CN110547458A - Preparation method of enzymolysis pearl powder - Google Patents
Preparation method of enzymolysis pearl powder Download PDFInfo
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- CN110547458A CN110547458A CN201910870847.8A CN201910870847A CN110547458A CN 110547458 A CN110547458 A CN 110547458A CN 201910870847 A CN201910870847 A CN 201910870847A CN 110547458 A CN110547458 A CN 110547458A
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- 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
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/618—Molluscs, e.g. fresh-water molluscs, oysters, clams, squids, octopus, cuttlefish, snails or slugs
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- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
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- 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
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- C12R2001/00—Microorganisms ; Processes using microorganisms
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- C12R2001/07—Bacillus
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Abstract
the invention discloses a preparation method of enzymolysis pearl powder, belonging to the field of pearl processing. The method of the invention comprises the following steps: the active ingredients in the obtained enzymolysis pearl powder have small molecular weight and are easy to be absorbed by human bodies; the hydrolysis rate of the pearl protein is more than 45 percent, and is improved by more than 10 percent compared with the hydrolysis rate of the conventional single enzyme. The enzymolysis pearl powder has high water solubility, high absorption and utilization rate, high taste quality, simple preparation method and mild process conditions, and is suitable for popularization and application.
Description
Technical Field
The invention belongs to the field of pearl processing, and particularly relates to a preparation method of enzymolysis pearl powder.
Background
the pearl is an ancient organic gem, is mainly produced in mollusks of pearl shellfish and nacre shellfish, and is a mineral (aragonite) bead containing calcium carbonate generated by endocrine action. The pearl contains 17 amino acids such as aspartic acid, threonine, serine, glutamic acid, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, histidine, arginine, proline and the like, and also contains more than 30 trace elements, taurine, abundant vitamins and peptides, has no toxic or side effect, is always used as a treasure for beauty, health care and medicine for thousands of years, however, the pearl is difficult to dissolve in water, is internally eaten and externally used, can only absorb a very small amount of parts of a human body generally, and has no full play of the value.
The development towards the sea economy is an economic development strategy being advocated by the nation. The northern sea is the hometown of south pearl, and pearl resources are very rich; the 'west pearl' is inferior to the 'east pearl' and the 'east pearl' is inferior to the 'south pearl', which is the seawater pearl produced in the north sea. The deep processing form of pearl development at the present stage mainly comprises the following steps: (1) processing pearl into pearl ornament; (2) processing Margarita into Margarita powder and applying to other fields; (3) the pearl active substance is extracted and purified from the pearl by using the modern biotechnology and is utilized. At present, the method for extracting bioactive substances from pearls at home and abroad mainly comprises the following steps: acidolysis method, enzymolysis method, fermentation method and physical hydrolysis method, wherein the acidolysis method and the enzymolysis method are mainly adopted in industrial production. According to the domestic and foreign literature reports, the hydrolysis rate of the acidolysis method reaches 90%, but the equipment requirement is high, the acid liquor is difficult to recover, the environment is polluted, and a large amount of amino acids such as histidine, taurine and the like are easy to damage; the enzymolysis method has the advantages of complete retention of amino acids, especially essential amino acids for human bodies, no environmental pollution, no acid liquid and other impurity residues, small external irritation, safe and reliable use, low hydrolysis rate of pearl protein, large molecular weight of most active ingredients in the enzymolysis of pearls and difficult absorption by human bodies.
Disclosure of Invention
The invention aims to provide a preparation method of enzymolysis pearl powder, which comprises the steps of pretreatment, primary enzymolysis, secondary enzymolysis, fermentation, centrifugation, filtration, post-treatment and the like, wherein active ingredients in the obtained enzymolysis pearl powder have small molecular weight and are easy to be absorbed by human bodies.
