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High-temperature and strong base resistant clausii bacillus cereus, excretive proteolytic enzyme and preparation

A technology of Bacillus clausii and Bacillus, applied in the direction of bacteria, hydrolytic enzymes, and microbial-based methods, can solve the problem of low anti-oxidation and anti-SDS performance, enzymatic properties that are difficult to meet the needs of the development of the washing industry, and unsatisfactory It has large-scale industrial development potential and other issues, and achieves the effects of good thermal stability and pH stability, good thermal stability, and wide pH stability

Inactive Publication Date: 2008-09-24
XINJIANG UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Preliminary domestic studies have also reported strains that can secrete proteases that are stable to SDS and oxidants, but the performance of anti-oxidation and anti-SDS is not high, and the main enzymatic properties of domestic alkaline enzymes are difficult to meet the needs of the development of the washing industry. potential for industrial development, the source of large-scale industrial alkaline enzymes mainly depends on foreign imports

Method used

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  • High-temperature and strong base resistant clausii bacillus cereus, excretive proteolytic enzyme and preparation
  • High-temperature and strong base resistant clausii bacillus cereus, excretive proteolytic enzyme and preparation
  • High-temperature and strong base resistant clausii bacillus cereus, excretive proteolytic enzyme and preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0132] Example 1: Cultivation and screening of high-temperature alkaline protease-producing bacteria B.ciausii XJU-5

[0133] 1. Sample collection

[0134] Samples were collected from saline water in Turpan, Xinjiang.

[0135] 2. Isolation of strains

[0136] Prepare plate medium (g / l): 2.0% wheat straw, 0.2% NH 4 NO 3 , 0.2%K 2 HPO 4 , 0.02% MgSO 4 ·7H 2 O, 0.5% yeast extract, 1.8% agar powder, pH10.5. The above medium was added at 1.05kg / cm 2 , 121°C, autoclaved for 20 minutes.

[0137] Centrifuge the collected water sample at 8000rpm for 10min, discard the supernatant, dissolve the precipitate with sterilized physiological saline, and prepare the suspension at 10 1 , 10 3 , 10 4 , 10 5 , 10 7 Make 5 dilution gradients, take 0.2mL of each dilution sample solution and spread evenly on the above medium (pH10.5) plate, culture at 38°C for 18 hours to obtain a single colony, pick a single colony with an inoculation needle Draw a line and connect to the above-menti...

Embodiment 2

[0142] Embodiment 2: Fermentation culture of high temperature alkaline protease producing bacteria B.clausii XJU-5

[0143] The culture medium is the basis of fermentation technology. On the basis of laboratory research, the medium screening and optimization of industrial pilot test research are carried out, and the optimization is carried out on the basis of basic fermentation culture medium.

[0144] 2. Basic fermentation medium: peptone 10g, yeast extract powder 5g, sodium chloride 5g, dipotassium hydrogen phosphate 1g, magnesium sulfate crystal 0.2g and water 1000ml.

[0145] Note: pH10.0-11.0, adjust the pH of the sterilized medium with sterilized 10% sodium carbonate solution.

[0146] (1) Optimization of fermentation medium for enzyme production

[0147] 1. Effect of carbon source on enzyme production

[0148] Choose no carbon source, glucose, yeast extract, corn flour, sucrose, and starch to replace the yeast extract powder in the basic culture solution to make diffe...

Embodiment 3

[0159] Embodiment 3: the measurement method of high temperature alkaline protease activity

[0160] According to the "Industrial Standards of the People's Republic of China" QB / T 1803, 1804-93, QB / T 1805, 1806-93 issued by the Ministry of Light Industry of the People's Republic of China on 1993-07-29 and implemented on 1994-03-01, industrial enzymes General test method for preparations, during the test, the temperature is set at 40°C and the pH is 10.5.

[0161] 1.1 Definition

[0162] 1 g of solid enzyme powder (or 1 mL of liquid enzyme), under certain temperature and pH conditions, hydrolyzes casein for 1 min to produce 1 μg of tyrosine, which is an enzyme activity unit, expressed in u / g (u / mL).

[0163] 1.2 Forint method

[0164] 3.2.1 Principle

[0165] Under certain temperature and pH conditions, protease hydrolyzes the casein substrate to produce amino acids containing phenolic groups (such as: tyrosine, tryptophan, etc.), and under alkaline conditions, the Folin reag...

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Abstract

The invention discloses high-temperature resistant and alkali resistant bacillus clausii CGMCC No. 1768, a production method thereof, and a method for producing high-temperature alkali protease and other enzyme preparations with the bacterial strain. The bacillus clausii has excellent tolerance, and can grow under extremely adverse environmental conditions as well as in the environment of NaCl of 0 to 20 percent, PH value from 5 to 14, and 4 DEG C to 85 DEG C. The bacillus clausii can survive in wide scope. By adopting the tolerance of the bacterial strain to extremely adverse environment, to research and develop the metabolites of the bacterial strain is very significant. Industrial production requirement can be met; the quality of enzyme preparations can be improved and stabilized; environmental pollution can be reduced; industry can be promoted to be stabilized and developed quickly by developing high temperature resistant and alkali resistant enzymes with excellent thermostability.

Description

field of invention [0001] The invention relates to the field of microbial fermentation and its product enzymes. Specifically, the present invention relates to a high-temperature and salt-resistant Bacillus clausii (Bacillus clausii), a secreted protease and a production and preparation method thereof. Background technique [0002] Alkaline protease has great application potential in industry, such as detergent, leather manufacturing, food processing, pharmaceuticals, and waste treatment industries. Alkaline protease can significantly improve product quality, save costs, and eliminate tanning, etc., by replacing traditional chemical reagents. Industrial pollutants, reduce environmental pollution, and play a pivotal role in industrial applications. [0003] In order to obtain alkaline protease with industrial value, in-depth research has recently been carried out on alkali-resistant strains derived from extreme environments. After research, alkalis with good thermal stability...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/54C12R1/07
Inventor 艾尔肯·热合曼邓爱华王宁易霞
Owner XINJIANG UNIVERSITY
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