CN112239772A - Corn polypeptide powder and method for extracting corn polypeptide by ultrasonic-assisted enzyme method - Google Patents
Corn polypeptide powder and method for extracting corn polypeptide by ultrasonic-assisted enzyme method Download PDFInfo
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Abstract
The invention relates to a corn polypeptide extraction process, in particular to corn polypeptide powder and a method for extracting corn polypeptide by an ultrasonic-assisted enzyme method, wherein the method comprises the following steps: 1) contacting corn fermented mash with saccharifying enzyme for first enzymolysis, and performing solid-liquid separation on a first enzymolysis product to obtain a solid-phase enzymolysis product; 2) mixing the solid phase enzymolysis product with water, contacting with protease for second enzymolysis, and performing second solid-liquid separation on the second enzymolysis product to obtain hydrolysate rich in polypeptide; 3) heating the hydrolysate to inactivate enzyme, cooling, concentrating, and drying to obtain corn polypeptide powder; the first enzymolysis and/or the second enzymolysis is carried out under the condition of ultrasound. The method adopts ultrasonic method and carbohydrase for auxiliary extraction, promotes the pulverization and liquefaction of alcohol soluble protein, improves the activity of protease molecules, increases the contact probability of alcohol soluble protein and protease active sites, improves the utilization rate of protease, remarkably improves the extraction efficiency, and shortens the extraction time.
Description
Technical Field
The invention relates to a corn polypeptide extraction process, in particular to corn polypeptide powder and a method for extracting corn polypeptide by an ultrasonic-assisted enzyme method.
Background
In recent years, the yield of fuel ethanol as a renewable bio-energy source has been increased dramatically with the increasing environmental problems of coal and petroleum energy crisis, air pollution, global warming, and the like. The biofuel ethanol process in China mainly takes corn as a raw material for production, and the ethanol is produced through liquefaction, synchronous saccharification and fermentation and distillation, and simultaneously, a large amount of vinasse mash is produced. The vinasse mash contains a large amount of crude protein, rich amino acid, microorganism and various trace elements, and if the vinasse mash is not properly treated, huge resource waste and serious environmental pollution can be caused, and if the vinasse mash is properly treated, the problem of environmental pollution can be solved, a byproduct with higher economic value can be obtained, and the economic benefit of the corn fuel ethanol is improved.
Zein (zein) is an important protein in zein and accounts for 50-60% of the total protein. The alcohol soluble protein is a protein with strong hydrophobicity, is insoluble in water, is soluble in 60-95% alcohol aqueous solution, and can also be soluble in strong base solution with pH value being more than 11, high concentration urea or anion active agent (such as sodium dodecyl sulfate SDS and the like), which is determined by the special amino acid composition, and contains a large amount of hydrophobic amino acid and sulfur-containing amino acid, such as glutamic acid, leucine, proline and alanine, but lacks acidic and basic amino acids such as lysine, aspartic acid and tryptophan, and can not ensure nitrogen balance when being used as food, but the unbalanced amino acid composition enables zein to be a source of functional active peptide.
The method for obtaining various physiologically active peptides by carrying out enzymolysis on zein by adopting protease is an important research means for improving the comprehensive utilization rate of zein. But zein has a unique and complex internal structure like other natural proteins, alpha-spirochetes are formed by the hydrogen bonding of hydroxyl groups and imino groups on the polypeptide main chain, polypeptide chains are combined with each other through secondary bonds to form a complex quaternary structure, and natural zein molecules can be further polymerized into polymers, so the zein has a complex molecular structure and high compactness, and has strong resistance to enzymatic attack of protease. In addition, the zein is difficult to dissolve in water, and is suspended in an aqueous solution with low probability of contacting with protease. Therefore, the traditional protease enzymolysis method has the problems of low product yield, long enzymolysis time, low protease utilization rate and the like.
Disclosure of Invention
The invention aims to overcome the problems of low product yield, long enzymolysis time, low protease utilization rate and the like of the traditional protease method in the prior art, and provides a method for extracting corn polypeptide by using an ultrasonic-assisted enzyme method and corn polypeptide powder extracted by using the method.