the invention is realized by the following technical scheme:
A method for preparing enzymolysis Margarita powder comprises the following steps,
(1) Pretreatment: cleaning and drying pearls, placing the pearls into a crusher for coarse crushing, then placing the crushed pearls into a universal crusher for crushing, sieving the crushed pearls through a 60-80-mesh sieve to obtain coarse pearl powder, and then carrying out micro crushing on the coarse pearl powder by adopting an airflow crusher to obtain 300-mesh and 500-mesh pearl powder; then, the pearl powder is put into 1-5% lactic acid water solution to be soaked for 1-3h to obtain pearl powder solution;
(2) Primary enzymolysis: adjusting pH of the Margarita powder solution to 4.5-5.5, adding 0.05-0.1 wt% of protease A into the Margarita powder solution for enzymolysis at 50-55 deg.C for 3-6 hr, and centrifuging to obtain supernatant I and precipitate I; the protease A is a mixture of papain, trypsin and debitterizing protease according to a mass ratio of 3:1: 2-4;
(3) Secondary enzymolysis: adding water into the precipitate I, stirring and mixing to obtain emulsion, adjusting the pH to 4.5-5.0, adding protease B which is 0.05-0.08% of the emulsion by weight for secondary enzymolysis, wherein the enzymolysis temperature is 45-50 ℃, the enzymolysis time is 2-4h, and centrifuging to obtain a supernatant II and a precipitate II; the protease B is a mixture of papain, trypsin and bromelain according to a mass ratio of 1:3: 2-4;
(4) Fermentation: mixing the supernatant I and the supernatant II to obtain a mixed solution I, adding a mixed bacterial solution of lactobacillus and bacillus with the volume of 5-8% of that of the mixed solution I into the mixed solution I, uniformly mixing, heating to 40-60 ℃, continuously stirring, and carrying out closed fermentation for 48-96h to obtain a fermentation liquid;
(5) Centrifuging: heating the fermentation liquor to 80-85 ℃, keeping the temperature for 5-10min, and then centrifuging by using a centrifugal machine to obtain supernatant fluid to obtain mixed liquor II;
(6) And (3) filtering: filtering the mixed solution II by using a nanofiltration membrane with the aperture of 1-3nm to obtain filtrate;
(7) and (3) post-treatment: and (3) carrying out spray drying on the filtrate by using a spray dryer to obtain the enzymatic hydrolysis pearl powder.
preferably, the pH of the aqueous lactic acid solution is first adjusted to 7.0-8.0 with a sodium bicarbonate solution.
Preferably, the protease A is a mixture of papain, trypsin and debitterizing protease in a mass ratio of 3:1: 3.
Preferably, the protease B is a mixture of papain, trypsin and bromelain in a mass ratio of 1:3: 3.
Preferably, citric acid is used to adjust the pH of the pearl powder solution and emulsion.
As a preferable embodiment, in the step (3), the amount of water added is 2 to 4 times the weight of the precipitate I.
Preferably, the purity of the lactic acid bacteria in the mixed bacterial liquid is 60-70%, and the purity of the bacillus is 30-40%.
preferably, the rotation speed of the centrifugation is 4500-.
Compared with the prior art, the invention has the advantages and beneficial effects that:
(1) The invention adopts lactic acid aqueous solution to treat the pearl powder after being crushed, so that mineral substances in the pearl powder are converted into corresponding lactate which is easily absorbed by human bodies, and therefore, the enzymolysis pearl powder has good absorption effect on the human bodies.
(2) According to the invention, through secondary enzymolysis, and new combined enzyme is adopted for each enzymolysis, the dosage of the enzyme can be reduced, the enzymolysis rate is improved, and the bitter and fishy smell of the product can be reduced; according to the invention, the protease A consisting of papain, trypsin and debitterized protease is adopted for the primary enzymolysis, the protease B consisting of papain, trypsin and bromelain is adopted for the secondary enzymolysis, the dosage of the protease is half of that of a single enzyme, the hydrolysis rate of pearl protein is more than 45%, and is improved by more than 10% compared with that of a conventional single enzyme, so that the combined enzymes of the invention have a synergistic effect, and a good enzymolysis effect is obtained.
(3) the invention also uses lactobacillus and bacillus to ferment after enzymolysis, which can promote the decomposition of protein and monosaccharide into micromolecular active substance, so that the micromolecular active substance in the fermentation liquor accounts for more than 90 percent, amino acid, especially essential amino acid of human body, is completely reserved, and the fermentation effect is good.
(4) The mixed solution is filtered by a nanofiltration membrane with the aperture of 1-3nm, so that macromolecular substances which are not easy to be absorbed by human bodies are intercepted, the proportion of micromolecular active substances in the product is ensured to be more than 95%, and the bitter taste and the fishy smell of the product can be further removed.
(5) the enzymolysis pearl powder has high water solubility, high absorption and utilization rate, high taste quality, simple preparation method and mild process conditions, and is suitable for popularization and application.
Detailed Description
The present invention will be described in further detail with reference to specific examples, but the present invention is not limited to the scope of the present invention.
Example 1
A method for preparing enzymolysis Margarita powder comprises the following steps,
(1) Pretreatment: cleaning Margarita, air drying, coarse crushing in a crusher, pulverizing in a universal pulverizer, sieving with 70 mesh sieve to obtain coarse powder, and micro pulverizing with jet mill to obtain 400 mesh Margarita powder; then, the pearl powder is put into lactic acid water solution with the concentration of 2.5 percent to be soaked for 2.5 hours, and pearl powder solution is obtained. The pH of the aqueous lactic acid solution used was adjusted beforehand to 7.0 with sodium bicarbonate solution.
