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Method for preparing alkali protease from serratieae and special culture medium thereof

A technology for Serratia and Serratia marcescens, which is applied in the biological field and can solve the problems of low yield, inconvenient extraction, and not being widely used

Inactive Publication Date: 2011-05-25
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] Alkaline protease is widely used in enzyme-added detergents, leather, silk and other industries. At present, the main industrial enzyme-producing strains are Bacillus licheniformis and Bacillus pumilus, but spore-like strains will produce a large number of spores while producing enzymes , which brings inconvenience to the extraction of the enzyme, and the post-processing is also more cumbersome
Serratia marcescens is a Gram-negative non-spore bacterium that can produce alkaline protease, and this protease molecule is relatively large, easy to purify, and is an excellent candidate strain for industrial production of alkaline protease; in addition, Serratia The protease produced by Bacillus also has anti-tumor drug properties; but due to the low activity and low yield of the protease produced by it, it has not been widely used in industrial production

Method used

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  • Method for preparing alkali protease from serratieae and special culture medium thereof
  • Method for preparing alkali protease from serratieae and special culture medium thereof

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Experimental program
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Effect test

Embodiment 1

[0017] Embodiment 1: the detection of Serratia sp. (Serratia sp.) KMR-3 bacterial strain alkaline protease:

[0018] A Serratia sp. KMR-3 strain with high resistance to various heavy metal ions was isolated from the wastewater around the Kunming smelter. The Serratia sp. (Serratia sp.) KMR-3 bacterial strain has been preserved on August 26, 2008 in the "Common Microorganism Center of China Microbiological Culture Collection Management Committee" (Beijing, China), and the preservation number is: CGMCC No.2636 .

[0019] Detection of KMR-3 strain alkaline protease: apply 50 μl of freshly cultivated KMR-3 bacterial solution in 5 mL of LB liquid medium with different pH (1-14), culture at 25°C, 150 rpm for 24 hours, and observe the growth pH of the bacteria and protease production range. The protease production of KMR-3 strain under different pH conditions was determined by Folin's phenol method. The results show that: KMR-3 strain can grow in LB liquid medium in the range of p...

Embodiment 2

[0020] Embodiment 2: the optimization of medium composition:

[0021] Preliminary optimization of the improved LB liquid medium: LB liquid medium (tryptone 10g / L, yeast extract 5g / L, NaCl 10g / L) was used as the initial medium, the fermentation temperature was 15°C, and the pH value was 8.0. The content of the preliminary optimization experiment is: LB liquid medium, improved LB liquid medium without tryptone, improved LB liquid medium without yeast extract, improved LB liquid medium without NaCl, respectively at 0, 6, and 12 , 24, 36, 48, 60, 72, 84 and 96h, the fermentation broth was sampled, the supernatant was obtained by centrifugation, and then the OD was measured according to the protease activity assay method 660 The value is initially estimated protease production, and finally converted to protein concentration, the results are as follows figure 1 The results show that: LB medium lacking NaCl can promote the strain to produce enzyme in advance, and the amount of enzym...

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Abstract

The invention relates to a production method for preparing alkali protease from serratieae, which belongs to the technical field of biology. Serratia sp. KMR-3 strains are cultured for 24h at 25 DEG C and the pH value being 7 to 10 in an improved NaCl-free luria-bertani (LB) liquid culture medium with 10g / l tryptone, 2.5g / l yeast extracts and water as ingredients. The culture medium for preparing the alkali protease from the serratieae is the culture medium using the tryptone, the yeast extracts and the water as the ingredients. The culture medium has the characteristic of relatively little ingredient consumption, also has an effect of promoting the serratieae to fast produce the alkali protease, is applicable to the fields of germ secondary metabolites in modern biological engineering, laboratory culture and the like and can provide the study basis for the industrial production of the alkali protease.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for producing alkaline protease by Serratia, a culture medium specially used for the method, and the culture medium uses peptone and yeast extract as C and N sources and energy sources. Background technique [0002] Alkaline protease is widely used in enzyme-added detergents, tanning, silk and other industries. At present, the main industrial enzyme-producing strains used are Bacillus licheniformis and Bacillus pumilus, but spore-like strains will produce a large number of spores while producing enzymes , which brings inconvenience to the extraction of the enzyme, and the post-processing is also more cumbersome. Serratia marcescens is a Gram-negative non-spore bacterium that can produce alkaline protease, and this protease molecule is relatively large, easy to purify, and is an excellent candidate strain for industrial production of alkaline protease; in addition, Serratia m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/52C12N1/20C12R1/43
Inventor 魏云林杨琳琳季秀玲林连兵
Owner KUNMING UNIV OF SCI & TECH