In order to achieve the above objects, the present invention provides a method for extracting corn polypeptide, comprising the steps of:
step 1: contacting the corn fermented mash with saccharifying enzyme to carry out first enzymolysis on the corn fermented mash, and carrying out first solid-liquid separation on an obtained first enzymolysis product to obtain a solid-phase enzymolysis product;
step 2: mixing the obtained solid-phase enzymolysis product with water, contacting with protease to carry out second enzymolysis on the solid-phase enzymolysis product, and carrying out second solid-liquid separation on the obtained second enzymolysis product to obtain hydrolysate rich in polypeptide;
and step 3: heating the hydrolysate to inactivate enzyme, cooling, concentrating, and drying to obtain corn polypeptide powder;
wherein the first enzymatic hydrolysis and/or the second enzymatic hydrolysis are/is carried out under the condition of ultrasound.
In another aspect, the present invention provides a corn polypeptide flour prepared by the method as described above.
The method for extracting the corn polypeptide by the ultrasonic-assisted enzymatic method provided by the invention has the following advantages:
(1) according to the invention, firstly, the corn fermented mash is treated by adopting the glycoenzyme method, so that the outer layer of protein substances in the fermented mash is effectively removed, the hydrolysate rich in monosaccharide is obtained, the structure of the corn is further opened, the protein in the solid is more easily contacted, on the basis of the first hydrolysis step, the protease is more easily contacted with the hydrolysate, the enzymolysis efficiency is further improved, and the hydrolysate rich in polypeptide is obtained.
(2) The invention adopts ultrasonic auxiliary treatment in the enzymolysis process, not only modifies the structures of substrate molecules and enzyme molecules, but also increases the contact probability of the substrate and the enzyme active site, promotes the occurrence of enzymolysis reaction, effectively improves the utilization rate of enzyme, has high product yield, obviously shortens the enzymolysis time, and has very high economic benefit and social benefit.
(3) The invention has the advantages of safety, environmental protection, green and the like, improves the added value of the corn fermented mash, obtains the corn polypeptide finished product with high purity, can be completely dissolved in water, enlarges the application range of the zein, and has wide application prospect.
(4) The invention reduces resource waste, improves economic benefit, truly realizes the value of zein, provides certain theoretical guidance for future industrial production, and has important significance.
Drawings
FIG. 1 is a process flow diagram of a specific ultrasonic-assisted enzymatic extraction of maize polypeptides of the present invention.
Detailed Description
The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value, and such ranges or values should be understood to encompass values close to those ranges or values. For ranges of values, between the endpoints of each of the ranges and the individual points, and between the individual points may be combined with each other to give one or more new ranges of values, and these ranges of values should be considered as specifically disclosed herein.
In a first aspect, the present invention provides a method for extracting maize polypeptides, comprising the steps of:
step 1: contacting the corn fermented mash with saccharifying enzyme to carry out first enzymolysis on the corn fermented mash, and carrying out first solid-liquid separation on an obtained first enzymolysis product to obtain a solid-phase enzymolysis product;
step 2: mixing the obtained solid-phase enzymolysis product with water, contacting with protease to carry out second enzymolysis on the solid-phase enzymolysis product, and carrying out second solid-liquid separation on the obtained second enzymolysis product to obtain hydrolysate rich in polypeptide;
and step 3: heating the hydrolysate to inactivate enzyme, cooling, concentrating, and drying to obtain corn polypeptide powder;
wherein the first enzymatic hydrolysis and/or the second enzymatic hydrolysis are/is carried out under the condition of ultrasound.
According to the invention, the corn fermented mash is a byproduct in the alcohol production process by using corn as a raw material, specifically, the corn raw material is liquefied and saccharified synchronously, then a strain is inoculated for alcohol fermentation, alcohol is distilled after the fermentation is finished, and the residual liquid is the corn fermented mash which contains a large amount of crude protein, abundant amino acid, microorganism and various trace elements. The source of the corn beer is not particularly limited and may be obtained commercially or prepared by itself under conditions that provide corn alcohol production capacity.
According to a preferred embodiment of the invention, the corn beer contains 3-4 wt.% crude protein.
According to the present invention, the object of the present invention can be achieved by introducing ultrasound in any one of the first enzymatic hydrolysis step and the second enzymatic hydrolysis step, but more preferably, both the first enzymatic hydrolysis step and the second enzymatic hydrolysis step are performed under the condition of ultrasound.