(2) Primary enzymolysis: adjusting pH of the Margarita powder solution to 5.0 with citric acid, adding 0.07 wt% of protease A into the Margarita powder solution, performing enzymolysis at 52 deg.C for 4 hr, and centrifuging to obtain supernatant I and precipitate I. The protease A is a mixture of papain, trypsin and debitterizing protease in a mass ratio of 3:1: 3.
(3) Secondary enzymolysis: adding water which is 3 times of the weight of the precipitate I into the precipitate I, stirring and mixing to obtain emulsion, adjusting the pH to 4.5 by using citric acid, adding protease B which is 0.07 percent of the weight of the emulsion to carry out secondary enzymolysis, wherein the enzymolysis temperature is 47 ℃, the enzymolysis time is 3 hours, and centrifuging to obtain supernatant II and precipitate II. The protease B is a mixture of papain, trypsin and bromelain according to a mass ratio of 1:3: 3.
(4) fermentation: mixing the supernatant I and the supernatant II to obtain a mixed solution I, adding a mixed bacterial solution of lactobacillus and bacillus with the volume of 6% of that of the mixed solution I into the mixed solution I, uniformly mixing, heating to 50 ℃, continuously stirring, and fermenting in a closed manner for 60 hours to obtain a fermentation liquid. The purity of lactic acid bacteria in the mixed bacterial liquid is 65%, and the purity of bacillus is 35%.
(5) Centrifuging: heating the fermentation liquor to 82 ℃, keeping the temperature for 7min, then centrifuging by using a centrifugal machine, wherein the rotating speed of the centrifugation is 4800r/min, the time is 11min, and taking supernatant fluid to obtain a mixed liquor II.
(6) And (3) filtering: and filtering the mixed solution II by using a nanofiltration membrane with the aperture of 2nm to obtain filtrate.
(7) And (3) post-treatment: and (3) carrying out spray drying on the filtrate by using a spray dryer to obtain the enzymatic hydrolysis pearl powder.
the proportion of micromolecular active substances of the enzymolysis pearl powder prepared by the embodiment is 98%, and the hydrolysis rate of pearl protein is 57%.
Example 2
A method for preparing enzymolysis Margarita powder comprises the following steps,
(1) Pretreatment: cleaning Margarita, air drying, coarse crushing in a crusher, pulverizing in a universal pulverizer, sieving with 65 mesh sieve to obtain coarse Margarita powder, and micro pulverizing with airflow pulverizer to obtain 350 mesh Margarita powder; then, the pearl powder is put into lactic acid water solution with the concentration of 3.5 percent to be soaked for 1.5h, and pearl powder solution is obtained. The pH of the aqueous lactic acid solution used was adjusted beforehand to 7.3 with sodium bicarbonate solution.
(2) Primary enzymolysis: adjusting pH of the Margarita solution to 4.8 with citric acid, adding 0.08 wt% of protease A into the Margarita solution, performing enzymolysis at 53 deg.C for 5 hr, and centrifuging to obtain supernatant I and precipitate I. The protease A is a mixture of papain, trypsin and debitterizing protease in a mass ratio of 3:1: 3.
(3) Secondary enzymolysis: adding water 2.5 times the weight of the precipitate I into the precipitate I, stirring and mixing to obtain emulsion, adjusting the pH to 4.6 by using citric acid, adding protease B0.06% by weight of the emulsion to carry out secondary enzymolysis at 48 ℃ for 3.5h, and centrifuging to obtain supernatant II and precipitate II. The protease B is a mixture of papain, trypsin and bromelain according to a mass ratio of 1:3: 2-4.
(4) Fermentation: and mixing the supernatant I and the supernatant II to obtain a mixed solution I, adding a mixed bacterial solution of lactobacillus and bacillus with the volume of 7% of that of the mixed solution I into the mixed solution I, uniformly mixing, heating to 45 ℃, continuously stirring, and carrying out closed fermentation for 72 hours to obtain a fermentation liquid. The purity of lactic acid bacteria in the mixed bacterial liquid is 67%, and the purity of bacillus is 33%.
(5) Centrifuging: heating the fermentation liquor to 83 ℃, keeping the temperature for 7min, then centrifuging by using a centrifugal machine, wherein the rotation speed of the centrifugation is 4700r/min, the time is 11min, and taking supernatant fluid to obtain mixed liquor II.
