CN116694719A - Preparation method and application of small-molecule corn oligopeptide - Google Patents
Preparation method and application of small-molecule corn oligopeptide Download PDFInfo
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- 102000035195 Peptidases Human genes 0.000 abstract description 11
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 abstract description 8
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 abstract description 8
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 abstract description 7
- 235000004279 alanine Nutrition 0.000 abstract description 7
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
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- Proteomics, Peptides & Aminoacids (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
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Abstract
The invention relates to a preparation method and application of a small-molecule corn oligopeptide. The preparation method comprises the steps of heating pretreatment of corn gluten meal, stepwise multi-stage protease enzymolysis, centrifugal deslagging, filter membrane filtration, active carbon decolorization, ultrafiltration concentration, spray drying and the like. The multi-stage protease enzymolysis step by step adopts a plurality of proteases, and the corn oligopeptide with unique amino acid and polypeptide composition is obtained through the combined enzymolysis of compound proteases. The obtained corn oligopeptide has the advantages of remarkably improved contents of small molecular weight substances and alanine and leucine, good taste and easy acceptance, and is more favorable for human body absorption and utilization.
Description
Technical Field
The invention relates to a method for preparing small-molecule corn oligopeptide by adopting composite protease enzymolysis and application thereof, belonging to the technical field of corn peptide production.
Background
Corn is a main grain crop in China, and is rich in nutrients such as starch, protein, vitamins and the like. Corn gluten meal, also known as corn gluten meal, is a by-product produced in the process of producing corn starch. Because the natural zein has very large molecular weight and poor water solubility, the nutrient substances are difficult to be effectively absorbed and utilized, and the nutrition value is not fully reflected. Research shows that the small molecular peptide obtained after enzymolysis of corn protein has the functions of dispelling the effects of alcohol, protecting liver, resisting oxidation, delaying aging, improving immunity and the like, wherein the small molecular peptide with the molecular mass of less than 1000Da is easier to be absorbed and utilized by human bodies to exert functional activity.
The corn protein has unique amino acid composition different from other grain crops, and the corn oligopeptide is rich in various amino acids such as leucine and alanine and small peptide substances, and the alanine and the leucine can play an important role in alleviating hangover and protecting liver by enhancing the activities of alcohol dehydrogenase and acetaldehyde dehydrogenase in vivo, promoting alcohol metabolism, relieving adverse reaction and other symptoms caused by drinking. However, the existing preparation process of the corn oligopeptide has the problems of low yield of the low molecular weight oligopeptide, bitter taste and the like.
Disclosure of Invention
The main purpose of the invention is to provide the corn oligopeptide with high yield of small molecular oligopeptide, improved yield of alanine and leucine and good taste.
The invention is realized by the following technical scheme:
the preparation method of the corn oligopeptide is characterized by comprising the steps of corn yellow powder heating pretreatment, stepwise multi-stage protease enzymolysis, centrifugal deslagging, filter membrane filtration, active carbon decolorization, ultrafiltration concentration and spray drying.
Specifically, the heating pretreatment step includes: weighing a certain weight of corn gluten, adding the corn gluten into a container, adding deionized water according to the weight ratio of the corn gluten to the deionized water of 1:8-1:12, fully stirring and mixing at room temperature, heating to 80-95 ℃, preserving the heat for 20-50 minutes in a stirring state, centrifuging at 2500-4000rpm for 10-15 minutes, discarding the supernatant, and transferring the precipitate into an enzymolysis tank.
Preferably, the stepwise multi-stage protease enzymolysis comprises any one, two or three of acid protease, alkaline protease, neutral protease, papain and flavourzyme.
Preferably, the stepwise multistage protease enzymolysis is performed according to the following steps: 1) Enzymolysis with acid protease; 2) After the enzymolysis of the acid protease is finished, centrifuging to obtain a supernatant, and adding neutral protease, papain and/or flavourzyme into the supernatant to carry out continuous enzymolysis; 3) After the enzymolysis is finished, centrifuging to obtain supernatant, and adding alkaline protease into the supernatant to continue the enzymolysis.
