CN104313093A - Method for preparing peptone by utilizing fish processing wastes - Google Patents
Method for preparing peptone by utilizing fish processing wastes Download PDFInfo
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- CN104313093A CN104313093A CN201410549672.8A CN201410549672A CN104313093A CN 104313093 A CN104313093 A CN 104313093A CN 201410549672 A CN201410549672 A CN 201410549672A CN 104313093 A CN104313093 A CN 104313093A
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Abstract
The invention discloses a method for preparing peptone by utilizing fish processing wastes, belonging to the technical field of biology. The method is characterized in that the raw materials used for preparing peptone are fish processing wastes; the raw materials are minced and degreased to obtain minced fillet pulp; a certain amount of protease is added; protease enzymolysis procedures are controlled by applying size-exclusion high performance liquid chromatography (SEC-HPLC); membrane separation is further used for desalination and concentration, and the powder obtained after spray drying is fish peptone. The method has the beneficial effects that the fish processing wastes can be fully utilized by adopting the method to prepare high-quality fish peptone; the method has the advantages of retention of full amino acid composition, high content of amino nitrogen, low protein molecular weight, low ash content, and the like; the product can serve as health food, a nutrient enhancer, a food additive, a feed additive, a medium in the fermentation industry, and the like.
Description
Technical field
The invention belongs to biological technical field, be specifically related to a kind of method utilizing fish processing fent to prepare peptone.
Background technology
Peptone be protein through enzyme, acid or basic hydrolysis obtain a kind of by showing by the moon, the water soluble mixt that forms of peptone, peptide and amino acid, be the Major Nutrient nitrogenous source cultivating most of microbe.The raw material that tradition prepares peptone has a variety of, as poultry blood, poultry bone, fish meal and glue slag etc.Current research shows, in peptone products prepared by different material, the enzymolysis product of fish protein resource has the features such as protein content is high, good water solubility, polypeptide and total free aminoacids are abundant, and therefore fish peptone can as the optimum protein peptone in fermentation industry and laboratory selected by microorganism culturing.The Fisheries Development of China is rapid, but less for the exploitation of fish processing fent, major part for processing feed, fish meal or being abandoned by as waste, the lower and contaminate environment of added value.Develop the albumen in fish processing fent, not only can improve the value added of fishery processing, expand the raw material sources of peptone; And effectively can solve environmental issue, there is good economic benefit and social benefit.
The method that tradition prepares peptone has acid-hydrolysis method, alkali hydrolysis method and enzyme hydrolysis method.Acid-hydrolysis method generally adopts the acid reagents such as hydrochloric acid, sulfuric acid and phosphoric acid as hydrolytic reagent.Acid hydrolysis requires high to equipment pipeline; Destructible tryptophane, product colour is comparatively dark, molecular weight distribution is wide, ash oontent is high; Technique needs consider disacidify, decolouring problem.Alkali hydrolysis method adopts the alkaline reagentss such as rare NaOH, ammoniacal liquor, volatile salt and liming as hydrolytic reagent usually.Much amino acid is all subject to destruction in various degree, product generation racemization, and its product is D-type and the amino acid whose mixture of L-type.Enzyme hydrolysis method mild condition, does not destroy amino acid, racemization does not occur.
Chinese patent CN1418562A, CN1086965A, CN1379019, CN1043934 and CN101538601A etc. patent describes the method that enzyme process prepares peptone.These methods all do not relate to raw material--fish processing fent.Protease hydrolysis complex process, raw material sources, protease species and concentration, enzymatic hydrolysis condition (temperature, time and pH) etc. all can affect the quality of final peptone products, above-mentioned technique cannot carry out the precise hard_drawn tuhes of process, thus causes product molecular weight distribution instability, ammonia-nitrogen content not high.At present, most enzyme solution does not relate to desalination program, thus causes product ash content higher; In addition, some method adopts high temperature to concentrate, and causes protein heat denaturation, affects end product quality.
Summary of the invention
The object of the invention is to provide a kind of fish processing fent to prepare the method for high-quality peptone.
