Method for producing chymosin from paenibacillus damxungensissp.nov. in bran culture medium

A technology of bran culture medium and Paenibacillus, applied in the biological field, can solve the problems of low yield and low yield, and achieve the effects of saving social resources, improving vitality, and good economic and social benefits

Inactive Publication Date: 2014-09-24
BRIGHT DAIRY & FOOD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a production method of microbial s

Method used

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  • Method for producing chymosin from paenibacillus damxungensissp.nov. in bran culture medium
  • Method for producing chymosin from paenibacillus damxungensissp.nov. in bran culture medium

Examples

Experimental program
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Example Embodiment

[0025] Example 1 Preparation of seed liquid

[0026] Take 30mL of TYC liquid culture medium and place them in 100mL conical flasks, seal them with multiple layers (8-12 layers) of gauze and kraft paper, and sterilize them with high-temperature steam at 121°C for 15 minutes;

[0027] Pick a fresh culture of Paenibacillus damxungensis sp.nov. CGMCC No.8333 cultured on TYC solid medium (agar) for 24 hours, inoculate it into TYC liquid medium, and cultivate on a shaker at 30°C for 20 hours to obtain seeds liquid.

Example Embodiment

[0028] Example 2

[0029] The seed solution prepared in Example 1 was inoculated at 5% (volume percentage) of the liquid medium and placed in 20 mL of bran medium (the bran content of the bran medium was 3%, and the rest was deionized water) For fermentation, set a shaker temperature of 30°C, a rotation speed of 300r / min, and shake fermentation for 20 hours.

[0030] After culturing for 20 hours, take 5 mL of fermentation broth and centrifuge at 4°C at 9000 r / min. The supernatant is the crude enzyme solution. The obtained crude enzyme solution was diluted 5 times, and 0.5 mL of the diluted solution was taken to determine the MCA value.

[0031] The experimental results showed that the rennet activity of the crude enzyme liquid cultured with 20 mL of bran culture after 20 mL of bran culture was 4760.71 SU / mL.

Example Embodiment

[0032] Example 3

[0033] The seed solution prepared in Example 1 was inoculated at 5% (volume percentage) of the liquid culture medium and placed in 40 mL bran medium (the bran content of the bran medium was 3%, and the rest was deionized water) For fermentation, set a shaker temperature of 30°C, a rotation speed of 300r / min, and shake fermentation for 20 hours.

[0034] After culturing for 20 hours, take 5 mL of fermentation broth and centrifuge at 4°C at 9000 r / min. The supernatant is the crude enzyme solution. The obtained crude enzyme solution was diluted 5 times, and 0.5 mL of the diluted solution was taken to determine the MCA value.

[0035] The experimental results show that the rennet activity of the crude enzyme liquid culture of 40mL bran culture after the bacillus strain (Paenibacillus damxungensis sp.nov.) CGMCC No.8333 was cultured in shaking fermentation at 30°C for 20 hours was 4578.64 SU / mL.

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Abstract

The invention discloses a method for producing chymosin by fermenting paenibacillus damxungensissp.nov. CGMCC (China General Microbiological Culture Collection Center) No.8333 in a bran culture medium. The method comprises the following steps: inoculating paenibacillus damxungensissp.nov. seed liquid to the bran culture medium, and fermenting at 28-32 DEG C for 16-20 hours to obtain fermenting liquid with chymosin activity, wherein the standard liquid loading volume of a conical flask of 250mL is 20mL to 40mL. Through measurement, highest chymosin activity is 6733.81SU/mL. A paenibacillus damxungensissp.nov. fermenting liquid extract with relatively high chymosin activity can be obtained by virtue of optimized culture conditions, and has wide application prospects in cheese production and processing industry.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for producing chymosin by fermenting Paenibacillus damxungensis sp.nov. CGMCC No.8333 in a bran medium. Background technique [0002] In recent years, the cheese industry has developed rapidly. Curdling is a key step in the production of original cheese. Rennet plays a vital role in this process and is directly related to the quality and yield of cheese. Therefore, as the world's cheese production continues to grow, so does the demand for rennet. The annual demand for curd agent in the world is 25×10 6 L, the output accounts for about 20% of the total enzyme preparations in the world, becoming the largest enzyme preparation. The main function of chymosin is to specifically hydrolyze the κ-casein polypeptide chain in milk to form paracasein and hydrophilic glycomacropeptide. When the casein is hydrolyzed to a certain extent, the calcium ions in the system pass t...

Claims

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Application Information

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IPC IPC(8): C12N9/52C12R1/01
CPCC12N9/52C12Y304/23004
Inventor 杭锋郭本恒王钦博韩瑨刘振民陈卫宋馨吴正钧张灏
Owner BRIGHT DAIRY & FOOD
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