CN101869169B - Method for preparing fish oligopeptide from gurry by combining fermentation and membrane technology - Google Patents
Method for preparing fish oligopeptide from gurry by combining fermentation and membrane technology Download PDFInfo
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- 241000251468 Actinopterygii Species 0.000 title claims abstract description 78
- 108010038807 Oligopeptides Proteins 0.000 title claims abstract description 38
- 102000015636 Oligopeptides Human genes 0.000 title claims abstract description 38
- 238000000855 fermentation Methods 0.000 title claims abstract description 21
- 230000004151 fermentation Effects 0.000 title claims abstract description 21
- 238000005516 engineering process Methods 0.000 title claims abstract description 11
- 239000012528 membrane Substances 0.000 title abstract description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 21
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 20
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 20
- 102000008186 Collagen Human genes 0.000 claims abstract description 17
- 108010035532 Collagen Proteins 0.000 claims abstract description 17
- 229920001436 collagen Polymers 0.000 claims abstract description 17
- 229960005188 collagen Drugs 0.000 claims abstract description 17
- 238000001914 filtration Methods 0.000 claims abstract description 9
- 230000001954 sterilising Effects 0.000 claims abstract description 8
- 238000007670 refining Methods 0.000 claims abstract description 7
- 238000009777 vacuum freeze-drying Methods 0.000 claims abstract description 5
- 238000001471 micro-filtration Methods 0.000 claims abstract description 4
- 238000001728 nano-filtration Methods 0.000 claims abstract description 3
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 3
- 239000000463 material Substances 0.000 claims description 23
- 235000021190 leftovers Nutrition 0.000 claims description 13
- 241000194103 Bacillus pumilus Species 0.000 claims description 10
- 239000003292 glue Substances 0.000 claims description 10
- 241000894006 Bacteria Species 0.000 claims description 9
- 239000012141 concentrate Substances 0.000 claims description 9
- 241001507822 Aspergillus terricola Species 0.000 claims description 8
- 238000010612 desalination reaction Methods 0.000 claims description 8
- 239000012530 fluid Substances 0.000 claims description 8
- 239000012535 impurity Substances 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 7
- 241000186046 Actinomyces Species 0.000 claims description 6
- 239000002054 inoculum Substances 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 6
- 238000010828 elution Methods 0.000 claims description 2
- 230000001105 regulatory Effects 0.000 claims description 2
- 239000000047 product Substances 0.000 abstract description 18
- 238000004519 manufacturing process Methods 0.000 abstract description 12
- 230000000694 effects Effects 0.000 abstract description 7
- 238000005265 energy consumption Methods 0.000 abstract description 4
- 238000004042 decolorization Methods 0.000 abstract 1
- 238000011033 desalting Methods 0.000 abstract 1
- 230000000249 desinfective Effects 0.000 abstract 1
- 239000000413 hydrolysate Substances 0.000 abstract 1
- 238000005406 washing Methods 0.000 abstract 1
- 238000000034 method Methods 0.000 description 14
- 102000004190 Enzymes Human genes 0.000 description 7
- 108090000790 Enzymes Proteins 0.000 description 7
- 239000000203 mixture Substances 0.000 description 6
- MVORZMQFXBLMHM-QWRGUYRKSA-N (2S)-6-amino-2-[[(2S)-2-[(2-aminoacetyl)amino]-3-(1H-imidazol-5-yl)propanoyl]amino]hexanoic acid Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CN=CN1 MVORZMQFXBLMHM-QWRGUYRKSA-N 0.000 description 5
- 108010038983 glycyl-histidyl-lysine Proteins 0.000 description 5
- 238000006460 hydrolysis reaction Methods 0.000 description 5
- 238000010792 warming Methods 0.000 description 5
- 229910052799 carbon Inorganic materials 0.000 description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 4
- 238000001035 drying Methods 0.000 description 4
- 239000000706 filtrate Substances 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 239000002699 waste material Substances 0.000 description 4
- 238000003809 water extraction Methods 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 3
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- 241000251730 Chondrichthyes Species 0.000 description 2
- 102000033147 ERVK-25 Human genes 0.000 description 2
- 108091005771 Peptidases Proteins 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 230000003078 antioxidant Effects 0.000 description 2
- 230000001580 bacterial Effects 0.000 description 2
- 230000000975 bioactive Effects 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 238000010298 pulverizing process Methods 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 210000000936 Intestines Anatomy 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000003213 activating Effects 0.000 description 1
- 230000001464 adherent Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000002929 anti-fatigue Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 235000019658 bitter taste Nutrition 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000000875 corresponding Effects 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000003301 hydrolyzing Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000006210 lotion Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000036633 rest Effects 0.000 description 1
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Abstract