For achieving the above object, the present invention adopts following technical scheme:
A kind of method utilizing fish processing fent to prepare peptone, with fish processing fent for raw material, raw material obtains fish gruel slurry through rubbing, degreasing, add protease hydrolyzed, application molecular sieve high-efficient liquid phase chromatography SEC-HPLC carries out detection and analyzes, and enzymolysis terminal point control protein molecular weight distributes, and below 3000Da protein molecule content is higher than 90%, carry out desalination with membrane sepn further and concentrate, the powder after spraying dry and peptone.
Described fish processing fent derive from comprise the gruel that cures fish, tankage that the production line of fillet and canned fish produces.Comprise raw fish, fish head, fish-skin, fish-bone, fish muscle and fish tail.
Described protease hydrolyzed adopts stomach en-, trypsinase, papoid, subtilisin or kethepsin enzymolysis; Adopt single stage method to carry out enzymolysis, or adopt two step method to carry out enzymolysis.
Described single stage method carries out enzymolysis, and it is 0.2-3.0% that the addition of enzyme accounts for fish gruel slurry weight ratio, adjust ph to 1.5 ~ 8.0, and temperature of reaction is 20 ~ 55 DEG C, enzymolysis time 6-24 hour; Described two step method carries out enzymolysis, and the first step enzymolysis adopts stomach en-or papoid, and the addition of enzyme accounts for fish gruel slurry weight ratio 0.1-2.0%, adjust ph to 1.5 ~ 5.0, and temperature of reaction is 20 ~ 55 DEG C, enzymolysis time 3-12 hour; Second step enzymolysis adopts trypsinase or subtilisin, and it is 0.01-2.0% that the addition of enzyme accounts for fish gruel slurry weight ratio, and adjust ph to 1.5 ~ 8.0, temperature of reaction is 20 ~ 55 DEG C, enzymolysis time 5-12 hour.
Application molecular sieve high-efficient liquid phase chromatography SEC-HPLC carries out detection and analyzes: in enzymolysis process, SEC-HPLC analysis is carried out in every 30-60min sampling, adopts SEC-120 gel column to analyze, moving phase: PBS damping fluid; Determined wavelength: 220nm; Flow velocity: 0.35mL/min, isocratic elution, sampling volume 10 μ L, enzymolysis terminal point control protein molecular weight distributes, and below 3000Da protein molecule content is higher than 90%.
Described PBS damping fluid is 0.1M PBS damping fluid, pH6.98.
Described membrane sepn adopts interception to be the membrane module of 100-1000Da.
Adopt method of the present invention to prepare fish peptone and can make full use of fish processing fent, have amino acid composition and retain the advantages such as complete, amino nitrogen content is high, protein molecular weight is little, ash content is low, the product of production can as protective foods, nutrition-fortifying agent, foodstuff additive, fodder additives and fermentation industry substratum etc.
Accompanying drawing explanation
fig. 1 is the molecular weight distribution of fish peptone MFZU01 and fish peptone MFZU02: (A) fish peptone FZU01 molecular weight distribution (SEC-HPLC spectrogram); (B) fish peptone FZU02 molecular weight distribution (SEC-HPLC spectrogram).
Embodiment
Fish processing fent (comprising raw fish, fish head, fish-skin, fish-bone, fish muscle and fish tail etc.), obtains fish gruel slurry through rubbing, degreasing pre-treatment.
Add a certain amount of proteolytic enzyme to be hydrolyzed, single stage method is adopted to carry out enzymic hydrolysis, or adopt two step method to carry out enzymic hydrolysis, molecular sieve high-efficient liquid phase chromatography (SEC-HPLC) controls protease hydrolyzed program, in enzymolysis process, SEC-HPLC analysis is carried out in every 30-60min sampling, adopts SEC-120 gel column to analyze.Moving phase: PBS damping fluid (0.1M, pH6.98); Determined wavelength: 220nm; Flow velocity: 0.35mL/min, isocratic elution, sampling volume 10 μ L.Enzymolysis terminal point control protein molecular weight distributes, and higher than 90%(SEC-HPLC analysis of spectra as shown in Figure 1, molecular weight distribution result is as shown in table 1 for below 3000Da protein molecule content).Single stage method carries out enzymic hydrolysis, and it is 0.2-3.0% that the addition of enzyme accounts for fish gruel slurry weight ratio, adjust ph to 1.5 ~ 8.0, and temperature of reaction is 20 ~ 55 DEG C, enzymolysis time 6-24 hour; Two step method carries out enzymolysis, and the first step enzymolysis adopts stomach en-, and the addition of enzyme accounts for fish gruel slurry weight ratio 0.1-2.0%, adjust ph to 1.5 ~ 5.0, and temperature of reaction is 20 ~ 55 DEG C, enzymolysis time 3-12 hour; Second step enzymolysis adopts trypsinase or subtilisin, and it is 0.01-2.0% that the addition of enzyme accounts for fish gruel slurry weight ratio, and adjust ph to 1.5 ~ 8.0, temperature of reaction is 20 ~ 55 DEG C, enzymolysis time 5-12 hour.