(6) And (3) filtering: and filtering the mixed solution II by using a nanofiltration membrane with the aperture of 2nm to obtain filtrate.
(7) and (3) post-treatment: and (3) carrying out spray drying on the filtrate by using a spray dryer to obtain the enzymatic hydrolysis pearl powder.
The proportion of micromolecular active substances of the enzymolysis pearl powder prepared by the embodiment is 98%, and the hydrolysis rate of pearl protein is 55%.
example 3
A method for preparing enzymolysis Margarita powder comprises the following steps,
(1) pretreatment: cleaning Margarita, air drying, coarse crushing in a crusher, pulverizing in a universal pulverizer, sieving with 80 mesh sieve to obtain Margarita coarse powder, and micro pulverizing with jet mill to obtain 500 mesh Margarita powder; then, the pearl powder is put into lactic acid water solution with the concentration of 5 percent to be soaked for 1 hour to obtain pearl powder solution. The pH of the aqueous lactic acid solution used was adjusted beforehand to 7.7 with sodium bicarbonate solution.
(2) Primary enzymolysis: adjusting pH of the Margarita powder solution to 4.5 with citric acid, adding 0.05 wt% protease A into the Margarita powder solution, performing enzymolysis at 50 deg.C for 6 hr, and centrifuging to obtain supernatant I and precipitate I. The protease A is a mixture of papain, trypsin and debitterizing protease in a mass ratio of 3:1: 2.
(3) Secondary enzymolysis: adding water 2 times the weight of the precipitate I into the precipitate I, stirring and mixing to obtain emulsion, adjusting the pH to 4.8 by using citric acid, adding protease B0.05% by weight of the emulsion to carry out secondary enzymolysis at 45 ℃ for 4 hours, and centrifuging to obtain supernatant II and precipitate II. The protease B is a mixture of papain, trypsin and bromelain according to a mass ratio of 1:3: 2-4.
(4) fermentation: and mixing the supernatant I and the supernatant II to obtain a mixed solution I, adding a mixed bacterial solution of lactobacillus and bacillus with the volume of 5% of that of the mixed solution I into the mixed solution I, uniformly mixing, heating to 40 ℃, continuously stirring, and carrying out closed fermentation for 96 hours to obtain a fermentation liquid. The purity of lactic acid bacteria in the mixed bacterial liquid is 60%, and the purity of bacillus is 40%.
(5) Centrifuging: heating the fermentation liquid to 80 ℃, keeping the temperature for 10min, then centrifuging by using a centrifuge at the rotating speed of 4500r/min for 12min, and taking supernatant to obtain a mixed liquid II.
(6) And (3) filtering: and filtering the mixed solution II by using a nanofiltration membrane with the aperture of 1nm to obtain filtrate.
(7) And (3) post-treatment: and (3) carrying out spray drying on the filtrate by using a spray dryer to obtain the enzymatic hydrolysis pearl powder.
The micromolecule active substance ratio of the enzymolysis pearl powder prepared by the embodiment is 97%, and the hydrolysis rate of pearl protein is 53%.
Example 4
A method for preparing enzymolysis Margarita powder comprises the following steps,
(1) Pretreatment: cleaning Margarita, air drying, coarse crushing in a crusher, pulverizing in a universal pulverizer, sieving with 60 mesh sieve to obtain Margarita coarse powder, and micro pulverizing with jet mill to obtain 300 mesh Margarita powder; then, the pearl powder is put into 1 percent lactic acid water solution to be soaked for 3 hours to obtain pearl powder solution. The pH of the aqueous lactic acid solution used was adjusted beforehand to 8.0 with sodium bicarbonate solution.
(2) Primary enzymolysis: adjusting pH of the Margarita powder solution to 5.5 with citric acid, adding 0.1 wt% protease A into the Margarita powder solution, performing enzymolysis at 55 deg.C for 3 hr, and centrifuging to obtain supernatant I and precipitate I. The protease A is a mixture of papain, trypsin and debitterizing protease in a mass ratio of 3:1: 4.
(3) Secondary enzymolysis: adding water 4 times the weight of the precipitate I into the precipitate I, stirring and mixing to obtain emulsion, adjusting the pH to 5.0 by using citric acid, adding protease B0.08% by weight of the emulsion to carry out secondary enzymolysis at 50 ℃ for 2 hours, and centrifuging to obtain supernatant II and precipitate II. The protease B is a mixture of papain, trypsin and bromelain according to a mass ratio of 1:3: 2-4.