Preferably, the detailed steps of the stepwise multistage protease enzymolysis are as follows: (1) Adjusting pH to 2-4.2 with dilute hydrochloric acid solution, adding acid protease according to the weight ratio of 0.8-1.5% of corn gluten meal, and performing enzymolysis for 2-4 hours at 35-45deg.C under stirring at 100 rpm. And (3) boiling the enzymolysis liquid for 7-10 minutes after the enzymolysis is finished, cooling to room temperature, and centrifugally separating the enzymolysis liquid. (2) Regulating pH of supernatant to 6.2-7.2 with NaOH 1 wt%, adding neutral protease, papain and flavourzyme into supernatant, and calculating the adding ratio of the three enzymes according to the weight of corn gluten meal 0.3-0.5 wt% respectively. And (3) carrying out enzymolysis for 2-4 hours at 40-60 ℃ under the stirring condition of 100 rpm. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, cooling to room temperature, and centrifugally separating the enzymolysis liquid. (3) The pH of the supernatant is adjusted to 10.0-11.0 by using 1%wt NaOH, and alkaline protease is added into the supernatant according to the proportion of 0.8-1.5%wt of the mass of the maize yellow powder. And (3) carrying out enzymolysis for 2-4 hours at 55-65 ℃ under the stirring condition of 100 rpm. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, and cooling to room temperature.
Preferably, the addition amount of the acid protease and the alkaline protease is 1 percent by weight of the mass of the corn gluten meal, and the addition amount of the neutral protease, the papain and the flavor protease is 0.3 percent by weight of the mass of the corn gluten meal.
Preferably, the enzymolysis time is 4 hours.
The invention also provides the small-molecule corn oligopeptide prepared by the method.
Further, the invention also provides a food, health care product or composition containing the small molecule corn oligopeptide.
Furthermore, the invention also provides the application of any one of the methods, the small-molecule corn oligopeptide and the food, the health-care product or the composition containing the small-molecule corn oligopeptide in preparation of the anti-alcohol and liver-protecting product.
The invention has the beneficial effects that:
in the invention, a plurality of proteases are adopted in the enzymolysis process of the corn oligopeptide, and the corn oligopeptide with unique amino acid and polypeptide composition is obtained through the combined enzymolysis of compound proteases. The detection result shows that the small molecular weight substances in the obtained corn oligopeptide are obviously improved, wherein the small molecular weight of less than 1000 daltons is more than 95%, and the individual scheme is 97%, so that the corn oligopeptide is more beneficial to human body absorption and utilization. In addition, the content of alanine and leucine in the prepared corn oligopeptide is also obviously improved, and the obtained corn oligopeptide has good taste and insignificant bitter taste, so that the audience range of the corn oligopeptide is greatly increased, and the corn oligopeptide is more easily accepted by people.
Detailed Description
The invention is further described below with reference to examples.
Example 1
(1) 1kg of corn gluten meal is weighed and added into a container, 10kg of deionized water is added, and the mixture is stirred and mixed uniformly at room temperature. After thorough mixing, the liquid containing the corn gluten meal was heated to 90 ℃ and incubated for 40 minutes with stirring, centrifuged at 3500rpm for 10 minutes, the supernatant discarded, and the precipitate collected and transferred to an enzymatic tank.
(2) The pH value is regulated to 4.1 by adopting a dilute hydrochloric acid solution, and acid protease is added according to the proportion of 1 wt% of the mass of the maize yellow powder, and the enzymolysis is carried out for 4 hours under the stirring condition of 100rpm at 40 ℃. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, cooling to room temperature, and centrifugally separating the enzymolysis liquid.
(3) The pH of the supernatant is regulated to 6.9 by adopting 1%wt NaOH, neutral protease, papain and flavourzyme are added into the supernatant, and the adding proportion of the three enzymes is calculated according to 0.3%wt of the mass of the maize yellow powder respectively. And (3) carrying out enzymolysis for 3.5 hours at 50 ℃ under the stirring condition of 100 rpm. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, cooling to room temperature, and centrifugally separating the enzymolysis liquid.
(4) The pH of the supernatant was adjusted to 10.5 with 1% by weight NaOH, and alkaline protease was added to the supernatant in a proportion of 1% by weight based on the mass of the corn meal. And (3) carrying out enzymolysis for 4 hours at 60 ℃ under the stirring condition of 100 rpm. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, and cooling to room temperature.
(5) Centrifuging at 5000rpm for 20min, and removing residues. The supernatant was filtered through a filter membrane and transferred to a new vessel, and the pH of the supernatant was adjusted to 7.0 with dilute hydrochloric acid.