According to the present invention, preferably, the first enzymatic hydrolysis is performed under the condition of ultrasound, which can be changed within a wide range, but in order to further enhance the effect of the present invention, the power of the ultrasound is 100-.
According to the present invention, the conditions of the first enzymatic hydrolysis can be selected within a wide range, and preferably, in order to further enhance the effect of the present invention, the conditions of the first enzymatic hydrolysis include a pH of 4 to 5.5, for example, 4, 4.3, 4.5, 4.8, 5, 5.3, 5.5, preferably 4.8 to 5; the temperature is 40 to 60 ℃, for example, 40 ℃, 45 ℃, 47 ℃, 51 ℃, 53 ℃, 55 ℃, 60 ℃, preferably 45 to 55 ℃, and the total time is 6 to 48 hours, for example, 6 hours, 10 hours, 15 hours, 20 hours, 25 hours, 30 hours, 35 hours, 40 hours, 45 hours, 48 hours, preferably 20 to 30 hours.
According to the present invention, the first enzymatic hydrolysis may be performed under the ultrasonic condition in the whole process, or may be performed under the ultrasonic condition in a certain time period, for example, in the initial stage, in the intermediate stage, or in the end stage. According to a preferred embodiment of the present invention, the initial stage of the first enzymatic hydrolysis is performed under the condition of ultrasound, and the ultrasound time is preferably within 3h, for example, 0.1h, 0.2h, 0.4h, 0.6h, 0.8h, 1h, 1.2h, 1.4h, 1.6h, 1.8h, 2h, 2.2h, 2.4h, 2.6h, 2.8h, 3h, more preferably 1.5-2.5 h.
According to a more preferred embodiment of the invention, the mixture of corn fermented mash and saccharifying enzyme is subjected to ultrasonic enzymolysis, and then stirred for enzymolysis until the enzymolysis is finished.
Wherein the rotation speed of the stirring is not particularly limited, and is preferably 200-300 rpm.
According to the present invention, the amount of the saccharifying enzyme can be selected from a wide range, and in order to further improve the effect of the present invention, it is preferable that the saccharifying enzyme is added in an amount of 0.005 to 0.15 parts by weight, for example, 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.11, 0.12, 0.13, 0.14, 0.15, preferably 0.08 to 0.12 parts by weight, based on 100 parts by weight of the corn mash on a dry weight basis.
According to the present invention, in order to further improve the effect of the present invention, in step 1, the first enzymatic hydrolysis is further carried out in the presence of cellulase and hemicellulase, and the cellulase may be added in an amount of not more than 0.6 parts by weight, for example, 0.1 parts by weight, 0.2 parts by weight, 0.3 parts by weight, 0.4 parts by weight, 0.5 parts by weight, 0.6 parts by weight, and preferably 0.4 to 0.6 parts by weight, and hemicellulose may be added in an amount of not more than 0.3 parts by weight, for example, 0.1 parts by weight, 0.2 parts by weight, 0.3 parts by weight, and preferably 0.1 to 0.3 parts by weight, relative to 100 parts by weight of the corn beer on a dry basis.
It should be noted that the dry weight of the corn beer as described above refers to the weight of dry matter obtained by drying the corn beer at 40-60 ℃ to a constant weight.
According to the present invention, the first solid-liquid separation method may be any conventional method for separating a solid phase and a liquid phase in a mixed liquid, and may be, for example, filtration, centrifugation or the like. According to a preferred embodiment of the invention, the first solid-liquid separation is centrifugation. Wherein, the centrifugation conditions can be selected in a wide range, and preferably, the centrifugation conditions are as follows: the rotation speed is 3000-10000rpm, for example, 3000rpm, 3500rpm, 4000rpm, 4500rpm, 5000rpm, 6000rpm, 7000rpm, 8000rpm, 9000rpm, 10000rpm, preferably 3000-8000 rpm; the time is 5-30min, for example, 5min, 10min, 15min, 20min, 25min, 30min, preferably 15-25 min.
According to the present invention, in order to improve the purity of the finally obtained corn polypeptide, it is preferable that the solid-liquid separation is followed by washing the obtained solid with water several times until there is no free residual sugar on the surface. Wherein the water may be deionized water, pure water, etc.
