A kind of bacillus licheniformis gene shuffling strain and application of high-yield bacitracin a

A Bacillus licheniformis and gene shuffling technology, applied in the field of bioengineering, can solve problems such as uncontrollable excitation ion ratio, unclear gene mutation mechanism, etc., and achieve the effects of simple and accurate detection method, detection yield, and yield.

Active Publication Date: 2020-09-15
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

Although ARTP technology has been proved to be effective in causing gene mutations, the mechanism of gene mutations is still unclear, and the ratio of various excited ions produced cannot be controlled.

Method used

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  • A kind of bacillus licheniformis gene shuffling strain and application of high-yield bacitracin a
  • A kind of bacillus licheniformis gene shuffling strain and application of high-yield bacitracin a
  • A kind of bacillus licheniformis gene shuffling strain and application of high-yield bacitracin a

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Embodiment Construction

[0027] The present invention is described in detail below in conjunction with the thalline embodiment, and the following examples are used to illustrate the present invention, but are not used to limit the scope of the present invention.

[0028] 1. Bacitracin A high performance liquid phase detection method

[0029] HPLC conditions: use A: water + 0.1% trifluoroacetic acid; B: acetonitrile + 0.1% trifluoroacetic acid as the mobile phase for gradient elution, elution conditions: phase A linearly increases from 30% to 40%, and the time is 20 min. The injection volume was 20 μL, the detection wavelength was 220 nm, and the column temperature was 35 °C.

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Abstract

The invention discloses a bacillus licheniformis gene-recombined strain with high production of bacitracin A and its application. The invention adopts the plasma mutagenesis technology in the new breeding technology, combines with the traditional physical and chemical mutagenesis methods, and adopts the genome reorganization technology, so the effect is obvious and the yield is significantly improved. The gene-shuffled strain F2‑X1 for efficient fermentation and production of bacitracin A provided by the present invention reduces the cost of producing bacitracin by using Bacillus licheniformis as a fermentation strain. Moreover, the yield is significantly improved. After fermentation under the conditions, the yield of bacitracin A can reach 1.7 g / L, which is suitable for large-scale fermentation and production of bacitracin A. The HPLC detection method provided by the invention is simple and accurate, and is used for rapidly and accurately detecting the output of bacitracin A.

Description

technical field [0001] The invention relates to a high-yield bacitracin A gene-recombined bacterial strain of Bacillus licheniformis and its application, belonging to the technical field of bioengineering. Background technique [0002] Bacitracin is a cyclic peptide produced by Bacillus licheniformis or Bacillus subtilis that has a broad inhibitory effect on Gram-positive bacteria and some Gram-negative bacteria. According to the difference in amino acid composition and position, 15 homologues have been found. Among them, Bacitracin A, with the molecular formula C66H103N17O16S, is the main active substance and has the strongest biological activity. Bacitracin is widely used in feed agriculture due to the advantages of high efficiency, no residue, no toxic side effects, drug resistance and cross-resistance, and fast excretion. [0003] Due to the low vigor, harsh nutritional conditions and low production capacity of wild bacteria screened naturally, the production capacity o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12P21/04G01N33/68C12R1/10
CPCC12P21/02C12N1/205C12R2001/10G01N33/68
Inventor 陆兆新徐超吕凤霞别小妹赵海珍张充朱筱玉
Owner NANJING AGRICULTURAL UNIVERSITY
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