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Low-temperature alkaline proteinase marine bacteria strain, low-temperature alkaline proteinase and production method thereof

A low-temperature protease and production method technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve problems such as large differences and differences in enzyme characteristics, and achieve high enzyme activity, high and low temperature stability, and convenience Collection of refined effects

Inactive Publication Date: 2011-07-13
ZHEJIANG WANLI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But in general, there are large differences between the current bacterial strains at home and abroad, and there are differences in the enzyme characteristics of the bacteria produced, including optimal action conditions, thermal stability, and hydrolysis characteristics for substrates, etc.

Method used

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  • Low-temperature alkaline proteinase marine bacteria strain, low-temperature alkaline proteinase and production method thereof
  • Low-temperature alkaline proteinase marine bacteria strain, low-temperature alkaline proteinase and production method thereof
  • Low-temperature alkaline proteinase marine bacteria strain, low-temperature alkaline proteinase and production method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Activation culture of low temperature protease producing bacteria (B8453013)

[0044] The low-temperature protease-producing bacteria B8453013 was inoculated on the surface of 2216 medium, and cultured at 15°C for 3 days for rejuvenation. Then inoculate it in the 2216 medium containing 0.1wt% skimmed milk powder for screening, cultivate at 15°C for 3 days to produce an obvious transparent circle around the colony, and pick the strain with the largest diameter ratio of the transparent circle to the colony for subsequent fermentation strain. The composition of 2216 medium is by weight: peptone 0.5%, yeast extract 0.1%, NaCl 11.738%, KCl 0.332%, KBr 0.048%, MgCl 2 ·6H 2 O 5.305%, SrCl 2 6H2O 0.020%, CaCl 2 2H 2 O 0.7345%, Na 2 SO 4 1.9585%, NaHCO 3 0.096%, H 3 BO 3 0.013%, (NH 4 ) 2 SO 4 0.5%, deionized water, pH9.0, 0.1Mpa steam sterilization for 15 minutes.

Embodiment 2

[0045] Embodiment 2: the fermentation culture-I of low-temperature protease producing bacteria (B8453013)

[0046] In this example, the fermentation strain obtained by screening was directly inoculated into the fermentation medium for cultivation. Specifically:

[0047] Inoculate the slant surface of the fermentation strain into the fermentation medium, fill each 250mL shake bottle with 100mL of fermentation medium, the culture conditions are: 15°C, 150 rpm, 72 hours of fermentation, and the enzyme activity reaches 97U / mL. The components of the fermentation medium are by weight: 0.5% casein, 0.5% sodium citrate, 0.5% peptone, 0.1% yeast extract, 11.738% NaCl, 0.332% KCl, 0.048% KBr, MgCl 2 ·6H 2 O 5.305%, SrCl 2 6H2O 0.020%, CaCl 2 2H 2 O 0.7345%, Na 2 SO 4 1.9585%, NaHCO 3 0.096%, H 3 BO 3 0.013%, (NH 4 ) 2 SO 4 0.5%, Tween-800.05%, deionized water, pH9.0, 0.1Mpa steam sterilization for 15 minutes.

[0048] Add ammonium sulfate to the obtained protease liquid...

Embodiment 3

[0049] Embodiment 3: Fermentation culture-II of low temperature protease producing bacteria (B8453013)

[0050] In this embodiment, the strains for fermentation obtained by screening are first subjected to liquid proliferation culture, and then inoculated in the fermentation medium for fermentation. Specifically:

[0051] Inoculate the slant surface of the low-temperature protease-producing strain in 2216 medium for proliferation at 15° C. and 150 rpm. After culturing for 24 hours, the culture is added to the fermentation medium with 15% inoculum size for fermentation. The composition of 2216 medium is the same as in Example 1. The components of the fermentation medium and the sterilization method are the same as the components of the fermentation medium and the sterilization method in Example 2. The fermentation culture is 50 mL of fermentation medium per 250 mL shake bottle, the culture conditions are: 15°C, 200 rpm, 72 hours of fermentation culture, and the enzyme activit...

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Abstract

The invention discloses a marine Pseudoalteromonas sp. B8453013 (strain collection number CGMCC No.3281) of low-temperature proteinase, a production method for producing low-temperature proteinase from the strain and a low-temperature proteinase. A system method for strain collection, rejuvenation and seed selection is built by alternating Pseudoalteromonas sp. B8453013; the method for producing the low-temperature proteinase is built by culture medium optimization screening and the optimized control of fermentation technology conditions; and the produced low-temperature proteinase has the advantages of good low-temperature activity and high enzyme activity. The low-temperature proteinase can be widely applied to industries, such as detergents, feed, fur, food processing and the like.

Description

technical field [0001] The invention relates to the field of microorganisms and microbial fermentation, in particular to a protease produced by bacteria and having a low optimum enzyme reaction temperature, a microbial strain producing the enzyme and a method for producing the enzyme. Background technique [0002] Protease is currently the most widely used enzyme, accounting for 60-65% of the global enzyme industry market share. The proteases currently used are basically mesophilic enzymes. Compared with mesophilic protease, low temperature protease can maintain activity at low temperature (<10°C), K cat value and physiological efficiency (K cat / K m ) is high and has higher catalytic activity and other characteristics, so low-temperature protease has more application value than medium-temperature protease. Low-temperature protease is used in food, detergent, cosmetics, aquatic feed and other industries, and has the advantages that medium-temperature protease cannot r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/52C12R1/38
Inventor 陈吉刚杨季芳毛芝娟
Owner ZHEJIANG WANLI UNIV
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