In step 2, the amount of water used is not particularly limited, but preferably the amount of water is such that the dry solids content of the material after mixing the solid phase enzymatic hydrolysate with water is 25 to 35 wt%, preferably 27 to 33 wt%, for example 27 wt%, 28 wt%, 29 wt%, 30 wt%, 31 wt%, 32 wt%, 33 wt%. Wherein the water may be deionized water, pure water, etc. The dry solid content is the weight of dry matter obtained by drying the solid phase enzymolysis product to constant weight under room temperature.
According to the present invention, the protease may be a protease used in a conventional protease enzymatic method, for example, may include, but is not limited to, at least one of an alkaline protease, a neutral protease, and an acidic protease; preferably at least one of flavourzyme, cotinase, bromelain, papain and trypsin.
According to the present invention, the amount of the protease is not particularly limited, and preferably is not more than 20 parts by weight, for example, 0.5 parts by weight, 1 part by weight, 5 parts by weight, 7 parts by weight, 9 parts by weight, 11 parts by weight, 13 parts by weight, 15 parts by weight, 17 parts by weight, or 20 parts by weight, relative to 100 parts by weight of the solid-phase enzymatic product on a dry weight basis.
According to the invention, preferably, the second enzymolysis is carried out under the condition of ultrasound, the condition of ultrasound can be changed in a wide range, but in order to further improve the effect of the invention, the power of ultrasound is 100-300W, for example, 100W, 110W, 120W, 130W, 140W, 160W, 180W, 200W, 220W, 240W, 260W, 280W, 300W, preferably 110-140W.
According to the invention, the conditions for the second enzymatic hydrolysis can be selected within a wide range, preferably at a temperature of 40-60 ℃, for example 40 ℃, 45 ℃, 47 ℃, 51 ℃, 53 ℃, 55 ℃, 60 ℃, preferably 45-55 ℃, for a total time of 2-48 hours, for example 2 hours, 6 hours, 10 hours, 15 hours, 20 hours, 25 hours, 30 hours, 35 hours, 40 hours, 45 hours, 48 hours, preferably 10-15 hours.
According to the present invention, the pH value of the second enzymatic hydrolysis process can be specifically selected according to the kind of the protease to be added, and is preferably controlled within the optimum pH value range of the protease to be added.
According to the present invention, the second enzymatic hydrolysis may be performed under the ultrasonic condition in the whole process, or may be performed under the ultrasonic condition in a certain time period, for example, in the initial stage, in the intermediate stage, or in the end stage. According to a preferred embodiment of the present invention, the initial stage of the second enzymatic hydrolysis is performed under the condition of ultrasound, and the ultrasound time is preferably within 3h, for example, 0.1h, 0.2h, 0.4h, 0.6h, 0.8h, 1h, 1.2h, 1.4h, 1.6h, 1.8h, 2h, 2.2h, 2.4h, 2.6h, 2.8h, 3h, more preferably 0.3-0.8 h.
According to a more preferred embodiment of the present invention, the second enzymolysis method comprises subjecting the mixture of the solid phase enzymolysis product, water and protease to ultrasonic enzymolysis, and then stirring for enzymolysis.
Wherein the rotation speed of the stirring is not particularly limited, and is preferably 200-300 rpm.
According to the present invention, the second solid-liquid separation method may be any conventional method for separating a solid phase and a liquid phase in a mixed liquid, and may be, for example, filtration, centrifugation or the like. According to a preferred embodiment of the invention, the second solid-liquid separation is centrifugation. Wherein, the centrifugation conditions can be selected in a wide range, and preferably, the centrifugation conditions are as follows: the rotation speed is 3000-10000rpm, for example, 3000rpm, 3500rpm, 4000rpm, 4500rpm, 5000rpm, 6000rpm, 7000rpm, 8000rpm, 9000rpm, 10000rpm, preferably 3000-8000 rpm; the time is 5-30min, for example, 5min, 10min, 15min, 20min, 25min, 30min, preferably 15-25 min.
According to the present invention, in step 3, the temperature for enzyme inactivation is not particularly limited as long as the added enzyme can be effectively inactivated, and for example, the enzyme in the system can be inactivated by heating at 90-110 ℃ for 5-15 min.
According to the present invention, in step 3, the concentration method may also be a method conventional in the art, for example, concentration by heating, concentration under reduced pressure, and the like.
