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Method for preparing fermented glutinous rice Chinese yeast microorganism sourced chymosin

A technology of glutinous rice wine and chymosin, applied in the direction of hydrolytic enzymes, etc., can solve the problems of strain limitation, incoordination, long time growth cycle, etc., and achieve the effects of simple operation, improvement of vitality, and saving of raw materials

Active Publication Date: 2008-08-27
上海瀚厨餐饮管理有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since rennet is a key enzyme in cheese production, with the continuous development of the cheese industry in the world, the method of slaughtering 50 million calves every year to obtain rennet still cannot meet the production needs, and is also very different from the development of modern industry. coordination
Therefore, researchers are constantly looking for new sources, mainly including animal rennet, plant rennet and microbial source rennet, but these sources of rennet have certain limitations
Animal-based rennet is source-restricted, and cheese made from animal-based rennet does not meet the needs of vegans
Although plant-based rennet has a wide range of sources, it is difficult to develop due to constraints such as time, region, and long growth cycle.
The use of microorganisms to produce chymosin is the most promising development direction at present. The growth cycle of microorganisms is short, the output is large, and it is less restricted by climate, region, and time. But at the same time, there are also other non-enzymatic metabolites produced by microorganisms. Generally, when microbial enzymes become commercial products, they must be tested for toxicity, carcinogenicity and skin allergy, so the strains that are actually used for industrial production of chymosin are very limited.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Add 5.0g of glutinous rice flour and 50mL of water to ten 250mL shake flasks, shake well, sterilize at 115°C for 20 minutes, cool to about 30°C at room temperature, add 0.016g of wine medicine at 30°C, and shake at 140 rpm After 50 hours, it was glutinous rice wine. The protein content in the glutinous rice wine was measured to be 1080.0 mg, and the curd activity was 21.89 U / mg.

[0016] Combine the glutinous rice wine in each shaker flask, centrifuge at 3500 rpm for 10 min at 4°C, and take the supernatant. Slowly add 113.0g of dried and ground ammonium sulfate powder into it, let stand at 4°C for 2 hours, centrifuge (7000 rpm, 15min, 4°C), take the supernatant, continue to add 167.5g of dried and finely ground Grind fine ammonium sulfate powder, let stand at 4°C for 2 hours, centrifuge (8000 rpm, 15min, 4°C), dissolve the precipitate in 0.005mol / L, pH6.28 phosphate buffer, and dialyze at 4°C for 48 hours, That is the crude rennet solution, the protein content in the c...

Embodiment 2

[0019] Add 5.0g of glutinous rice flour and 50mL of water to ten 250mL shake flasks, shake well, sterilize at 115°C for 20 minutes, cool to about 30°C at room temperature, add 0.016g of wine medicine at 30°C, and shake at 140 rpm After 52 hours, it was glutinous rice wine, and the protein content in the glutinous rice wine was measured to be 1135.4 mg, and the curd activity was 23.66 U / mg.

[0020] Combine the glutinous rice wine in each shaker flask, centrifuge at 3500 rpm for 10 min at 4°C, and take the supernatant. Slowly add 113.0g of dried and ground ammonium sulfate powder into it, let stand at 4°C for 2 hours, centrifuge (7000 rpm, 15min, 4°C), take the supernatant, continue to add 167.5g of dried and finely ground Finely ground ammonium sulfate powder, let stand at 4°C for 2 hours, centrifuge (8000 rpm, 15min, 4°C), dissolve the precipitate in 0.005M, pH6.28 phosphate buffer, and dialyze at 4°C for 48 hours, that is The crude rennet solution has a protein content of 6...

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Abstract

The invention relates to a method for preparing wine drug microbial origin chymosin of glutinous rice wine. The method of the invention comprises: utilizing glutinous rice flour as culture medium, producing the glutinous rice wine through fermenting wine drug, adopting an ammonium sulfate graded precipitation method to extract chymosin from the glutinous rice wine, further utilizing Sephadex G-100 and DEAE-Sephadex A-50 to purify, and obtaining the chymosin with high purity. The method is used to produce the chymosin, sources of raw materials are wide, the cost is low, the operation is simple, the fermentation period is short, the chymosin which is produced has high activity, the milk-clotting activity after purifying can reach 4360+-50U / mg, and the protein recovery is 16.42+-1.98%.

Description

【Technical field】 [0001] The invention belongs to the technical field of preparation of biological preparations, and in particular relates to a preparation method of rennet derived from microbial sources of glutinous rice wine medicine. 【Background technique】 [0002] Chymosin (chymosin, EC3.4.23.4) is an aspartic protease found in the stomach of unweaned calves, and its main biological function is the limited shearing of Phe105 in κ-casein of casein The peptide bond linked by -Met106 leads to milk coagulation, so it is widely used in cheese manufacturing and becomes an important enzyme preparation in the food field, and its output value accounts for 15% of the world's enzyme preparation. [0003] Rennet is traditionally prepared by leaching rennet from the fourth stomach of calves 10-30 days old after slaughter. Since rennet is a key enzyme in cheese production, with the continuous development of the cheese industry in the world, the method of slaughtering 50 million calve...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/52
Inventor 王艳萍程巧玲张阳
Owner 上海瀚厨餐饮管理有限公司
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