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
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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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