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Serratia marcescens engineered strain with bvg90_08615 gene deletion

A technology of Serratia marcescens and engineering bacteria, which can be applied to microorganism-based methods, bacteria, peptides, etc., can solve the problems of low yield and hinder the industrialization process of microbial fermentation methods.

Active Publication Date: 2021-11-12
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there are still certain defects in the existing biological methods, among which the low yield is the most important defect hindering the industrialization of microbial fermentation methods

Method used

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  • Serratia marcescens engineered strain with bvg90_08615 gene deletion
  • Serratia marcescens engineered strain with bvg90_08615 gene deletion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Construction of Serratia marcescens engineering bacteria JNB5-1ΔBVG90_08615

[0031] Specific steps are as follows:

[0032] With the genome of Serratia marcescens JNB5-1 as a template, respectively 08615-D-U-F / 08615-D-U-R (SEQ ID NO:3 and SEQ ID NO:4) and 08615-D-D-F / 08615-D-D-R (SEQ ID NO:5 and SEQ ID NO: 6) are primers, obtain DNA fragment BVG90_08615-U (SEQ ID NO: 7) and DNA fragment BVG90_08615-D (SEQ ID NO: 8) by PCR amplification; synthetic aacC3 resistance gene (SEQ ID NO: 9); DNA fragment BVG90_08615-U, DNA fragment BVG90_08615-D and aacC3 resistance gene were sequentially connected by overlap extension PCR to obtain DNA fragment BVG90_08615-AacC3; DNA fragment BVG90_08615-AacC3 was digested with KpnI linearized pUTmini The plasmids were ligated after homologous recombination to obtain the recombinant plasmid pUTmini-BVG90_08615; the recombinant plasmid pUTmini-BVG90_08615 was transformed into Escherichia coli (Escherichiacoli) S17-1 to obtain the t...

Embodiment 2

[0036] Embodiment 2: the production of prodigiosin

[0037] Specific steps are as follows:

[0038] Taking Serratia marcescens JNB5-1 as a control, the single bacterium colonies of Serratia marcescens engineering bacteria JNB5-1ΔBVG90_08615, JNB5-1ΔBVG90_03840 and JNB5-1ΔBVG90_24040 obtained in Example 1 were picked and inoculated into LB liquid medium ( Contains 50μg·mL -1 Apramycin and 50 μg·mL -1 Clindamycin), cultured with shaking at 37°C and 180rpm until the early logarithmic growth phase (OD 600 =0.6), to obtain a seed solution; the seed solution was inoculated into LB liquid medium with a 4% (v / v) inoculation amount, and fermented for 24 hours at 30° C. and 180 rpm to obtain a fermentation broth.

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Abstract

The invention discloses Serratia marcescens engineering bacteria with BVG90_08615 gene deletion, and belongs to the field of biotechnology. The present invention provides a Serratia marcescens engineering bacterium JNB5‑1ΔBVG90_08615 that can produce prodigiosin at a high rate. obtained from the gene encoding the transcriptional regulator BVG90_08615, the Serratia marcescens engineered bacterium JNB5‑1ΔBVG90_08615 was inoculated into LB liquid medium and fermented for 24 hours, and the prodigiosin production in the fermentation broth could be as high as 58.68mg / L, 1.15 times that of wild-type Serratia marcescens JNB5‑1.

Description

technical field [0001] The invention relates to Serratia marcescens engineering bacteria with BVG90_08615 gene deletion, and belongs to the field of biotechnology. Background technique [0002] Prodigiosin (PG) is a class of secondary metabolites produced by microorganisms. Studies have found that prodigiosin has a variety of biological activities, including: anti-bacteria, anti-dysentery, anti-tumor and immunosuppression, etc. Therefore, prodigiosin has great application value in fields such as pharmaceutical development. [0003] At present, the methods for producing prodigiosin mainly include chemical synthesis and microbial fermentation. Among them, the chemical synthesis method mainly obtains prodigiosin through tandem conjugate addition and high-temperature dehydrogenation. However, due to complex and difficult pathways and low yields, chemical synthesis is difficult to achieve large-scale industrial production. The principle of microbial fermentation is mainly to o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12P17/16C12R1/43
CPCC07K14/24C12P17/165
Inventor 饶志明潘学玮杨套伟尤甲甲易敢峰付维来徐美娟张显邵明龙
Owner JIANGNAN UNIV