The invention relates to a method for preparing fish oligopeptide from gurry by combining fermentation and membrane technology, which comprises the following steps of: (1) crushing the gurry, adding water to make the water account for 40 to 70 weight percent of the gurry, sterilizing for 10 to 30 minutes at the temperature of 115 DEG C, adding mixed strains in an amount which accounts for 2 to 8 percent of dry weight of the gurry to perform fermentation after the temperature is reduced to between 25 and 45 DEG C, adjusting the pH value to between 6.5 and 8.0 and performing fermentation for 36 to 96 hours; (2) after extracting collagen oligopeptide or mixed protein oligopeptide from fermentation liquor by washing, performing decolorization and rough filtration, and then purifying, disinfecting, refining, concentrating and desalting hydrolysate by using a microfiltration film, an ultrafiltration film and a nanofiltration film; and (3) further concentrating liquor concentrated by the third-level film by circulating at a low temperature, and then performing vacuum freeze drying to obtain fish collagen oligopeptide or fish mixed protein oligopeptide. The method effectively solves the problems of high cost, high energy consumption, and low purity and activity of products existing in the conventional fish oligopeptide production process.
Description
Technical field
The present invention relates to a kind of method of producing fish oligopeptide, particularly relate to the mixed culture fermentation of a kind of employing mixed bacteria, take fish offal material such as fish head, fish scale, fish-skin, fish tail, shark's fin adherent meats as raw material, the binding film separation process technique, the method for producing Isin glue collagen oligopeptide or fish mixed protein oligopeptide at a production line.
Background technology
China is world fish culture big country, accounts for about 1/3 of Gross World Product, occupies the first in the world.Although possess advantageous aquatic products fish resource advantage, the utilization of fish resource still rests on the comparatively original stage, substantially with the need of fresh and alive form sale for food and drink.Though small part is arranged through being processed into the products such as fish gruel, can and Cheng Ganyu, all belongs to elementary processing mode.In fish preliminary working process, produce a large amount of preliminary working leftover bits and pieces, account for 30%～40% of fish gross weight such as fish head, fish tail, fish scale, fish-skin, fish guts etc., these leftover bits and pieces or directly abandon, simply being processed as feed sells at a low price, not only value is not fully utilized, even also pollutes the environment.And the flesh of fish that fish head, fish tail, fish intestines and shark's fin adhere to is rich in multiple proteins, fish scale and fish-skin contain abundant Isin glue collagen, if fully development and use can be turned waste into wealth, prepare respectively fish mixed protein oligopeptide and the collagen oligopeptide with high added value.
Oligopeptide is a kind of important bioactivator, be easily absorbed by the body and have higher physiologically active, be mainly manifested in: regain one's strength of body fast, regulate the aspects such as immunity, anti-hypertension, anti-oxidant, antifatigue, can be widely used in the industry fields such as food, health products, cosmetics, and has widely consumption market, abroad and form huge consumption market, domestic demand is also enlarging gradually.
Chinese patent: CN 200810195646.4 " endogenous enzymes prepares the technical study of bioactive micro peptide " discloses a kind of technique of utilizing endogenous enzymes hydrolysis marine products Optimization of Low Value Fish waste material to prepare bioactive micro peptide, this technique comprises the endogenous enzymes preliminary treatment, enzymolysis, screen filtration, concentrated, drying and other steps, it is low that this scheme exists endogenous enzymes hydrolysis power, the endogenous enzymes of fish body itself is not enough to be hydrolyzed corresponding fish offal, lack and separate, removal of impurities and subtractive process, the long peptide chain and the little molecule free amino acid that fully are hydrolyzed that exist in the enzymolysis liquid are not effectively separated and removal of impurities, the purity of biologically active peptide is not high, the problems such as concentrated and dry run energy consumption height.
Summary of the invention
The present invention is in order to overcome above-mentioned problems of the prior art and shortcoming, a kind of method for preparing fish oligopeptide with fish offal material fermentation coupled film technology is provided, fish oligopeptide comprises Isin glue collagen oligopeptide and fish mixed protein oligopeptide, the method is with the fish offal material after pulverizing, adopt multiple bacteria compound fermentation, the isolation technics that is used in conjunction by three grades of films is again carried out removal of impurities to hydrolyzate, degerming, refining, concentrate and desalination, collect concentrate concentrated further concentrated through circulation low temperature, again after vacuum freeze drying the oligopeptide product, efficiently solve in the original fish oligopeptide production technology cost high, energy consumption is high, product purity is hanged down and the low problem of Product Activity.