(6) Activated carbon is added according to the proportion of 2% (w/v) of the volume of the supernatant, and the activated carbon is adopted for filtering and decoloring for about 1 hour.
(7) And (3) carrying out ultrafiltration concentration and spray drying on the decolorized solution to obtain the micromolecular corn oligopeptide powder.
Example 2
(1) 1kg of corn gluten meal is weighed and added into a container, 10kg of deionized water is added, and the mixture is stirred and mixed uniformly at room temperature. After thorough mixing, the liquid containing the corn gluten meal was heated to 80 ℃ and incubated for 40 minutes with stirring, centrifuged at 3500rpm for 10 minutes, the supernatant discarded, and the precipitate collected and transferred to an enzymatic tank.
(2) The pH value is regulated to 3.5 by adopting a dilute hydrochloric acid solution, and acid protease is added according to the proportion of 1.2 wt% of the mass of the maize yellow powder, and the enzymolysis is carried out for 4 hours under the stirring condition of 100rpm at 40 ℃. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, cooling to room temperature, and centrifugally separating the enzymolysis liquid.
(3) The pH of the supernatant is regulated to 7.1 by adopting 1%wt NaOH, neutral protease, papain and flavourzyme are added into the supernatant, and the adding proportion of the three enzymes is calculated according to 0.4%wt of the mass of the maize yellow powder respectively. And (3) carrying out enzymolysis for 3.5 hours at 50 ℃ under the stirring condition of 100 rpm. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, cooling to room temperature, and centrifugally separating the enzymolysis liquid.
(4) The pH of the supernatant was adjusted to 10.0 with 1% wt NaOH and alkaline protease was added to the supernatant in a proportion of 1.2% wt based on the mass of the corn meal. And (3) carrying out enzymolysis for 4 hours at 60 ℃ under the stirring condition of 100 rpm. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, and cooling to room temperature.
(5) Centrifuging at 5000rpm for 20min, and removing residues. The supernatant was filtered through a filter membrane and transferred to a new vessel, and the pH of the supernatant was adjusted to 7.0 with dilute hydrochloric acid.
(6) Activated carbon is added according to the proportion of 2% (w/v) of the volume of the supernatant, and the activated carbon is adopted for filtering and decoloring for about 1 hour.
(7) And (3) carrying out ultrafiltration concentration and spray drying on the decolorized solution to obtain the micromolecular corn oligopeptide powder.
Example 3
(1) 1kg of corn gluten meal is weighed and added into a container, 10kg of deionized water is added, and the mixture is stirred and mixed uniformly at room temperature. After thorough mixing, the liquid containing the corn gluten meal was heated to 90 ℃ and incubated for 40 minutes with stirring, centrifuged at 3500rpm for 10 minutes, the supernatant discarded, and the precipitate collected and transferred to an enzymatic tank.
(2) The pH value is regulated to 4.1 by adopting a dilute hydrochloric acid solution, and acid protease is added according to the proportion of 1 wt% of the mass of the maize yellow powder, and the enzymolysis is carried out for 3 hours under the stirring condition of 100rpm at 40 ℃. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, cooling to room temperature, and centrifugally separating the enzymolysis liquid.
(3) The pH of the supernatant is regulated to 6.9 by adopting 1%wt NaOH, neutral protease, papain and flavourzyme are added into the supernatant, and the adding proportion of the three enzymes is calculated according to 0.3%wt of the mass of the maize yellow powder respectively. And (3) carrying out enzymolysis for 3 hours at 50 ℃ under the stirring condition of 100 rpm. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, cooling to room temperature, and centrifugally separating the enzymolysis liquid.
(4) The pH of the supernatant was adjusted to 10.5 with 1% by weight NaOH, and alkaline protease was added to the supernatant in a proportion of 1% by weight based on the mass of the corn meal. And (3) carrying out enzymolysis for 3 hours at 60 ℃ under the stirring condition of 100 rpm. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, and cooling to room temperature.
(5) Centrifuging at 5000rpm for 20min, and removing residues. The supernatant was filtered through a filter membrane and transferred to a new vessel, and the pH of the supernatant was adjusted to 7.0 with dilute hydrochloric acid.
