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Method for preparing Rhizomucor miehei renninum through liquid fermentation of bran

A technology of chymosin and liquid fermentation of Mucor nigricans is applied in the field of bioengineering, and can solve the problems of high production cost, low enzyme production activity, complicated process and the like, and achieve the effects of low production cost and high activity.

Inactive Publication Date: 2013-06-19
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The enzyme-producing activity of domestic microbial rennet strains is relatively low, and the process is complicated, resulting in high production costs and difficult industrialized large-scale production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Inoculate the preserved mucormycetes species on the prepared PDA slant, incubate at 30°C for 72 hours, and use it when the spores are mature; inject 5mL sterile saline into the mature test tube slant, and use an inoculation loop or a sterile glass Stir with a stick, scrape off the spores of Mucor miehei, oscillate evenly and dilute to 10 6 individual / mL to obtain a spore suspension; add bran to tap water at 10% (W / V), sterilize at 121°C for 10min, and filter through four layers of gauze to obtain a liquid fermentation medium; add the spore suspension to 5% (V / V) V) Insert into the liquid fermentation medium, shake the flask at 30°C and 180r / min for 96h to obtain the fermentation broth; centrifuge the fermentation broth at 5000r / min and 4°C for 15min at low temperature, take the supernatant and pass it through a 0.45um microporous membrane , to obtain the enzyme solution.

Embodiment 2

[0022] Inoculate the preserved mucormycetes species on the prepared PDA slant, incubate at 30°C for 84 hours, and use it when the spores are mature; inject 5mL sterile saline into the mature test tube slant, and use an inoculation loop or a sterile glass Stir with a stick, scrape off the spores of Mucor miehei, oscillate evenly and dilute to 10 6 individual / mL to obtain a spore suspension; add bran to tap water at 10% (W / V), sterilize at 121°C for 10min, and filter through four layers of gauze to obtain a liquid fermentation medium; add the spore suspension to 5% (V / V) V) Insert into the liquid fermentation medium, shake the flask at 30°C and 200r / min for 108h to obtain the fermentation broth; centrifuge the fermentation broth at 5000r / min and 4°C for 15min at low temperature, take the supernatant and pass it through a 0.45um microporous membrane , to obtain the enzyme solution.

Embodiment 3

[0024] Inoculate the preserved mucormycetes species on the prepared PDA slant, incubate at 30°C for 96 hours, and use it when the spores are mature; inject 5mL sterile saline into the mature test tube slant, and use an inoculation loop or a sterile glass Stir with a stick, scrape off the spores of Mucor miehei, oscillate evenly and dilute to 10 6 individual / mL to obtain a spore suspension; add bran to tap water at 10% (W / V), sterilize at 121°C for 10min, and filter through four layers of gauze to obtain a liquid fermentation medium; add the spore suspension to 5% (V / V) V) Insert into the liquid fermentation medium, shake the flask at 30°C and 220r / min for 120h to obtain the fermentation broth; centrifuge the fermentation broth at 5000r / min and 4°C for 15min at low temperature, and take the supernatant and pass it through a 0.45um microporous membrane , to obtain the enzyme solution.

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Abstract

The invention discloses a method for preparing Rhizomucor miehei renninum through liquid fermentation of bran, which comprises the following steps: activating a strain, preparing a spore suspension, preparing a liquid fermentation culture medium, inoculating, performing fermentation culture, and preparing an enzyme liquid. The preparation method disclosed by the invention has the advantages of low raw material cost, simple operating process and short fermentation period, lowers the production cost and is suitable for being popularized in production.

Description

technical field [0001] The invention belongs to the technical field of bioengineering and relates to a method for preparing Mucor miehei rennet by liquid fermentation using bran as a raw material. Background technique [0002] Wheat bran, commonly known as bran and wheat bran, is a by-product obtained after processing flour from wheat seeds. It is mainly the seed coat of wheat, accounting for about 20% of the mass of wheat grains. my country has huge wheat bran resources, with an annual output of about 70-80 million tons. Wheat bran is loose in texture, small in specific gravity and large in surface area. It contains 9.4-17.5% crude protein, 1.7-5.6% crude fat, and is also rich in dietary fiber, vitamins, minerals, oligosaccharides and phenolic compounds. At present, wheat bran is mostly used as feed ingredients or raw materials for vinegar and soy sauce brewing after simple processing, and the utilization rate is low. [0003] Rennet is a key enzyme in the production of c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/58C12R1/785
Inventor 张卫兵艾对元赵炜
Owner GANSU AGRI UNIV
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