The technical solution adopted in the present invention is:
The method for preparing fish oligopeptide with fish offal material fermentation coupled film technology, it is characterized in that carrying out according to the following steps: (1) pulverizes the fish offal material, the percentage by weight that adds water in the entry adjustment fish offal material is 40%～70%, in 115 ℃ of sterilizations 10-30 minute, equitemperature drops to 25～45 ℃, the fermented by mixed bacterium that adds the dry weight 2%～8% that accounts for the fish offal material is regulated the pH value to 6.5-8.0, ferments 36～96 hours; (2) zymotic fluid is after water elution goes out collagen oligopeptide or mixed protein oligopeptide, carries out removal of impurities, degerming, refining, concentrated and desalination that micro-filtration, ultrafiltration, three grades of films of nanofiltration are used in conjunction the liquid that is hydrolyzed after decolouring, coarse filtration; (3) through the concentrate of third level film again after circulation low temperature is further concentrated, carry out again vacuum freeze drying and get Isin glue collagen oligopeptide or fish mixed protein oligopeptide.
In such scheme, described fish offal material is for mixing leftover bits and pieces, single fish scale leftover bits and pieces or single fish-skin leftover bits and pieces, take mix leftover bits and pieces as raw material through fermenting and producing be fish mixed protein oligopeptide, be collagen oligopeptide take single fish scale or single fish-skin as raw material production.
In such scheme, the percentage by weight of water is 40%～60% in the front fish offal material of described fermentation.
In such scheme, the temperature of described fermentation is 28～35 ℃.
In such scheme, described many bacterial classifications refer to two kinds in aspergillus terricola, actinomyces, the bacillus pumilus.
In such scheme, 2%～6% of the dry weight that described fermented bacterium inoculum concentration is the fish offal material.
In such scheme, described fermentation time is 64～80 hours.
The advantage of technical solution of the present invention is:
1, a production line is produced 2 kinds of different products, satisfies the different market demands, has saved the industrial production input cost.
2, adopt multiple bacteria compound fermentation, solve the Current Situation of the low and commercially available protease price costliness of traditional endogenous enzymes hydrolysis efficiency; Fermentation replaces traditional mode of production Isin glue collagen peptide acid-base pretreatment process, avoids widely applying the flushing of the chemical reagent such as acid, alkali and great lot of water resources, has saved resource, has protected environment.
3, the effective multiple proteins component in the aminosal functional areas of this project technology discharges multiple active peptide with biological function, has improved to greatest extent the activity substance content of product.
4, adopt removal of impurities after membrane technology is used for fermentation, refining and concentrated, can obtain high-purity, molecular weight concentrates on the product about 500Da.Simultaneously the volume of polypeptide solution can be reduced to 1/4th of zymotic fluid, greatly reduce the required energy consumption of follow-up drying process, can also accomplish that in addition process water recycles, both reduce water resources consumption, reduce again production waste.
5, adopt one step of film desalination and concentration to finish, cut resin desalination operation, peptide loses problem large and the generation large amount of sewage when having avoided the resin desalination.
6, adopt the concentrated and freeze drying process of low-temperature circulating, replaces that traditional economic benefits and social benefits concentrate, drying process with atomizing.Whole technical process is in normal temperature or cryogenic conditions, the oligopeptide loss of activity of avoiding high temperature to cause, and the activity of product is protected.
Compare with other prior aries, the present invention concentrates on about 500Da with the fish mixed protein oligopeptide molecular weight overwhelming majority of fish mixing waste production, is easy to absorption of human body, and the biologically active high product is without bitter taste, and mouthfeel is joyful; Technology is concentrated more than 70% as the Isin glue collagen oligopeptide molecular weight of raw material production take fish-skin, fish phosphorus and is distributed in about 500Da, product yield (dry fish scale) is more than 34%, collagen peptide purity has good antioxidant effect and activating cell function effect more than 90%.