(6) Activated carbon is added according to the proportion of 2% (w/v) of the volume of the supernatant, and the activated carbon is adopted for filtering and decoloring for about 1 hour.
(7) And (3) carrying out ultrafiltration concentration and spray drying on the decolorized solution to obtain the micromolecular corn oligopeptide powder.
Example 4
(1) 1kg of corn gluten meal is weighed and added into a container, 10kg of deionized water is added, and the mixture is stirred and mixed uniformly at room temperature. After thorough mixing, the liquid containing the corn gluten meal was heated to 90 ℃ and incubated for 40 minutes with stirring, centrifuged at 3500rpm for 10 minutes, the supernatant discarded, and the precipitate collected and transferred to an enzymatic tank.
(2) The pH value is regulated to 4.1 by adopting a dilute hydrochloric acid solution, and acid protease is added according to the proportion of 1 wt% of the mass of the maize yellow powder, and the enzymolysis is carried out for 2 hours under the stirring condition of 100rpm at 40 ℃. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, cooling to room temperature, and centrifugally separating the enzymolysis liquid.
(3) The pH of the supernatant is regulated to 6.9 by adopting 1%wt NaOH, neutral protease, papain and flavourzyme are added into the supernatant, and the adding proportion of the three enzymes is calculated according to 0.3%wt of the mass of the maize yellow powder respectively. And (3) carrying out enzymolysis for 2 hours at 50 ℃ under the stirring condition of 100 rpm. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, cooling to room temperature, and centrifugally separating the enzymolysis liquid.
(4) The pH of the supernatant was adjusted to 10.5 with 1% by weight NaOH, and alkaline protease was added to the supernatant in a proportion of 1% by weight based on the mass of the corn meal. And (3) carrying out enzymolysis for 2 hours at 60 ℃ under the stirring condition of 100 rpm. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, and cooling to room temperature.
(5) Centrifuging at 5000rpm for 20min, and removing residues. The supernatant was filtered through a filter membrane and transferred to a new vessel, and the pH of the supernatant was adjusted to 7.0 with dilute hydrochloric acid.
(6) Activated carbon is added according to the proportion of 2% (w/v) of the volume of the supernatant, and the activated carbon is adopted for filtering and decoloring for about 1 hour.
(7) And (3) carrying out ultrafiltration concentration and spray drying on the decolorized solution to obtain the micromolecular corn oligopeptide powder.
Comparative example 1
(1) 1kg of corn gluten meal is weighed and added into a container, 10kg of deionized water is added, and the mixture is stirred and mixed uniformly at room temperature. After thorough mixing, the liquid containing the corn gluten meal was heated to 90 ℃ and incubated for 40 minutes with stirring, centrifuged at 3500rpm for 10 minutes, the supernatant discarded, and the precipitate collected and transferred to an enzymatic tank.
(2) And (3) regulating the pH to 4.1 by adopting a dilute hydrochloric acid solution, adding bacillus subtilis acid protease according to the proportion of 1% by weight of corn gluten meal, and carrying out enzymolysis for 4 hours at 40 ℃ under the stirring condition of 100 rpm. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, cooling to room temperature, and centrifugally separating the enzymolysis liquid.
(3) The pH of the supernatant is regulated to 6.9 by using 1% wt NaOH, neutral protease is added according to the proportion of 0.9% wt of the mass of the maize yellow powder, and enzymolysis is carried out for 3.5 hours under the stirring condition of 100rpm at 50 ℃. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, cooling to room temperature, and centrifugally separating the enzymolysis liquid.
(4) The pH of the supernatant was adjusted to 10.5 with 1% by weight NaOH, and alkaline protease was added to the supernatant in a proportion of 1% by weight based on the mass of the corn meal. And (3) carrying out enzymolysis for 4 hours at 60 ℃ under the stirring condition of 100 rpm. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, and cooling to room temperature.
(5) Centrifuging at 5000rpm for 20min, and removing residues. The supernatant was filtered through a filter membrane and transferred to a new vessel, and the pH of the supernatant was adjusted to 7.0 with dilute hydrochloric acid.
(6) Activated carbon is added according to the proportion of 2% (w/v) of the volume of the supernatant, and the activated carbon is adopted for filtering and decoloring for about 1 hour.
