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Method for improving acid stress resistance of Serratia marcescens

A technology of Serratia marcescens and acid stress, which is applied in the field of improving the ability of Serratia marcescens to resist acid stress, and can solve by-product accumulation, osmotic pressure stress, and influence on the growth and metabolism of Serratia marcescens, etc. problem, to achieve good effect and good acid stress resistance

Active Publication Date: 2020-02-28
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the addition of alkaline substances often leads to the accumulation of by-products during the fermentation of Serratia marcescens, and the salts formed by these by-products will again cause the cells to be in a hypertonic environment, resulting in osmotic stress The production of Serratia marcescens affects the growth and metabolism of Serratia marcescens again

Method used

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  • Method for improving acid stress resistance of Serratia marcescens
  • Method for improving acid stress resistance of Serratia marcescens
  • Method for improving acid stress resistance of Serratia marcescens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1: Construction of recombinant bacterial strain

[0037] Specific steps are as follows:

[0038] (1) chemically synthesize the xrpA gene whose nucleotide sequence is shown in SEQ ID NO.2 (xrpA gene is the gene encoding the DNA-binding protein XrpA, and the DNA-binding protein XrpA is a protein used to recognize and bind DNA on the microbial cell membrane, belonging to XRE family), the xrpB gene whose nucleotide sequence is shown in SEQ ID NO.3 (xrpB gene is the gene encoding XrpB, XrpB is a protein used to recognize and bind DNA on the microbial cell membrane, and belongs to the XRE family), nucleotide The sequence is as shown in the xrpC gene of SEQ ID NO.4 (the xrpC gene is the gene encoding XrpC, and XrpC is a protein used to recognize and bind DNA on the microbial cell membrane, belonging to the XRE family), and the nucleotide sequence is as in SEQ ID NO.5 The xrpD gene shown (the xrpD gene is a gene encoding XrpD, and XrpD is a protein used to recognize...

Embodiment 2

[0042] Embodiment 2: the growth performance test of recombinant bacterial strain

[0043] Specific steps are as follows:

[0044] Only the recombinant Serratia marcescens / pET-28a-Vector (control) containing blank plasmid pET-28a (+) and the recombinant Serratia marcescens / pET-28a-Vector (control) obtained in Example 1 were xrpA, recombinant Serratia marcescens / pET-28a-xrpB, recombinant Serratia marcescenss / pET-28a-xrpC, recombinant Serratia marcescens / pET-28a-xrpD, respectively Streak on LB solid medium and culture at 30°C for 16h; pick positive transformants and inoculate them in LB liquid medium containing 50μg / mL kanamycin, culture at 30°C and 180rpm for 12h to obtain seeds solution; the seed solution was inoculated in LB liquid medium to control the initial OD 600 =0.1, cultured at 30°C and 180rpm, took samples from the culture solution every two hours to measure the OD of the culture solution 600 value, draw the growth curve (see figure 2 ).

[0045] The result is a...

Embodiment 3

[0046] Example 3: Tolerance test of recombinant strains under acid stress conditions

[0047] Specific steps are as follows:

[0048] Only the recombinant Serratia marcescens / pET-28a-Vector (control) containing blank plasmid pET-28a (+) and the recombinant Serratia marcescens / pET-28a-Vector (control) obtained in Example 1 were xrpA and recombinant Serratia marcescens / pET-28a-xrpC were respectively streaked on LB solid medium, cultured at 30°C for 16 hours, and single colonies were obtained; single colonies were picked and inoculated in LB liquid medium respectively, Cultivate under the conditions of 30°C and 180rpm for 12h to obtain seed liquid; inoculate the seed liquid into LB liquid medium according to the inoculation amount of 1% (v / v), and cultivate under the conditions of 30°C and 180rpm for 12h to obtain Culture fluid: collect part of the culture fluid, centrifuge to collect the cells, wash the collected cells twice with 0.85% saline, and then resuspend them in LB liqu...

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Abstract

The invention discloses a method for improving acid stress resistance of Serratia marcescens, and belongs to the technical field of gene engineering and microbial engineering. DNA (deoxyribonucleic acid) binding proteins XrpA are over-expressed in the Serratia marcescens to improve the acid stress resistance of the Serratia marcescens. The recombinant Serratia marcescens prepared by the method iscultured in an environment with the pH (potential of hydrogen) of 4.0 for 12 hours, and then the survival rate of the recombinant Serratia marcescens is 1.4 times that of wild Serratia marcescens.

Description

technical field [0001] The invention relates to a method for improving the acid stress resistance of Serratia marcescens, which belongs to the technical fields of genetic engineering and microbial engineering. Background technique [0002] Serratia marcescens is an important industrial strain, which is often used to produce prodigiosin and 2,3-butanediol, which are widely used in the preparation of food, medicine and daily chemical products. Fermented products have broad market prospects. [0003] However, in the fermentative production process of Serratia marcescens, it is inevitable to face a variety of environmental stresses from the external environment such as acid stress, ethanol stress, oxygen stress, salt stress, etc. These environmental stresses Both seriously affect the physiological activity of Serratia marcescens (Serratia marcescens), and then seriously limit the fermentation performance of Serratia marcescens (Serratia marcescens). [0004] Among the many env...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/74C12N1/21C12P17/16C12P7/18C12R1/43
CPCC12N15/74C12P7/18C12P17/165
Inventor 饶志明孙长皓潘学玮杨套伟徐美娟张显邵明龙
Owner JIANGNAN UNIV
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