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Method for producing chymosin by aspergillus niger solid fermentation

A technology of solid fermentation and chymosin, which is applied in the field of food biology, can solve the problems of high risk of bacterial contamination in liquid fermentation, difficulty in industrialization, and high cost, and achieve the effects of low hydrolysis capacity, reduced production costs, and low dosage

Inactive Publication Date: 2011-11-23
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

A high-yield and stable chymosin-producing Bacillus subtilis obtained by mutagenesis (Chinese patent CN101665779) had an enzyme activity of only 218.2u / mL through liquid fermentation. The yield was too low and the risk of bacterial infection in liquid fermentation was high. Difficult to achieve industrialization

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  • Method for producing chymosin by aspergillus niger solid fermentation
  • Method for producing chymosin by aspergillus niger solid fermentation
  • Method for producing chymosin by aspergillus niger solid fermentation

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Embodiment 1

[0024] Several different basal media (bran, rice bran, 9:1 mass ratio of bran and flour compound, bran starch compound, rice bran rice flour compound) are selected as carbon source, and the fermentation process (nitrogen source is degreasing) Milk powder, the quality is 1% of the basic medium; 0.7gMgSO is added to every 100ml of water 4 , 0.9gFeSO 4 And 0.1g proline, and use 0.2N HCl to adjust the pH to 6.5~7.0 as acidic nutrient solution; add 15ml of acidic nutrient solution for every 10g solid; the selection range of fermentation temperature is 35~37℃, and the interval of fermentation time is 72~100 hours) Under the same conditions, mix well and sterilize at 121°C for 30 minutes. After cooling, inoculate with Aspergillus niger strain, and carry out solid fermentation. The fermentation temperature is 35~37°C and the fermentation time is 72~100 hours. Water is used as solvent for solid-liquid After extraction, centrifugation and suction filtration, the supernatant was concentrat...

Embodiment 2

[0026] Using a 9:1 mass ratio of bran flour compound as the basic medium carbon source, different nitrogen sources (ammonium nitrate, urea, skimmed milk powder, casein and gelatin) were selected respectively. The nitrogen source quality accounted for 1% of the solid medium quality. %), in its fermentation process (add 0.7gMgSO per 100ml of water 4 , 0.9gFeSO 4 And 0.1g proline, and adjust the pH to 6.5~7.0 with 0.2N HCl, it is acidic nutrient solution; add 15ml acidic nutrient solution for every 10g solid) under the same conditions, mix and sterilize at 121℃ for 30 minutes, and use Aspergillus niger after cooling. Strains were inoculated and solid fermentation was carried out. The fermentation temperature was 35~37℃, and the fermentation time was 72~100 hours. After solid-liquid extraction with water as solvent, centrifugation and suction filtration, the supernatant was concentrated by vacuum rotary evaporation and freeze-dried , To obtain the coarse dry powder of rennet, the re...

Embodiment 3

[0028] To a solid medium prepared with a 9:1 mass ratio of bran flour compound as a basic medium carbon source and 1% skimmed milk powder as a nitrogen source, add an acidic nutrient solution containing inorganic salts: each 100ml water is added separately 0.7gFeSO 4 Or 0.9gMgSO 4 , Add 0.1g proline at the same time, adjust the pH to 6.5-7.0 with 0.2N HCl, add 15ml acidic nutrient solution for every 10g solid. Mix and sterilize at 121°C for 30 minutes. After cooling, inoculate with Aspergillus niger strain and carry out solid fermentation. The fermentation temperature is 35~37°C and the fermentation time is 72~100 hours. Water is used for solid-liquid extraction and centrifugation. , After suction filtration, the supernatant was concentrated by vacuum rotary evaporation and lyophilized to obtain a coarse dry powder of rennet. The rennet activity and protease activity in the Aspergillus niger product image 3 . The experimental results show that Fe 2+ Ions and Mg 2+ The presence...

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Abstract

The invention discloses a method for producing chymosin by aspergillus niger solid fermentation. The method adopts a solid basal medium and comprises the following steps of adding a nitrogen source and an acidic nutrient solution containing one or more inorganic salts and an amino acid into the solid basal medium to wet it, mixing well, carrying out a sterilization process at a temperature of 121 DEG C for 30 minutes, cooling, inoculating the sterilized solid basal medium with an aspergillus niger strain, carrying out solid fermentation at a fermentation temperature of 35 to 37 DEG C for 72 to 100 hours, carrying out solid-liquid extraction through adopting water as a solvent, carrying out centrifugation and extraction filtration, and carrying out the processes of vacuum rotation, evaporation, concentration and freeze-drying on supernate obtained by the previous step to obtain crude dry powder of chymosin. Chymosin which has the advantages of low added amount in raw milk such as milk and the like, rapid curd speed, specific catalytic site and low capacity for non-specific protein hydrolysis is produced through the method.

Description

Technical field [0001] The invention relates to a method for producing rennet by solid fermentation of Aspergillus niger (CICC3.316), and belongs to the field of food biotechnology. Background technique [0002] Rennet is an acid protease, also known as aspartic protease. Since its discovery, it has been widely used in the dairy processing industry. This enzyme is a key enzyme that coagulates milk and produces flavor in cheese production, and it accounts for up to 15% of the output value in the enzyme industry. The traditional source of rennet is the fourth stomach of lactating calves. At present, about 50 million calves need to be slaughtered in the world every year, which still cannot meet the market's increasing demand for cheese products. People are gradually turning their attention to the production of cow rennet. Substitutes. There are many sources of rennet, including animals, plants and microorganisms. Almost all proteolytic enzymes can coagulate milk under appropriate...

Claims

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Application Information

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IPC IPC(8): C12N9/62C12R1/685
Inventor 袁其朋廖亮梁浩黄燕
Owner BEIJING UNIV OF CHEM TECH
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