According to the present invention, in step 3, the drying method may be a method conventional in the art, and for example, may be freeze drying, spray drying or vacuum heat drying.
According to a preferred embodiment of the present invention, as shown in FIG. 1, the method for extracting corn polypeptide comprises:
(1) adjusting the pH value of the corn fermented mash to 4.8-5, adding diastase, cellulase and hemicellulase, wherein the addition amount of the diastase is 0.08-0.12% of the dry weight of solids, the addition amount of the cellulase is 0.4-0.6% of the dry weight of the solids, the addition amount of the hemicellulase is 0.1-0.3% of the dry weight of the solids, performing enzymolysis for 20-25h under the condition of 200-300rpm after performing ultrasonic treatment for 1.5-2.5h under 110-140W, and keeping the temperature at 45-55 ℃ in the whole enzymatic hydrolysis process.
(2) After enzymatic hydrolysis, solid-liquid separation, washing the solid with pure water several times until there is no free residual sugar on the surface.
(3) Mixing the solid and pure water until the dry matter concentration is 27-33 wt%, adjusting the pH, adding alkaline protease, wherein the addition amount of the alkaline protease is 5-15% of the dry weight of the solid, firstly performing ultrasonic treatment at 110-140W for 0.3-0.8h, then performing enzymolysis at 200-300rpm for 10-15h, and keeping the temperature at 45-55 ℃ in the whole enzymatic hydrolysis process.
(4) After enzymatic hydrolysis, performing solid-liquid separation, rapidly heating the hydrolysate to 90-110 ℃, keeping for 5-15min, inactivating enzyme, cooling, concentrating under reduced pressure, and freeze-drying to obtain corn polypeptide powder.
In a second aspect, the present invention provides corn polypeptide meal prepared by the method described above.
The present invention will be described in detail below by way of examples. In the following examples of the present invention,
the corn beer stock from the corn fermentation line contained 3.5 wt% crude protein.
The concentrations of glucose, xylose and ethanol are detected by high performance liquid chromatography.
The content of crude protein and polypeptide is detected by adopting a Kjeldahl method (GB/T5009.5-2010) and calculated to obtain the polypeptide.
The polypeptide extraction rate is the mass (g) of the polypeptide after extraction/the total protein content (g) in the fermented mash.
The alkaline protease is purchased from Shanghai-derived leaf Biotech, Inc., cat # 9014-01-1.
Example 1
This example illustrates the method of extracting corn polypeptides from corn beer provided by the present invention
(1) Adjusting the pH value of 100g of corn fermented mash to 4.8-5, adding diastase, cellulase and hemicellulase, wherein the addition amount of the diastase is 0.1% of the dry weight of solids, the addition amount of the cellulase is 0.6% of the dry weight of the solids, the addition amount of the hemicellulase is 0.1% of the dry weight of the solids, performing enzymolysis for 22h under the condition of 250rpm after performing ultrasonic treatment for 2h under 120W, and keeping the temperature at 50 ℃ in the whole enzymatic hydrolysis process.
(2) After the enzymatic hydrolysis, solid-liquid separation was carried out, the solid was washed with pure water several times until there was no free residual sugar on the surface, the washing water was combined with the liquid phase obtained in the step (1) and the solid-liquid separation, and the amount of glucose in the liquid phase was measured, and the results are shown in Table 1.
(3) Mixing the solid with pure water until the dry matter concentration is 30 wt%, adjusting pH, adding alkaline protease, wherein the addition amount of the alkaline protease is 5% of the dry matter weight, performing ultrasonic treatment at 120W for 0.5h, performing enzymolysis at 250rpm for 12h, and keeping the temperature at 50 ℃ in the whole enzymatic hydrolysis process.
(4) After enzyme hydrolysis, carrying out solid-liquid separation, rapidly heating the hydrolysate to 100 ℃ for 10min, inactivating enzyme, cooling, concentrating under reduced pressure, and freeze-drying to obtain corn polypeptide powder, wherein the quality and extraction rate of the corn polypeptide powder are shown in the table.
