect.Product of the present invention is as dispensing to original application system (such as food or medicine) face The influence of color is lower.

Table 3：LAB colour measurement results

Alcohol soluble protein aerogenesis taste subjective appreciation

Alcohol soluble protein sample and commercially available alcohol soluble protein sample (egg prepared by Example 17-22 and comparative example method White contents on dry basis 89%, is purchased from Rong Ning trade Co., Ltds of Zhuhai City) each 25g.In 20~22 DEG C of room temperature, relative humidity is maintained at 55% -65% or so.Sample carries out zeins characteristic odor evaluation and test under yellow light sources irradiation.According to odour intensity It is divided into 7 grades, respectively 1- does not have, 2- does not have substantially, 3- is not more obvious, and 4- is general, and 5- is more obvious, substantially, 7- is very by 6- Substantially.Test number is 25 people.As a result rounded for average value.

From table 4 below, the characteristic odor of the alcohol soluble protein product in embodiment 17-22 is that general (4) arrive basic Do not have between (2), technique has the effect of deodorization concurrently in itself.Product of the present invention as dispensing to original application system (such as food or Medicine) smell influence it is lower.

Table 4：Smell results of sensory evaluation

Alcohol soluble protein product filming performance compares

Alcohol soluble protein sample and commercially available alcohol soluble protein sample that in Example 17-22 prepared by sample, comparative example method Product carry out contrast test.A certain amount of above-mentioned alcohol soluble protein sample is weighed, 75% ethanol solution is added, is configured to 10wt% albumen Matter solution, is uniformly mixed, after filter paper or membrane filtration, is separately added into 20% glycerine and PEG-4000 (with protein Amount ratio), 20min is stirred, is put into after 80 DEG C of water bath with thermostatic control heating stirring 15min and takes out, the albumen of certain volume is noted into film liquid Film is taken off after entering film forming pallet, 50 DEG C of air heat drying film forming, 2h, is balanced in 45% relative humidity and room temperature environment after 24h Determine mechanical performance.The homogeneous maize alcohol-soluble protein film of quality is cut into 15mm × 50mm size, with reference to GB/T6672- 2001 methods determine its thickness symmetrical 5 points measurement thickness of selection on albumen membrane sample, average.The mechanical performance of film It is 1mm/s to determine draw speed when is determined with Texture instrument (TA-XT2i, Stable Micro Systems companies of Britain), film Effective measured length is 80mm.Power when tensile strength (TS) is broken for film in drawing process on unit cross section, calculation formula It is as follows：

In formula：TS is tensile strength, MPa；F is maximum pull (N)；δ is the thickness (mm) of film, mm；W is the width of membrane sample Spend (W=15mm).

As seen from Figure 5, compared with commercial samples, the alcohol soluble protein product in embodiment 17-22 be equally respectively provided with directly into Film.Content of prolamine is linear in the tensile strength and sample of protein film, the good (R of fitting²=0.8766).On an equal basis Under content of prolamine, the tensile strength of sample (such as embodiment 22) protein film of the invention can also slightly be better than the commercially available jade of reference The film that rice alcohol soluble protein is made.

Alcohol soluble protein microballoon stability compares

For the micro-capsule stability of alcohol soluble protein contained by sample in comparing embodiment 17-22, by the reality in addition to embodiment 18 Apply one in a sample (such as embodiment 17) and comparative example reuse 80% ethanol solution and extracts 30min at 65 DEG C, Obtain the sample that alcohol soluble protein contents on dry basis is more than 85%.By the sample of embodiment 2, commercial samples (dry protein content 89%, It is purchased from Rong Ning trade Co., Ltds of Zhuhai City) together with freshly prepd embodiment 17, embodiment 19-22, comparative example, point Do not take the appropriate volume fraction that is dissolved in 80% ethanol solution, to prepare alcohol soluble protein ethanol solution, mass fraction is controlled For 3%.In the case where high-speed stirred speed is 12000rpm, above-mentioned zein ethanol solutions is injected in appropriate high purity water, pass through film Separation controls the alcohol soluble protein mass fraction of final system to obtain alcohol soluble protein microspheres solution for 1%.By depressurizing low-temperature evaporation It is 3% or so to make alcohol soluble protein quality in the solution.By the microspheres solution as in graduated vessels and under refrigerated condition (4 DEG C) Preserve, the apparent coagulation time (i.e. beds of precipitation volume is more than the 5% of total solution volume) occurs for record during storage.

From table 5 below, alcohol soluble protein product (embodiment 18) of the invention or the secondary alcohol extracted again from product Microspheres solution prepared by molten protein product (embodiment 17) has good stability.Simultaneously with an existing solvent extraction process Alcohol soluble protein product (such as comparative example and commercially available prepared by (directly being extracted from raw material using ethanol or other organic solvents) Product) compare, with longer stabilization time.

Table 5：Microballoon stability

	Embodiment 17	Embodiment 18	Comparative example	Commercially available zeins
					The coagulation time (my god)	112	105	90	92

In view of exemplary method and apparatus, with reference to the flow chart of each figure, be better understood with can be according to the master of the disclosure The method that body is implemented, but in order to simplify the purpose of explanation, method is shown and described with a series of block diagrams, should and it can manage Solution, because the order that some block diagrams may be different from what is described and describe occurs and/or occurred with other block diagrams simultaneously The theme of protection is asked not limited by the order of block diagram.In addition, the block diagram of not all explanation is all the institute in implementation Need.

Claims (6)

1. a kind of protein product, the protein product includes alcohol soluble protein and carbohydrate, it is characterised in that the molten egg of alcohol White more than the 70wt% for accounting for albumen butt；Meanwhile, alpha-alcohol soluble protein accounts for more than the 75wt% of the alcohol soluble protein, the molten albumen of β -ol Below the 20wt% of the alcohol soluble protein is accounted for, the molten albumen of γ -ol accounts for below the 6wt% of the alcohol soluble protein.

2. protein product as claimed in claim 1, it is characterised in that the alcohol soluble protein accounts for the 74wt% of the albumen butt The above, more preferably preferably more than 80wt%, more than 85wt%, even more preferably further preferred more than 90wt%, 95wt% Above, most preferably more than 97wt%.

3. protein product as claimed in claim 1 or 2, it is characterised in that the alpha-alcohol soluble protein accounts for the alcohol soluble protein More than 77wt%, preferably more than 85wt%, more preferably more than 90wt%, further preferred more than 95wt%, most preferably 100wt%.

4. the protein product as any one of claim 1-3, it is characterised in that it is molten that the molten albumen of β -ol accounts for the alcohol Below the 10wt% of albumen, preferably below 5wt%, more preferably below 3wt%, further preferred below 2wt%, most preferably 0%.

5. the protein product as any one of claim 1-4, it is characterised in that the molten albumen of γ -ol accounts for the alcohol Below the 10wt% of molten albumen, preferably below 5wt%, more preferably below 2wt%, most preferably 0%.

6. the protein product as any one of claim 1-5, it is characterised in that the albumen butt accounts for the albumen production More than the 85wt% of product, preferably below 90wt%, more preferably more than 94wt%, most preferably more than 99wt%.