(7) And (3) carrying out ultrafiltration concentration and spray drying on the decolorized solution to obtain the micromolecular corn oligopeptide powder.
Comparative example 2
(1) 1kg of corn gluten meal is weighed and added into a container, 10kg of deionized water is added, and the mixture is stirred and mixed uniformly at room temperature. After thorough mixing, the liquid containing the corn gluten meal was heated to 90 ℃ and incubated for 40 minutes with stirring, centrifuged at 3500rpm for 10 minutes, the supernatant discarded, and the precipitate collected and transferred to an enzymatic tank.
(2) And (3) regulating the pH to 4.1 by adopting a dilute hydrochloric acid solution, adding bacillus subtilis acid protease according to the proportion of 1% by weight of corn gluten meal, and carrying out enzymolysis for 4 hours at 40 ℃ under the stirring condition of 100 rpm. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, cooling to room temperature, and centrifugally separating the enzymolysis liquid.
(3) The pH of the supernatant is regulated to 6.9 by adopting 1%wt NaOH, neutral protease, papain and flavourzyme are added into the supernatant, and the adding proportion of the three enzymes is calculated according to 0.3%wt of the mass of the maize yellow powder respectively. And (3) carrying out enzymolysis for 3.5 hours at 50 ℃ under the stirring condition of 100 rpm. And (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, cooling to room temperature, and centrifugally separating the enzymolysis liquid.
(4) Centrifuging at 5000rpm for 20min, and removing residues. The supernatant was filtered through a filter membrane and transferred to a new vessel, and the pH of the supernatant was adjusted to 7.0 with dilute hydrochloric acid.
(5) Activated carbon is added according to the proportion of 2% (w/v) of the volume of the supernatant, and the activated carbon is adopted for filtering and decoloring for about 1 hour.
(6) And (3) carrying out ultrafiltration concentration and spray drying on the decolorized solution to obtain the micromolecular corn oligopeptide powder.
Unless otherwise specified, ultrafiltration concentration, spray drying and the like in the present invention are all conventional preparation processes, and have no particular influence on the results of the practice of the present invention.
The molecular weight distribution of the prepared corn oligopeptide powder was detected by referring to GB/T22729-2008, and the amino acid content of the oligopeptide was analyzed by using an L-8900 full-automatic amino acid analyzer, and the detection results are shown in tables 1 and 2, respectively.
TABLE 1 molecular weight distribution of small molecule corn oligopeptides obtained in different examples and comparative examples
Table 2 different examples and comparative examples the glutamic acid and leucine content and sensory evaluation of small molecule corn oligopeptides were obtained
According to the results in tables 1 and 2, the content of small molecular weight substances in the corn oligopeptide can be obviously improved by adopting the stepped multi-stage enzymolysis, and particularly, the enzymolysis effect is optimal by combining three steps of acid protease, neutral protease, papain, flavourzyme and alkaline protease. The alkaline protease enzymolysis step which lacks the third step or the second step adopts neutral protease enzymolysis only, and papain and flavourzyme are not added, so that the content of the small molecular weight oligopeptide is obviously reduced. The optimized three-step enzymolysis scheme and protease combination can obviously improve the content of the small molecular weight oligopeptide with the molecular weight smaller than 1000Da in the enzymolysis liquid, and the prepared corn oligopeptide is easier to be absorbed and utilized by human bodies.
In addition, the alanine and leucine content in the prepared corn oligopeptide is also obviously improved. The prior art has demonstrated that alanine and leucine promote ethanol metabolism by enhancing the activity of alcohol dehydrogenase and acetaldehyde dehydrogenase in vivo. Therefore, the corn oligopeptide prepared by the invention can be used for preparing anti-alcohol and liver-protecting products, and has important application value in the field of health products.
The above embodiments are only for illustrating the present invention and not for limiting the technical solutions described in the present invention, and thus, although the present invention has been described in detail with reference to the above embodiments, it should be understood by those skilled in the art that the present invention may be modified or substituted by equivalent ones without departing from the spirit and scope of the present invention and all modifications thereof are intended to be covered in the scope of the claims of the present invention.
