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Production method of protease hydrolyzing leftovers of freshwater fish and other fish efficiently

A production method, protease technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, enzymes, etc., can solve the problems of expensive products, achieve the effects of improving permeability, strong elastase activity, and promoting secretion

Active Publication Date: 2010-05-12
JIMEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the production method of elastase is extracted from pig pancreas, and the product price is very expensive, which is not suitable for popularization and application in the food field

Method used

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  • Production method of protease hydrolyzing leftovers of freshwater fish and other fish efficiently

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Experimental program
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Embodiment approach 1

[0017] The invention is a protease production method capable of efficiently hydrolyzing freshwater fish and fish leftovers. The process steps include: (1) strain activation: the culture medium for strain activation consists of: 0.4% beef extract, 0.6% peptone, Yeast extract 0.2%, NaCl 0.5%, agar 2%, moisture 96.3%, pH 7.5; the culture temperature for strain activation is 37°C, and the culture time is 24h; (2) Seed preparation: the medium composition for seed preparation is: 0.4% beef extract, 0.6% peptone, 0.2% yeast extract, 0.5% NaCl, 98.3% water, pH7.5; the preparation of shake flask strains is carried out in a 250ml Erlenmeyer flask, the amount of seed medium is 20ml, and the seed After the medium is prepared, inoculate the newly activated slant lawn into the medium, put it in a shaker, and cultivate it at 180r / m and 32°C for 24h; the preparation of workshop seeds is carried out in the seed tank. The filling coefficient of the culture medium is 0.7, the inoculum size is 1%...

Embodiment approach 2

[0019] The present embodiment is basically the same as the first embodiment, and the difference is: (3) prepare the Bacillus subtilis culture medium, the composition of the culture medium is: glucose 8%, corn steep liquor powder 1%, K 2 HPO 3 0.2%, MgSO 4 ·7H 2 O0.034%, moisture 90.77%; the filling coefficient of the medium is 0.7; after the medium is sterilized and cooled, the inoculation amount of 10% is inserted into the strain, and the fermentation temperature is 37°C, and the dissolved oxygen is 30% ± 5%. , pH 7.4±0.2 and cultured for 16 hours (4) Add 0.1% inducer and 0.06% triton-100, raise the fermentation temperature to 40°C and keep the dissolved oxygen and pH unchanged, and continue to ferment for 16 hours according to the process specification Step (5) extracting elastase and preparing enzyme powder.

Embodiment approach 3

[0021] The present embodiment is basically the same as the first embodiment, the difference is: (3) prepare the Bacillus subtilis medium, the composition of the medium is: glucose 4.5%, corn steep liquor 0.6%, K 2 HPO 3 0.13%, MgSO 4 ·7H 2 O 0.034%, moisture 94.74%; the filling coefficient of the medium is 0.7; when the medium is sterilized and cooled, insert the newly cultivated seeds with an inoculum of 10%, and ferment at a temperature of 37°C and dissolved oxygen of 30%± After culturing for 14 hours under the conditions of 5%, pH 7.4±0.2, add 0.1% inducer and 0.06% triton-100, increase the fermentation temperature to 37°C and keep the dissolved oxygen and pH unchanged, and continue to ferment for 16 hours according to the process Step (5) of the instruction manual extracts elastase and prepares enzyme powder.

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Abstract

The invention discloses a production method of a novel protease. The protease produced by the method has stronger elastase activity and can hydrolyze leftovers of freshwater fish and other fish processing efficiently. The production method of the protease can be summarized as: culturing bacillus subtilis by culture medium without organic nitrogen source till logarithmic phase, adding collagen andtriton-100 extracted from fish scales or fish skin, raising fermentation temperature by 2-3 DEG C, and fermenting continuously for 16h so that elastase activity can reach 650U / mL and total protease activity can reach 2,216U / mL; extracting protease and preparing into powdery enzymic preparation with total protease activity being 18,000U / g and elastase activity being 5,423U / g. Hydrolysis efficiencycan be greatly enhanced by using the protease preparation to hydrolyze leftovers of freshwater fish and other fish processing.

Description

technical field [0001] The invention relates to a method for producing novel protease and its preparation by fermenting Bacillus subtilis, in particular to a protease production method capable of efficiently hydrolyzing freshwater fish and fish waste, and its preparation technology belongs to the technical field of fermentation engineering and biochemical industry , and its application technology involves the fields of medicine, food and cosmetics. Background technique [0002] Enzymatic hydrolysis of low-value fish and fish processing waste to produce high value-added products is an important way for low-value fish and fish comprehensive utilization. Freshwater fish and fish scraps are rich in collagen and less endogenous elastase, and the current commercial protease (alkaline protease, acid protease, compound protease, neutral protease, trypsin, papain, etc.) During hydrolysis, there are generally problems such as low hydrolysis yield and incomplete hydrolysis; therefore,...

Claims

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Application Information

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IPC IPC(8): C12N9/66C12R1/125
Inventor 倪辉蔡慧农肖安风李利君杨远帆
Owner JIMEI UNIV
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