Multiple bacteria compound fermentation has not only solved the low present situation of endogenous enzymes percent hydrolysis, has more avoided adding the high cost of commercially available protease, also has the effect of directionally hydrolyzing, can hydrolysis has the oligopeptide product than high bioactivity; The film processing procedure can realize the removal of impurities of hydrolyzate, refining, concentrated and desalination, improves the purity of product, and low-temperature circulating is concentrated can further to concentrate the NF membrane concentrate, makes dry treating capacity reduce 3/4, has greatly reduced drying cost; Can realize that by the fine setting zymotechnique two kinds of products of a production line production to satisfy the different market demands, have widened market and reduced the investment in fixed assets cost.
The specific embodiment
Embodiment 1
Get fish and mix leftover bits and pieces 100Kg pulverizing, the percentage by weight that adds water in the entry adjustment fish offal material is 60%, in 115 ℃ of high-temperature sterilizations 20 minutes, equitemperature drops to 30 ℃, access bacillus pumilus and the native bacillus that dwells, bacillus pumilus and the by weight ratio mixing in 1: 1 of native bacillus of dwelling, the bacterial classification amount is 2% of fish mixing leftover bits and pieces dry weight, and adjusting pH value to 8.0, fermented with this understanding 64 hours, and then be warming up to 45 ℃, constant temperature 8 hours, then wash zymotic fluid 2 times, extract oligopeptide, merge water lotion twice, 30min again decolours, filter-cloth filtering, protein extracting ratio are 93%; Hydrolyzate after the coarse filtration is selected the removal of impurities of 10nm microfiltration membranes, degerming, and operating condition is: operating pressure P=0.2MPa, work temperature=50 ℃, flow velocity Q=60L/min; Select molecular cut off 2500Da milipore filter refining, remove most of molecular weight greater than the peptide of 1000Da, see through the oligopeptide that molecular weight is lower than 1000Da, operating condition is: P=0.5MPa, T=40 ℃, Q=30L/min; Select the NF membrane that sees through molecular weight 150～200Da to carry out desalination, concentrated, operating condition is: P=1.5MPa, T=40 ℃, Q=20L/min.Can get the oligomeric Gly-His-Lys of 9.9kg fish mixed protein through the concentrated final vacuum freeze drying of low-temperature circulating again.
Embodiment 2
Fish mixes leftover bits and pieces 100kg and pulverizes, the percentage by weight that adds water in the entry adjustment fish offal material is 70%, in 115 ℃ of high-temperature sterilizations 10 minutes, equitemperature drops to 35 ℃, access bacillus pumilus and actinomyces, bacillus pumilus and actinomyces by weight 1: 1 ratio mix, inoculum concentration is 2% of fish mixing leftover bits and pieces dry weight, and regulate pH value to 7.5, and fermented with this understanding 28 hours, then be warming up to 45 ℃ of constant temperature 8 hours, zymotic fluid water extraction 2 times, water liquid uses active carbon in 50 ℃ of decolourings 30 minutes again, and the coarse filtration rear filtrate is processed through film, and protein extracting ratio is 94%; Film processing procedure and operating condition obtain the oligomeric Gly-His-Lys of 10.1kg fish mixed protein at last with case study on implementation 1.
Embodiment 3
Dry fish scale 80kg pulverizes, the percentage by weight that adds water in the entry adjustment fish offal material is 40%, in 115 ℃ of high-temperature sterilizations 30 minutes, equitemperature drops to 25 ℃, access aspergillus terricola and actinomyces, aspergillus terricola and actinomyces by weight 1: 1 ratio mix, inoculum concentration is 6% of fish scale dry weight, and regulate pH value to 6.5, and fermented with this understanding 72 hours, then be warming up to 45 ℃ of constant temperature 8 hours, zymotic fluid water extraction 2 times, water liquid uses active carbon in 50 ℃ of decolourings 30 minutes again, and the coarse filtration rear filtrate is processed through film, and protein extracting ratio is 92%; Film processing procedure and operating condition obtain the oligomeric Gly-His-Lys of 27.2kg Isin glue collagen at last with case study on implementation 1.
Embodiment 4
Dry fish scale 80kg pulverizes, add entry and adjust the percentage by weight 50% of water in the fish offal material, in 115 ℃ of high-temperature sterilizations 20 minutes, equitemperature drops to 28 ℃, access aspergillus terricola and bacillus pumilus, aspergillus terricola and bacillus pumilus by weight 1: 1 ratio mix, inoculum concentration is 8% of fish scale dry weight, and regulate pH value to 7, and fermented with this understanding 56 hours, then be warming up to 45 ℃ of constant temperature 8 hours, zymotic fluid water extraction 2 times, water liquid uses active carbon in 50 ℃ of decolourings 30 minutes again, and the coarse filtration rear filtrate is processed through film, and protein extracting ratio is 90%; Film processing procedure and operating condition obtain the oligomeric Gly-His-Lys of 26.9kg Isin glue collagen at last with case study on implementation 1.