Employing interception is that the membrane module of 100-1000Da carries out desalination and concentrates, the powder after spraying dry and fish peptone.
By several specific embodiment of the present invention given below, further describe the present invention, but they not limitation of the invention:
embodiment 1
Get raw fish 100 kilograms, rub with mincer, add 300 liters of No. 60 solvent oil skimming treatments 3 times, obtain degreasing fish gruel slurry.Add 400L water, the acetum with 10% regulates pH, makes final pH reach 3.5.Adopt two step enzymolysis process.The first step enzymolysis adopts papoid, and the addition of enzyme is 0.8%, adjust ph to 3.0, and temperature of reaction is 40 DEG C, enzymolysis time 8 hours; Second step enzymolysis adopts trypsinase, and the addition of enzyme is 0.7%, adjust ph to 4.5, and temperature of reaction is 40 DEG C, enzymolysis time 12 hours.Enzymolysis solution detects through SEC-HPLC, and lower than 3000Kda(SEC-HPLC analysis of spectra as shown in Figure 1A, molecular weight distribution result is as shown in table 1 fish peptone FZU01 for protein molecular weight more than 90%), stop enzyme reaction.Enzymolysis product is separated through high-speed and continuous centrifuge, collects supernatant liquor, i.e. fish and water solution peptone solution.Adopt membrane separation unit (250Da film) to process above-mentioned material, RO water washing 800L, fish and water solution peptone solution is finally concentrated into 200L for spraying dry.The fish peptone powder collected is analyzed, amino nitrogen 5.2%, nitrogen content 14.7%, ash content 1.6% with reference to " the main raw and auxiliary material quality control standard of Products in China ".
embodiment 2
Get the rotten processing fent 100 kilograms (containing fish head, fish-skin, fish-bone, fish muscle and fish tail etc.) of fish, rub with mincer, add 300 liters of No. 60 solvent oil skimming treatments 3 times, obtain degreasing fish gruel slurry.Add 400L water, regulate pH with 1N hydrochloric acid soln, make final pH reach 3.5.Add the proteolytic enzyme (stomach en-, trypsinase, papoid, subtilisin or kethepsin etc.) of 1.0%, 35 DEG C, under abundant agitation condition, enzymolysis 6h.Enzymolysis solution detects through SEC-HPLC, and lower than 3000Kda(SEC-HPLC analysis of spectra as shown in Figure 1B, molecular weight distribution result is as shown in table 1 fish peptone FZU02 for protein molecular weight more than 90%), stop enzyme reaction.Enzymolysis product is separated through high-speed and continuous centrifuge, collects supernatant liquor, i.e. fish and water solution peptone solution.Adopt membrane separation unit (250Da film) to process above-mentioned material, RO water washing 800L, fish and water solution peptone solution is finally concentrated into 200L for spraying dry.The fish peptone powder collected is analyzed, amino nitrogen 5.8%, nitrogen content 14.6%, ash content 1.9% with reference to " the main raw and auxiliary material quality control standard of Products in China ".
Table 1 peptone molecular weight distribution data
Claims (8)
1. the method utilizing fish processing fent to prepare peptone, it is characterized in that: described method with fish processing fent for raw material, raw material obtains fish gruel slurry through rubbing, degreasing, add protease hydrolyzed, application molecular sieve high-efficient liquid phase chromatography SEC-HPLC carries out detection and analyzes, and enzymolysis terminal point control protein molecular weight distributes, and below 3000Da protein molecule content is higher than 90%, carry out desalination with membrane sepn further and concentrate, the powder after spraying dry and peptone.
