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A high-efficiency enzyme-producing Bacillus subtilis strain lc1-1 and its enzyme-producing method and application

A technology of Bacillus subtilis and chitosanase, applied in the biological field, can solve problems such as poor enzyme production effect, and achieve the effects of optimizing process process, improving enzyme production efficiency, and stable and efficient enzyme production rate.

Active Publication Date: 2022-07-15
MARINE BIOMEDICAL RES INST OF QINGDAO CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main disadvantage of screening enzyme-producing microorganisms from nature is that the enzyme-producing effect is poor, and some enzyme-producing strains need to be induced to achieve high enzyme-producing effects

Method used

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  • A high-efficiency enzyme-producing Bacillus subtilis strain lc1-1 and its enzyme-producing method and application
  • A high-efficiency enzyme-producing Bacillus subtilis strain lc1-1 and its enzyme-producing method and application
  • A high-efficiency enzyme-producing Bacillus subtilis strain lc1-1 and its enzyme-producing method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1 Bacillus subtilis LC1-1 strain identification

[0066] Pick a single colony of LC1-1 and inoculate it on the LB solid plate, and cultivate it on the LB solid medium for 24 hours. The colony shape of LC1-1 in the LB medium is as follows: figure 1 As shown in (a), the colonies are irregular in shape, with dark and opaque surfaces; they can be wrinkled, and under the electron microscope, the bacteria are straight rods. The physiological and biochemical characteristics of LC1-1 refer to "Bergey's OF SystematicBacteriology Second Edition" (Ninth Edition), and the experimental strains were subjected to Gram staining, spore staining, strain motility observation, catalase test, ethanol methyl methanol (V.P. ) test, nitrate reduction test, litmus milk test, anaerobic growth test and other aspects of physiological and biochemical experiments, the results show that after the spore staining test, LC1-1 shows spore mesogenesis, and the rest are positive results.

[0067] ...

Embodiment 2

[0074] Example 2 Chitosanase-producing performance of Bacillus subtilis LC1-1 on a plate at 65°C

[0075] The primary screening medium uses chitosan as the only carbon source, and a degraded transparent circle will be formed after the chitosan is degraded. The production of transparent circles in the primary screening medium is as follows figure 1 (b), in figure 1 (b) The ratio of the diameter of the transparent circle to the diameter of the colony is 3.

Embodiment 3

[0076] Example 3 Optimization of fermentation production conditions for chitosanase

[0077] (1) Single factor experiment

[0078] Single factor experiments were carried out on factors such as medium components (carbon source, nitrogen source, optimum carbon source concentration, optimum nitrogen source concentration) and fermentation conditions (temperature, pH, rotational speed, liquid volume, inoculum volume) (see the results in figure 2).

[0079] From Figure 2(a)-(i), it can be obtained that the optimum inoculum size is 2%, the optimum fermentation temperature is 35°C, the optimum initial fermentation pH is 5.5, the optimum rotation speed is 180rpm, the liquid loading amount is 20%, and the carbon The source concentration was 20g / L, and the nitrogen source concentration was 15g / L. Based on this, Minnimum-Run Equireplicated Res IV design and response surface optimization design were carried out.

[0080] (2) Minnimum-Run Equireplicated Res IV design

[0081] Design-Expe...

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Abstract

The present invention relates to a high-efficiency chitosanase-producing Bacillus subtilis strain LC1-1 and an enzyme-producing method and application thereof. The strain was deposited in the General Microorganism Center of the China Committee for the Preservation of Microorganisms on February 4, 2021, with a deposit number of It is CGMCC No.21809. The Bacillus subtilis strain LC1-1 can produce chitosanase at a temperature of 25-65°C and a pH of 4.5-8. The enzyme production process does not require chitosan induction, and is optimized by carbon and nitrogen sources. By optimizing the composition of the medium, the obtained chitosanase has high yield and good stability, and can degrade chitosan to generate monosaccharide, chitotriose and chitotetraose, and has a good application prospect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a high-yielding chitosanase-producing Bacillus subtilis LC1-1 and an enzyme-producing method and application thereof. Background technique [0002] Chitosan refers to a chitin product with a degree of deacetylation higher than 55%, and is the only natural cationic polysaccharide found so far. Although many studies have shown that chitosan has very important applications in biomedicine, agriculture, wastewater and waste treatment, diet (food preservatives and preservatives, etc.), and cosmetics, due to its large molecular weight and poor water solubility, its Applications are also limited. Chitosan oligosaccharide, also known as oligomeric chitosan, is the product of further degradation of chitosan. The molecular weight is generally lower than 3000Da, the degree of polymerization is between 2 and 20, the water solubility is good, and the viscosity is low. Glycans. In addition to th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/42C12R1/125
CPCC12N9/2434C12Y302/01132
Inventor 何增国郭若琳汤伟孙晓雯张军唐涛
Owner MARINE BIOMEDICAL RES INST OF QINGDAO CO LTD