Example 2
This example illustrates the method of extracting corn polypeptides from corn beer provided by the present invention
(1) Adjusting the pH value of 100g of corn fermented mash to 4.8-5, adding diastase, cellulase and hemicellulase, wherein the addition amount of the diastase is 0.08% of the dry weight of solids, the addition amount of the cellulase is 0.4% of the dry weight of the solids, the addition amount of the hemicellulase is 0.3% of the dry weight of the solids, performing enzymolysis for 25h under the condition of 200rpm after performing ultrasonic treatment for 1.5h under 110W, and keeping the temperature at 55 ℃ in the whole enzymatic hydrolysis process.
(2) After the enzymatic hydrolysis, solid-liquid separation was carried out, the solid was washed with pure water several times until there was no free residual sugar on the surface, the washing water was combined with the liquid phase obtained in the step (1) and the solid-liquid separation, and the amount of glucose in the liquid phase was measured, and the results are shown in Table 1.
(3) Mixing the solid with pure water until the dry matter concentration is 27 wt%, adjusting pH, adding alkaline protease, wherein the addition amount of the alkaline protease is 10% of the dry matter weight of the solid, performing ultrasonic treatment at 110W for 0.3h, performing enzymolysis at 200rpm for 15h, and keeping the temperature at 45 ℃ in the whole enzymatic hydrolysis process.
(4) After enzymatic hydrolysis, performing solid-liquid separation, rapidly heating the hydrolysate to 110 ℃ for 5min, inactivating enzyme, cooling, concentrating under reduced pressure, and freeze-drying to obtain corn polypeptide powder with quality and extraction rate shown in the table.
Example 3
This example illustrates the method of extracting corn polypeptides from corn beer provided by the present invention
(1) Adjusting the pH value of 100g of corn fermented mash to 4.8-5, adding diastase, cellulase and hemicellulase, wherein the addition amount of the diastase is 0.12% of the dry weight of solids, the addition amount of the cellulase is 0.5% of the dry weight of the solids, the addition amount of the hemicellulase is 0.2% of the dry weight of the solids, performing enzymolysis for 20h under the condition of 300rpm after performing ultrasonic treatment for 2.5h under 130W, and keeping the temperature at 45 ℃ in the whole enzymatic hydrolysis process.
(2) After the enzymatic hydrolysis, solid-liquid separation was carried out, the solid was washed with pure water several times until there was no free residual sugar on the surface, the washing water was combined with the liquid phase obtained in the step (1) and the solid-liquid separation, and the amount of glucose in the liquid phase was measured, and the results are shown in Table 1.
(3) Mixing the solid with pure water until the dry matter concentration is 33 wt%, adjusting pH, adding alkaline protease, wherein the addition amount of the alkaline protease is 15% of the dry matter weight, performing ultrasonic treatment at 130W for 0.8h, performing enzymolysis at 300rpm for 10h, and keeping the temperature at 55 ℃ in the whole enzymatic hydrolysis process.
(4) After enzymatic hydrolysis, performing solid-liquid separation, rapidly heating the hydrolysate to 90 ℃ for 5min, inactivating enzyme, cooling, concentrating under reduced pressure, and freeze-drying to obtain corn polypeptide powder with quality and extraction rate shown in the table.
Example 4
This example illustrates the method of extracting corn polypeptides from corn beer provided by the present invention
Zein was extracted from corn beer according to the method of example 1, except that the enzymatic hydrolysis of step (1) was not carried out under ultrasound.
Example 5
This example illustrates the method of extracting corn polypeptides from corn beer provided by the present invention
Zein was extracted from corn beer according to the method of example 1, except that the enzymatic hydrolysis of step (3) was not performed under ultrasound.
Example 6
This example illustrates the method of extracting corn polypeptides from corn beer provided by the present invention
Corn protein was extracted from corn beer according to the method of example 1, except that in both step (1) and step (3), the corn protein was enzymatically hydrolyzed under agitation and then under ultrasound.
Example 7
This example illustrates the method of extracting corn polypeptides from corn beer provided by the present invention
Corn protein was extracted from corn beer according to the method of example 1, except that cellulase and hemicellulase were not added during the enzymatic hydrolysis of step (1), and the amount of amylosaccharifying enzyme was increased accordingly.
Example 8
This example illustrates the method of extracting corn polypeptides from corn beer provided by the present invention
Corn protein was extracted from corn beer according to the method of example 1 except that in step (3), the alkaline protease was replaced with an equal amount of the acidic protease.