2. a kind of method utilizing fish processing fent to prepare peptone according to claim 1, is characterized in that: described fish processing fent derive from comprise the gruel that cures fish, tankage that the production line of fillet and canned fish produces.
3. a kind of method utilizing fish processing fent to prepare peptone according to claim 2, is characterized in that: described tankage comprise raw fish, fish head, fish-skin, fish-bone, fish muscle and fish tail.
4. a kind of method utilizing fish processing fent to prepare peptone according to claim 1, is characterized in that: described protease hydrolyzed adopts stomach en-, trypsinase, papoid, subtilisin or kethepsin enzymolysis; Adopt single stage method to carry out enzymolysis, or adopt two step method to carry out enzymolysis.
5. a kind of method utilizing fish processing fent to prepare peptone according to claim 4, it is characterized in that: described single stage method carries out enzymolysis, it is 0.2-3.0% that the addition of enzyme accounts for fish gruel slurry weight ratio, adjust ph to 1.5 ~ 8.0, temperature of reaction is 20 ~ 55 DEG C, enzymolysis time 6-24 hour; Described two step method carries out enzymolysis, and the first step enzymolysis adopts stomach en-or papoid, and the addition of enzyme accounts for fish gruel slurry weight ratio 0.1-2.0%, adjust ph to 1.5 ~ 5.0, and temperature of reaction is 20 ~ 55 DEG C, enzymolysis time 3-12 hour; Second step enzymolysis adopts trypsinase or subtilisin, and it is 0.01-2.0% that the addition of enzyme accounts for fish gruel slurry weight ratio, and adjust ph to 1.5 ~ 8.0, temperature of reaction is 20 ~ 55 DEG C, enzymolysis time 5-12 hour.
6. a kind of method utilizing fish processing fent to prepare peptone according to claim 1, it is characterized in that: application molecular sieve high-efficient liquid phase chromatography SEC-HPLC carries out detection and analyzes: in enzymolysis process, SEC-HPLC analysis is carried out in every 30-60min sampling, SEC-120 gel column is adopted to analyze, moving phase: PBS damping fluid; Determined wavelength: 220nm; Flow velocity: 0.35mL/min, isocratic elution, sampling volume 10 μ L, enzymolysis terminal point control protein molecular weight distributes, and below 3000Da protein molecule content is higher than 90%.
7. a kind of method utilizing fish processing fent to prepare peptone according to claim 6, is characterized in that: described PBS damping fluid is 0.1M PBS damping fluid, pH6.98.
8. a kind of method utilizing fish processing fent to prepare peptone according to claim 1, is characterized in that: described membrane sepn adopts interception to be the membrane module of 100-1000Da.
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Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105734100A (en) * | 2016-02-26 | 2016-07-06 | 浙江肽鲜乐生物技术有限公司 | Process for producing fish peptone from fish-soluble slurry |
| CN106805252A (en) * | 2017-01-22 | 2017-06-09 | 福州大学 | A kind of fish small peptide and its application in nutrition fortifier |
| CN106834401A (en) * | 2017-01-23 | 2017-06-13 | 舟山福氏食品科技有限公司 | A kind of small band fish peptone production technology |
| CN107227331A (en) * | 2017-07-11 | 2017-10-03 | 浙江丰宇海洋生物制品有限公司 | A kind of method of the molten slurry production protein small peptide of utilization fish |
| CN107226855A (en) * | 2017-07-11 | 2017-10-03 | 浙江丰宇海洋生物制品有限公司 | A kind of peptone of the molten slurry production of utilization fish |
| CN107227332A (en) * | 2017-07-11 | 2017-10-03 | 浙江丰宇海洋生物制品有限公司 | A kind of method of the molten slurry production peptone of utilization fish |
