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An engineering strain of Serratia marcescens and its application in the production of prodigiosin

A technology for Serratia marcescens and prodigiosin production, which is applied in the biological field and can solve the problems of low yield, hindering the industrialization process of microbial fermentation method and the like

Active Publication Date: 2022-05-24
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
For example, Kim et al. produced prodigiosin by inoculating Hahella chejuensis KCTC 2396 into the 2216 medium of marine bacteria broth for fermentation. However, using this method to ferment for 40 hours, only the concentration of prodigiosin in the fermentation broth Yield up to 28mg / L (specific references: Kim S J, Lee H K, Lee YK, et al. Mutant selection of Hahella chejuensis KCTC 2396 and statistical optimization of medium components for prodigiosin yield-up [J]. ):183-188.); Martha Ingrid Gutiérrez-Román et al fermented Serratia marcescens CFFSUR-B4 by inoculating it into a peanut-based medium to produce prodigiosin, however, using this method for 24h of fermentation, only the fermentation broth The production of prodigiosin and chitinases by tropical Serratia marcescens strains with potential to control plant pathogens [J].World Journal of Microbiology and Biotechnology,2012,28(1):p.145-153.)

Method used

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  • An engineering strain of Serratia marcescens and its application in the production of prodigiosin
  • An engineering strain of Serratia marcescens and its application in the production of prodigiosin
  • An engineering strain of Serratia marcescens and its application in the production of prodigiosin

Examples

Experimental program
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Effect test

Embodiment 1

[0035]Example 1: Construction of Serratia marcescens engineering strain JNB5-1ΔcpxR

[0036] Specific steps are as follows:

[0037] (1) Functional identification of the response regulatory protein CpxR

[0038] Using the genome of Serratia marcescens JNB5-1 as a template, the gene cpxR encoding the response regulatory protein CpxR was amplified by PCR (the amino acid sequence of the response regulatory protein CpxR is shown in SEQ ID No. The nucleotide sequence of the gene cpxR is shown in SEQ ID No. 2); the obtained gene cpxR encoding the response regulatory protein CpxR is connected with the pET-28a plasmid after double digestion by BamHI / Sac I to obtain a ligated product; The product was transformed into Escherichia coli BL21 to obtain the transformed product; the transformed product was spread on LB solid medium (containing 50 μg·mL -1 kanamycin), cultured upside down in a constant temperature incubator at 37 °C for 8 to 12 h to obtain transformants; pick the transforma...

Embodiment 2

[0056] Example 2: Production of prodigiosin

[0057] Specific steps are as follows:

[0058] With Serratia marcescens JNB5-1 as a control, a single colony of the Serratia marcescens engineering strain JNB5-1ΔcpxR obtained in Example 1 was picked and inoculated into LB liquid medium (containing 50 μg·mL). -1 Apramycin and 50 μg mL -1 Clindamycin), shake cultured at 37°C and 180rpm for 12h to obtain seed liquid; inoculate the seed liquid with 6% (v / v) inoculum into the fermentation medium, and at 30°C and 180rpm conditions Fermentation was carried out for 96h to obtain fermentation broth.

[0059] During the fermentation process, the content of prodigiosin in the fermentation broth was detected at intervals of 12h (see Figure 5 ), the results showed that the yield of prodigiosin in the fermentation broth obtained by the fermentation of Serratia marcescens JNB5-1 was 4.14 g / L at 96 h, and the yield of prodigiosin obtained by the fermentation of Serratia marcescens engineering...

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Abstract

The invention discloses a Serratia marcescens engineering bacterium and its application in the production of prodigiosin, belonging to the field of biotechnology. The invention provides a Serratia marcescens engineering bacterium JNB5‑1ΔcpxR capable of high prodigiosin production, and the Serratia marcescens engineering bacterium JNB5‑1ΔcpxR is obtained by knocking out Serratia marcescens JNB5‑1 Inoculated with the gene encoding the response regulator protein CpxR, the Serratia marcescens engineered bacterium JNB5-1ΔcpxR was inoculated into the fermentation medium and fermented for 96 hours, and the prodigiosin production in the fermentation broth could be as high as 5.83g / L , 41.9% higher than that of wild-type Serratia marcescens JNB5‑1.

Description

technical field [0001] The invention relates to a Serratia marcescens engineering bacterium and its application in producing prodigiosin, and belongs to the field of biotechnology. Background technique [0002] Prodigiosin is a kind of natural red pigment containing a tripyrrole skeleton structure, which is mainly produced by some microorganisms and has various biological activities such as anti-cancer, immunosuppression, and anti-insect. extensive attention of researchers. [0003] At present, the methods for producing prodigiosin mainly include chemical synthesis method and microbial fermentation method. Among them, the chemical synthesis method mainly obtains prodigiosin through tandem conjugate addition and high temperature dehydrogenation. However, due to the complex and difficult pathways and low yields, chemical synthesis is difficult to achieve large-scale industrial production. The principle of microbial fermentation is to obtain prodigiosin through microbial fer...

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 杨套伟饶志明孙杨徐美娟张显邵明龙付维来易敢峰
Owner JIANGNAN UNIV
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