Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Paenibacillus damxungensis sp.nov. CGMCC No.8333 chymosin and preparation method

A technology of Paenibacillus and Bacillus, which is applied in the field of Paenibacillus CGMCC No.8333 chymosin and its preparation, can solve the problems of difficult to meet large-scale industrial production, low recovery rate of chymosin, complicated operation process, etc., and achieve convenient Industrialized operation, simple and effective operation, and the effect of simplifying the preparation process steps

Inactive Publication Date: 2015-09-16
BRIGHT DAIRY & FOOD
View PDF7 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chinese patent application (CN104450655A) discloses the use of ethanol to extract Paenibacillus BD3526 bran fermentation supernatant chymosin, but its operation process is complicated, the recovery rate of gained chymosin is low, it is difficult to meet the needs of large-scale industrial production

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Paenibacillus damxungensis sp.nov. CGMCC No.8333 chymosin and preparation method
  • Paenibacillus damxungensis sp.nov. CGMCC No.8333 chymosin and preparation method
  • Paenibacillus damxungensis sp.nov. CGMCC No.8333 chymosin and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Paenibacillus BD3526 was inoculated in TYC medium and cultured at 30°C for 20 hours to obtain seed liquid. The TYC medium was composed of the following components: casein peptone 15g / L, sucrose 50g / L, yeast extract 5.0g / L, L-cystine 0.2g / L, sodium acetate 20g / L, Na 2 SO 4 0.1g / L, NaCl 1g / L, NaCl 2 HPO 4 12H 2 O 2g / L, NaHCO 3 2g / L, agar 15g / L, and the balance is water.

[0042] Inoculate 4% (v / v) of the seed liquid into a 250mL Erlenmeyer flask containing 50mL of bran medium, ferment and cultivate for 72 hours at 30°C with a shaker shaking speed of 180r / min to obtain BD3526 fermentation broth. The composition of the bran medium is: the mass percentage of wheat bran is 2%, and the balance is water.

[0043] Centrifuge the BD3526 fermentation broth at 4°C and 15,000g for 15 minutes, collect the fermentation supernatant Q1, dilute the fermentation supernatant 3 times with 0.02mol / L, pH 7.0 PBS solution, and measure the fermentation supernatant Q1 curd Enzyme activi...

Embodiment 2

[0050] Paenibacillus BD3526 was inoculated in TYC medium and cultured at 30°C for 20 hours to obtain seed liquid. The TYC medium was composed of the following components: tryptone 15g / L, sucrose 50g / L, yeast extract 5.0g / L, L-cystine 0.2g / L, sodium acetate 20g / L, Na 2 SO 4 0.1g / L, NaCl 1g / L, NaCl 2 HPO 4 12H 2 O 2g / L, NaHCO 3 2g / L, agar 20g / L, and the balance is water.

[0051] Inoculate 2% (v / v) of the seed liquid into a 250mL Erlenmeyer flask containing 50mL of bran medium, ferment and cultivate for 48 hours at 28°C with a shaker at a shaking speed of 300r / min to obtain BD3526 fermentation broth. The composition of the bran medium is: the mass percentage of wheat bran is 1%, and the balance is water.

[0052] Centrifuge the BD3526 fermentation broth at 4°C and 15,000g for 15 minutes, collect the fermentation supernatant Q1, dilute the fermentation supernatant 3 times with 0.02mol / L, pH 7.0 PBS solution, and measure the fermentation supernatant Q1 curd Enzyme activit...

Embodiment 3

[0058] Paenibacillus BD3526 was inoculated in TYC medium and cultured at 28°C for 20 hours to obtain seed liquid. The TYC medium was composed of the following components: casein peptone 15g / L, sucrose 50g / L, yeast extract 5.0g / L, L-cystine 0.2g / L, sodium acetate 20g / L, Na 2 SO 4 0.1g / L, NaCl 1g / L, NaCl 2 HPO 412H 2 O 2g / L, NaHCO 3 2g / L, agar 15g / L, and the balance is water.

[0059] Inoculate 4% (v / v) of the seed liquid into a 250mL Erlenmeyer flask containing 50mL of bran medium, ferment and cultivate for 72 hours at 30°C with a shaker shaking speed of 180r / min to obtain BD3526 fermentation broth. The composition of the bran medium is as follows: the mass percentage of wheat bran is 5%, and the balance is water.

[0060] Centrifuge the BD3526 fermentation broth at 4°C and 15,000g for 15 minutes, collect the fermentation supernatant Q1, dilute the fermentation supernatant 3 times with 0.02mol / L, pH 7.0 PBS solution, and measure the fermentation supernatant Q1 curd Enz...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a paenibacillus damxungensis sp.nov. CGMCC No.8333 chymosin and a preparation method. The method comprises the following steps: (1) centrifuging fermentation broth of which the preservation number is CGMCC No.8333 to obtain fermenting supernate; (2) mixing the fermenting supernate and ammonium sulfate until the saturability of the ammonium sulfate in the fermenting liquid is 50-75% so as to obtain a mixed solution, standing the mixed solution for 0-5 hours at 2-6 DEG C, and centrifugally collecting a precipitate; (3) dissolving the precipitate centrifugally to obtain the supernate. According to the method provided by the invention, the steps of a preparation process of chymosin can be greatly simplified; furthermore, compared with a method of extracting chymosin by using ethanol, the production cost can be greatly saved, the method is convenient and effective to operate, safe and reliable to use, and is beneficial for industrial production.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to Paenibacillus CGMCC No.8333 chymosin and a preparation method. Background technique [0002] Compared with plant-derived and animal-derived chymosin, microbial-derived chymosin has the advantages of low production cost, high biochemical diversity, and easy gene modification. At present, more and more reports indicate that the extracellular proteases of microorganisms have the function of curdling milk, and the chymosin of some microorganisms has achieved large-scale industrial production and is widely used in cheese production. Microorganisms capable of producing extracellular proteases with a certain milk-clotting effect mainly belong to the category of actinomycetes, bacteria, and molds, including the genus Streptomyces, Micromonospora, and Actinomyces madura among the actinomycetes. Rhizopus, Mucor racemosa, Mucor micromyces, Mucor miehei, Mucor fragilis in molds, Phytophthora po...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N9/52C12R1/01
CPCC12N9/6483C12N9/52C12Y304/23004
Inventor 杭锋王钦博刘振民陈卫穆海波王国骄刘沛毅洪青雍靖怡齐晓彦
Owner BRIGHT DAIRY & FOOD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com