Comparative example 1
This comparative example serves to illustrate a reference method for extracting corn polypeptide from corn beer
Zein was extracted from corn beer according to the method of example 1, except that the enzymatic hydrolysis in step (1) and step (3) was not carried out under sonication.
Comparative example 2
This comparative example serves to illustrate a reference method for extracting corn polypeptide from corn beer
Corn protein was extracted from corn beer according to the method of example 1, except that step (1) was omitted.
TABLE 1
The results in table 1 show that the ultrasonic method and the carbohydrase-assisted extraction are adopted in the invention, so that the extraction efficiency is remarkably improved, the extraction time is shortened, and meanwhile, sugar solution with higher glucose content can be obtained.
The preferred embodiments of the present invention have been described above in detail, but the present invention is not limited thereto. Within the scope of the technical idea of the invention, many simple modifications can be made to the technical solution of the invention, including combinations of various technical features in any other suitable way, and these simple modifications and combinations should also be regarded as the disclosure of the invention, and all fall within the scope of the invention.
Claims (10)
1. A method for extracting corn polypeptide is characterized by comprising the following steps:
step 1: contacting the corn fermented mash with saccharifying enzyme to carry out first enzymolysis on the corn fermented mash, and carrying out first solid-liquid separation on an obtained first enzymolysis product to obtain a solid-phase enzymolysis product;
step 2: mixing the obtained solid-phase enzymolysis product with water, contacting with protease to carry out second enzymolysis on the solid-phase enzymolysis product, and carrying out second solid-liquid separation on the obtained second enzymolysis product to obtain hydrolysate rich in polypeptide;
and step 3: heating the hydrolysate to inactivate enzyme, cooling, concentrating, and drying to obtain corn polypeptide powder;
wherein the first enzymatic hydrolysis and/or the second enzymatic hydrolysis are/is carried out under the condition of ultrasound.
2. The method of claim 1, wherein the conditions of the ultrasound comprise: the ultrasonic power is 100-300W, and the ultrasonic time is within 3 h.
3. The method of claim 1 or 2, wherein the first enzymatic hydrolysis and the second enzymatic hydrolysis are performed under conditions of sonication.
4. The method according to any one of claims 1 to 3, wherein in the step 1, the conditions of the first enzymolysis comprise that the mixture of the solid phase enzymolysis product and the protease is subjected to ultrasonic enzymolysis, and then is subjected to stirring enzymolysis;
preferably, the pH value of the first enzymolysis is 4-5.5, the temperature is 40-60 ℃, and the total time is 6-48 hours;
preferably, the saccharifying enzyme is added in an amount of 0.005-0.15 parts by weight based on 100 parts by weight of the corn beer on a dry basis.
5. The method according to any one of claims 1-4, wherein in step 1, the first enzymatic hydrolysis is further carried out in the presence of cellulase enzymes and hemicellulase enzymes, the cellulase enzymes being added in an amount of not more than 0.6 parts by weight and the hemicellulose being added in an amount of not more than 0.3 parts by weight, relative to 100 parts by weight of the corn beer on a dry basis.
6. A process according to any one of claims 1 to 5, wherein in step 2, the solids content of the solid phase enzymatic hydrolysate after mixing with water is 25 to 35 wt%, preferably 27 to 33 wt%.
7. The method according to any one of claims 1 to 6, wherein in step 2, the protease is selected from at least one of an alkaline protease, a neutral protease and an acidic protease;
preferably, in step 2, the protease is at least one selected from flavourzyme, complex protease, bromelain, papain and trypsin.
8. The method according to any one of claims 1 to 7, wherein in the step 2, the second enzymolysis condition comprises subjecting the mixture of the solid phase enzymolysis product, water and protease to ultrasonic enzymolysis, and then stirring for enzymolysis;
preferably, the temperature of the second enzymolysis is 40-60 ℃, and the total time length is 2-48 hours;
preferably, the protease is used in an amount of not more than 20 parts by weight relative to 100 parts by weight of the solid phase enzymatic hydrolysate on a dry basis.
9. The method according to any one of claims 1 to 8, wherein in step 3, the drying is freeze drying, spray drying or vacuum heat drying.
10. A corn polypeptide meal produced by the method of any one of claims 1-9.
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