Embodiment 5
Dried fish-skin 80kg pulverizes, add entry and adjust the percentage by weight 50% of water in the fish offal material, in 115 ℃ of high-temperature sterilizations 20 minutes, equitemperature drops to 35 ℃, access aspergillus terricola and bacillus pumilus, aspergillus terricola and bacillus pumilus by weight 1: 1 ratio mix, inoculum concentration is 5% of fish scale dry weight, and regulate pH value to 7.5, and fermented with this understanding 88 hours, then be warming up to 45 ℃ of constant temperature 8 hours, zymotic fluid water extraction 2 times, water liquid uses active carbon in 50 ℃ of decolourings 30 minutes again, and the coarse filtration rear filtrate is processed through film, and protein extracting ratio is 91%; Film processing procedure and operating condition obtain the oligomeric Gly-His-Lys of 26.9kg Isin glue collagen at last with case study on implementation 1.
Claims (6)
1. the method for preparing fish oligopeptide with fish offal material fermentation coupled film technology, it is characterized in that carrying out according to the following steps: (1) pulverizes the fish offal material, the percentage by weight that adds water in the entry adjustment fish offal material is 40%～70%, in 115 ℃ of sterilizations 10-30 minute, equitemperature drops to 25～45 ℃, the fermented by mixed bacterium that adds the dry weight 2%～8% that accounts for the fish offal material is regulated the pH value to 6.5-8.0, ferments 36～96 hours; (2) zymotic fluid is after water elution goes out collagen oligopeptide or mixed protein oligopeptide, carries out removal of impurities, degerming, refining, concentrated and desalination that micro-filtration, ultrafiltration, three grades of films of nanofiltration are used in conjunction the liquid that is hydrolyzed after decolouring, coarse filtration; (3) through the concentrate of third level film again after further circulation low temperature is concentrated, carry out again vacuum freeze drying and get Isin glue collagen oligopeptide or fish mixed protein oligopeptide; Described mixed bacteria refers to two kinds in aspergillus terricola, actinomyces, the bacillus pumilus.
2. method according to claim 1 is characterized in that: described fish offal material is for mixing leftover bits and pieces, single fish scale leftover bits and pieces or single fish-skin leftover bits and pieces.
3. method according to claim 1 is characterized in that: before the described fermentation in the fish offal material percentage by weight of water be 40%～60%.
4. method according to claim 1, it is characterized in that: the temperature of described fermentation is 28～35 ℃.
5. method according to claim 1 is characterized in that: 2%～6% of the dry weight that described fermented bacterium inoculum concentration is the fish offal material.
6. method according to claim 1, it is characterized in that: described fermentation time is 64～80 hours.
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Families Citing this family (8)
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| CN101974460B (en) * | 2010-09-28 | 2012-05-23 | 中国科学院南海海洋研究所 | Ocean source Bacillus barbaricus SCSIO 02429 and method for preparing squid small peptide by using same |
| TWI442928B (en) * | 2010-11-03 | 2014-07-01 | Food Industry Res & Dev Inst | Preparation and use of fish skin fermentation product |
| CN102199646B (en) * | 2011-03-15 | 2014-08-13 | 广东海洋大学 | Method for preparing collagen peptide with fish skin |
| CN102492761A (en) * | 2011-12-02 | 2012-06-13 | 安徽燕之坊食品有限公司 | Method for preparing active polypeptide from fish pomace |
| CN102517367B (en) * | 2011-12-15 | 2014-04-02 | 河南科技大学 | Method for preparing collagen anti-oxidation peptide by using pig skin |
| CN103211199B (en) * | 2013-05-13 | 2014-09-24 | 武汉梁子湖水产品加工有限公司 | Method for preparing fresh fish essence through processed freshwater fish leftover binary fermentation |
| CN105285312A (en) * | 2014-06-12 | 2016-02-03 | 上海中科国嘉技术转移有限公司 | Preparation method of seafood flavor peptide |
| CN109953176A (en) * | 2017-12-23 | 2019-07-02 | 青岛麦迪尔生物工程有限公司 | A method of peptone is prepared using aquatic products processing leftover bits and pieces |
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