| CN107279991A (en) * | 2017-06-01 | 2017-10-24 | 慈溪创璞食品科技有限公司 | A kind of production method of natural seafood delights nutrient |
| CN107361194A (en) * | 2017-06-01 | 2017-11-21 | 慈溪创璞食品科技有限公司 | A kind of processing method of the molten slurry of biological enzymolysis fish |
| CN107513548A (en) * | 2017-10-31 | 2017-12-26 | 荣成市日鑫水产有限公司 | A kind of method that peptone is prepared using fish meal pressed liquor |
| CN109517847A (en) * | 2018-12-04 | 2019-03-26 | 张白羽 | A method of utilizing aquatic products waste fermenting and preparing biological surfactant |
| CN111334554A (en) * | 2019-07-16 | 2020-06-26 | 烟台市华昕生物医药科技有限公司 | Salmon oligopeptide and preparation method thereof |
| CN113951366A (en) * | 2021-10-26 | 2022-01-21 | 舟山福氏食品科技有限公司 | Processing method for preparing biological enzymolysis protein by using squid leftovers |
| CN114292309A (en) * | 2021-12-24 | 2022-04-08 | 百洋产业投资集团股份有限公司 | Method for producing peptone powder from fish processing by-products |
| CN115975885A (en) * | 2022-12-23 | 2023-04-18 | 中国农业大学 | Preparation of akkermansia muciniphila culture medium by using fish processing byproducts as nitrogen source |
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Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105734100A (en) * | 2016-02-26 | 2016-07-06 | 浙江肽鲜乐生物技术有限公司 | Process for producing fish peptone from fish-soluble slurry |
| CN106805252A (en) * | 2017-01-22 | 2017-06-09 | 福州大学 | A kind of fish small peptide and its application in nutrition fortifier |
| CN106834401A (en) * | 2017-01-23 | 2017-06-13 | 舟山福氏食品科技有限公司 | A kind of small band fish peptone production technology |
| CN107279991A (en) * | 2017-06-01 | 2017-10-24 | 慈溪创璞食品科技有限公司 | A kind of production method of natural seafood delights nutrient |
| CN107361194A (en) * | 2017-06-01 | 2017-11-21 | 慈溪创璞食品科技有限公司 | A kind of processing method of the molten slurry of biological enzymolysis fish |
| CN107227331A (en) * | 2017-07-11 | 2017-10-03 | 浙江丰宇海洋生物制品有限公司 | A kind of method of the molten slurry production protein small peptide of utilization fish |
| CN107226855A (en) * | 2017-07-11 | 2017-10-03 | 浙江丰宇海洋生物制品有限公司 | A kind of peptone of the molten slurry production of utilization fish |
| CN107227332A (en) * | 2017-07-11 | 2017-10-03 | 浙江丰宇海洋生物制品有限公司 | A kind of method of the molten slurry production peptone of utilization fish |
| CN107513548A (en) * | 2017-10-31 | 2017-12-26 | 荣成市日鑫水产有限公司 | A kind of method that peptone is prepared using fish meal pressed liquor |
| CN107513548B (en) * | 2017-10-31 | 2021-01-22 | 荣成市日鑫水产有限公司 | Method for preparing peptone by using fish meal squeezed liquid |
| CN109517847A (en) * | 2018-12-04 | 2019-03-26 | 张白羽 | A method of utilizing aquatic products waste fermenting and preparing biological surfactant |
| CN111334554A (en) * | 2019-07-16 | 2020-06-26 | 烟台市华昕生物医药科技有限公司 | Salmon oligopeptide and preparation method thereof |
| CN113951366A (en) * | 2021-10-26 | 2022-01-21 | 舟山福氏食品科技有限公司 | Processing method for preparing biological enzymolysis protein by using squid leftovers |
| CN114292309A (en) * | 2021-12-24 | 2022-04-08 | 百洋产业投资集团股份有限公司 | Method for producing peptone powder from fish processing by-products |
| CN115975885A (en) * | 2022-12-23 | 2023-04-18 | 中国农业大学 | Preparation of akkermansia muciniphila culture medium by using fish processing byproducts as nitrogen source |
| CN115975885B (en) * | 2022-12-23 | 2024-07-16 | 中国农业大学 | Preparation of mucin-philin Acremonium culture medium by using fish processing byproducts as nitrogen source |
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Application publication date: 20150128 |