(4) Fermentation: and mixing the supernatant I and the supernatant II to obtain a mixed solution I, adding 8% of mixed solution I in volume of mixed solution I into the mixed solution I, uniformly mixing, heating to 60 ℃, continuously stirring, and fermenting in a closed manner for 48 hours to obtain a fermentation liquid. The purity of lactic acid bacteria in the mixed bacterial liquid is 70%, and the purity of bacillus is 30%.
(5) Centrifuging: heating the fermentation liquor to 85 ℃, keeping the temperature for 5min, centrifuging by using a centrifuge at the rotating speed of 5000r/min for 10min, and taking supernatant to obtain a mixed liquor II.
(6) And (3) filtering: and filtering the mixed solution II by using a nanofiltration membrane with the aperture of 3nm to obtain filtrate.
(7) And (3) post-treatment: and (3) carrying out spray drying on the filtrate by using a spray dryer to obtain the enzymatic hydrolysis pearl powder.
The proportion of micromolecular active substances of the enzymolysis pearl powder prepared by the embodiment is 96 percent, and the hydrolysis rate of pearl protein is more than 48 percent.
Claims (8)
1. A preparation method of enzymolysis pearl powder is characterized by comprising the following steps,
(1) Pretreatment: cleaning and drying pearls, placing the pearls into a crusher for coarse crushing, then placing the crushed pearls into a universal crusher for crushing, sieving the crushed pearls through a 60-80-mesh sieve to obtain coarse pearl powder, and then carrying out micro crushing on the coarse pearl powder by adopting an airflow crusher to obtain 300-mesh and 500-mesh pearl powder; then, the pearl powder is put into 1-5% lactic acid water solution to be soaked for 1-3h to obtain pearl powder solution;
(2) primary enzymolysis: adjusting pH of the Margarita powder solution to 4.5-5.5, adding 0.05-0.1 wt% of protease A into the Margarita powder solution for enzymolysis at 50-55 deg.C for 3-6 hr, and centrifuging to obtain supernatant I and precipitate I; the protease A is a mixture of papain, trypsin and debitterizing protease according to a mass ratio of 3:1: 2-4;
(3) secondary enzymolysis: adding water into the precipitate I, stirring and mixing to obtain emulsion, adjusting the pH to 4.5-5.0, adding protease B which is 0.05-0.08% of the emulsion by weight for secondary enzymolysis, wherein the enzymolysis temperature is 45-50 ℃, the enzymolysis time is 2-4h, and centrifuging to obtain a supernatant II and a precipitate II; the protease B is a mixture of papain, trypsin and bromelain according to a mass ratio of 1:3: 2-4;
(4) fermentation: mixing the supernatant I and the supernatant II to obtain a mixed solution I, adding a mixed bacterial solution of lactobacillus and bacillus with the volume of 5-8% of that of the mixed solution I into the mixed solution I, uniformly mixing, heating to 40-60 ℃, continuously stirring, and carrying out closed fermentation for 48-96h to obtain a fermentation liquid;
(5) Centrifuging: heating the fermentation liquor to 80-85 ℃, keeping the temperature for 5-10min, and then centrifuging by using a centrifugal machine to obtain supernatant fluid to obtain mixed liquor II;
(6) And (3) filtering: filtering the mixed solution II by using a nanofiltration membrane with the aperture of 1-3nm to obtain filtrate;
(7) and (3) post-treatment: and (3) carrying out spray drying on the filtrate by using a spray dryer to obtain the enzymatic hydrolysis pearl powder.
2. The method for preparing enzymatically hydrolyzed pearl powder according to claim 1, wherein the pH of the aqueous lactic acid solution is first adjusted to 7.0 to 8.0 with a sodium bicarbonate solution.
3. the method for preparing the enzymatic pearl powder according to claim 1, wherein the protease A is a mixture of papain, trypsin and debitterizing protease in a mass ratio of 3:1: 3.
4. the method for preparing the enzymatic pearl powder according to claim 3, wherein the protease B is a mixture of papain, trypsin and bromelain in a mass ratio of 1:3: 3.
5. The process for the preparation of enzymatically hydrolyzed pearl powder, according to claim 1, wherein the pH of the pearl powder solution and emulsion is adjusted with citric acid.
6. The method for preparing enzymatically hydrolyzed pearl powder according to claim 1, wherein, in the step (3), the amount of water added is 2 to 4 times the weight of the precipitate i.
7. The method for preparing the enzymatic pearl powder according to claim 1, wherein the purity of the lactic acid bacteria in the mixed bacterial liquid is 60-70%, and the purity of the bacillus is 30-40%.
8. The method for preparing enzymatic pearl powder according to claim 1, wherein the centrifugation speed is 4500-.
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