Claims (10)
1. The preparation method of the small molecular corn oligopeptide is characterized by comprising the steps of corn gluten meal heating pretreatment, stepwise multi-stage protease enzymolysis, centrifugal deslagging, filter membrane filtration, active carbon decolorization, ultrafiltration concentration and spray drying, and is specifically characterized in that:
(1) Weighing a certain weight of corn gluten, adding the corn gluten into a container, adding deionized water according to the weight ratio of the corn gluten to the deionized water of 1:8-1:12, fully stirring and mixing at room temperature, heating to 80-95 ℃, preserving heat for 20-50 minutes in a stirring state, centrifuging at 2500-4000rpm for 10-15 minutes, discarding the supernatant, and transferring the precipitate into an enzymolysis tank;
(2) The stepwise multistage protease enzymolysis is carried out according to the following steps: 1) Enzymolysis with acid protease; 2) After the enzymolysis of the acid protease is finished, centrifuging to obtain a supernatant, and adding any one, two or three of neutral protease, papain and flavourzyme into the supernatant for continuous enzymolysis; 3) After the enzymolysis is finished, centrifuging to obtain supernatant, and adding alkaline protease into the supernatant to continue the enzymolysis;
(3) Centrifuging to remove residues after enzymolysis, filtering the supernatant with a filter membrane, and adjusting the pH of the filtrate to 6.8-7.2 with dilute hydrochloric acid;
(4) Adding active carbon according to the proportion of 1.5-3% (w/v) of the volume of the filtrate, and filtering and decoloring by adopting the active carbon;
(5) And (3) carrying out ultrafiltration concentration and spray drying on the decolorized solution to obtain the micromolecular corn oligopeptide powder.
2. The method according to claim 1, characterized in that: the step (2) is specifically carried out by adopting a combination of neutral protease, papain and flavourzyme.
3. The method according to any one of claims 1-2, wherein: the specific steps of the step (2) comprise:
1) Adjusting pH to 2-4.2 with dilute hydrochloric acid solution, adding acid protease according to the weight ratio of 0.8-1.5% of corn gluten meal, and performing enzymolysis for 2-4 hours at 35-45deg.C under stirring at 100 rpm; boiling the enzymolysis liquid for 7-10 minutes after enzymolysis is finished, cooling to room temperature, and centrifugally separating the enzymolysis liquid;
2) Regulating the pH of the supernatant to 6.2-7.2 by adopting 1%wt NaOH, adding neutral protease, papain and flavourzyme into the supernatant, wherein the adding proportion of the neutral protease, papain and flavourzyme is respectively calculated according to 0.3-0.5%wt of the corn gluten meal; enzymolysis is carried out for 2-4 hours under the stirring condition of 100rpm at 40-60 ℃; boiling the enzymolysis liquid for 7 minutes after enzymolysis is finished, cooling to room temperature, and centrifugally separating the enzymolysis liquid;
3) Adjusting the pH of the supernatant to 10.0-11.0 by using 1%wt NaOH, and adding alkaline protease into the supernatant according to the proportion of 0.8-1.5%wt of the corn gluten meal; enzymolysis is carried out for 2 to 4 hours under the stirring condition of 100rpm at 55 to 65 ℃; and (3) boiling the enzymolysis liquid for 7 minutes after the enzymolysis is finished, and cooling to room temperature.
4. The method according to any one of claims 1-4, wherein: the addition amount of the acid protease and the alkaline protease is 1% wt of the mass of the corn gluten meal: the addition amount of the neutral protease, the papain and the flavourzyme is 0.3% wt of the mass of the maize yellow meal respectively.
5. The method according to any one of claims 1-4, wherein: 1) The pH value of the enzyme is 4.1, the enzymolysis temperature is 40 ℃, and the enzymolysis time is 4 hours.
6. The method according to any one of claims 1-4, wherein: 2) The pH value of the enzyme is 6.9, the enzymolysis temperature is 50 ℃, and the enzymolysis time is 3.5 hours.
7. The method according to any one of claims 1-4, wherein: 3) The pH value of the enzyme is 10.5, the enzymolysis temperature is 60 ℃, and the enzymolysis time is 4 hours.
8. A small molecule corn oligopeptide prepared according to the method of any one of claims 1-7.
9. A food product, nutraceutical or composition comprising the small molecule corn oligopeptide of claim 8.
10. Use of the method of any one of claims 1-7, the small molecule corn oligopeptide of claim 8, or the food, health product, composition of claim 9 for the preparation of an anti-hangover and liver-protecting product.
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