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Preparation method and product of paenibacillus chymosin

A technology of Paenibacillus and Bacillus, applied in the direction of microorganism-based methods, biochemical equipment and methods, hydrolytic enzymes, etc., can solve the problems that chymosin has not yet been obtained, achieve high specific activity, simplify preparation process steps, and reduce production cost effect

Inactive Publication Date: 2015-03-25
BRIGHT DAIRY & FOOD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Chinese patent application (CN 103865842A) discloses that the supernatant of Paenibacillus sp. CGMCC No.8333 strain culture has chymosin activity, but no purified chymosin has been obtained

Method used

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  • Preparation method and product of paenibacillus chymosin
  • Preparation method and product of paenibacillus chymosin
  • Preparation method and product of paenibacillus chymosin

Examples

Experimental program
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Effect test

Embodiment 1

[0027] The extraction method of chymosin in the Paenibacillus fermented liquid of embodiment 1

[0028] The bacterial strain Paenibacillus BD3526 described in the present invention has been preserved in the General Microorganism Center (CGMCC) of China Committee for the Collection of Microorganisms on October 14, 2013. The taxonomic name of the strain is Paenibacillus sp., and the preservation number of the strain is: CGMCC No.8333. For details, please refer to the Chinese patent application (CN 103740618A).

[0029] Paenibacillus BD3526CGMCC No.8333 was fermented and cultivated using bran medium. The composition of the bran medium was: the mass percentage of wheat bran was 2%, the balance was water, the sample volume was 30mL, and the temperature was 30°C. The shaking speed of the bed was 300rpm. After 48 hours of fermentation and cultivation time, the BD3526 fermentation broth was collected; the supernatant fermentation broth was collected after centrifugation at 4°C, 15000g...

Embodiment 2

[0055] The extraction method of chymosin in the Paenibacillus fermented liquid of embodiment 2

[0056] Paenibacillus BD3526 CGMCC No.8333 was fermented and cultivated using bran medium. The composition of the bran medium was: the mass percentage of wheat bran was 5%, the balance was water, the sample volume was 30mL, and the temperature was 30°C. The shaking speed of the shaker was 300rpm. After 48 hours of fermentation and cultivation, the BD3526 fermentation broth was collected; the supernatant fermentation broth was collected after centrifugation at 4°C, 15000g, 20min, and a small amount of the supernatant fermentation broth was diluted 5 times with PBS, and Measure the enzyme activity (SU / mg) and protein content (mg / mL) of the fermentation supernatant; mix 95% ethanol with the supernatant fermentation broth at a volume ratio of 4:1 (V / V), mix well and then store at 4°C Let stand for 5h.

[0057] Centrifuge at 15,000g at 4°C for 15min to collect alcohol-precipitated subst...

Embodiment 3

[0061] The extraction method of chymosin in the Paenibacillus fermented liquid of embodiment 3

[0062] Paenibacillus BD3526CGMCC No.8333 was fermented and cultivated using bran medium. The composition of the bran medium was: the mass percentage of wheat bran was 3%, the balance was water, the sample volume was 30mL, the temperature was 30°C, and the The shaking speed of the bed was 300rpm. After 48 hours of fermentation and cultivation time, the BD3526 fermentation broth was collected; the supernatant fermentation broth was collected after centrifugation at 4°C, 15000g, 20min, and a small amount of the supernatant fermentation broth was diluted 5 times with PBS, and measured Enzyme activity (SU / mg) and protein content (mg / mL) of the fermentation supernatant; mix 95% ethanol with the supernatant fermentation broth at a volume ratio of 4:1 (V / V), mix well and then statically place at 4°C set for 1h.

[0063] Centrifuge at 15,000g at 4°C for 15min to collect alcohol-precipitate...

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Abstract

The invention discloses a preparation method and a product of paenibacillus chymosin. The preparation method comprises the following steps: (1) inoculating paenibacillus sp. in a culture medium for culturing paenibacillus sp. to carry out oscillatory fermentation culture and centrifuging the obtained fermentation liquor to get the liquid supernatant; (2) mixing the obtained liquid supernatant with ethyl alcohol and centrifuging to obtain the precipitate; and (3) dissolving the obtained precipitate by water or PBS, centrifuging to get the liquid supernatant and carrying out freeze-drying to obtain the paenibacillus chymosin. The preparation method is capable of greatly simplifying the preparation steps of the chymosin and effectively reducing the production cost of the chymosin under the premise of improving the specific activity and the recovery rate of the chymosin. The chymosin prepared by the preparation method has relatively high specific activity and is capable of sufficiently satisfying the industrial production demand of the chymosin.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method of Paenibacillus chymosin and a product thereof. Background technique [0002] Rennet is a key enzyme in cheese production. It plays a very important role in the formation of cheese texture and unique flavor. In the international market, rennet from animals accounts for about 70%, and microorganisms account for 30%. With the continuous development of the cheese industry, rennet from animal sources is far from meeting the production needs. Researchers from various countries are constantly looking for new products. source. At present, it is found that microorganisms that can produce a certain amount of rennet are mainly actinomycetes, bacteria and molds, including actinomycetes Streptomyces, Micromonospora, Actinomyces madura, Rhizopus, Racemosa Mucor, Mucor micromus, Mucor miehei, Phytophthora pombe, Parasitic Intratheca, Mucor fragilis and other hi...

Claims

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Application Information

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IPC IPC(8): C12N9/52C12R1/01
CPCC12N9/52
Inventor 杭锋王钦博郭本恒刘振民吴正钧韩瑨穆海菠于华宁朱军伟
Owner BRIGHT DAIRY & FOOD
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