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Bacterial strain with self-flocculation capability as well as preparation method and application of bacterial strain

A technology of bacterial strains and bacteria, applied in the biological field, can solve the problems of restricting the large-scale application of PHA and high production costs, and achieve the effects of increasing oxygen transmission rate, promoting the growth of bacteria, and shortening the fermentation cycle

Pending Publication Date: 2022-06-28
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Claims
  • Application Information

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Problems solved by technology

As an environmentally friendly material, it will contribute to solving the energy and environmental crises brought about by the petroche

Method used

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  • Bacterial strain with self-flocculation capability as well as preparation method and application of bacterial strain
  • Bacterial strain with self-flocculation capability as well as preparation method and application of bacterial strain
  • Bacterial strain with self-flocculation capability as well as preparation method and application of bacterial strain

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Embodiment 1

[0067] Embodiment 1, the construction of ΔPS124 and its application in fermentation

[0068] 1. Construction of ΔPS124

[0069] Based on the two-plasmid knockout system of CRISPR-Cas9, the inventor has successively knocked out the three adjacent polysaccharide synthesis gene clusters PS1 (16861bp) and PS2 (22861bp) in Halomonas bluephagenesis TD1.0 through the gene large fragment knockout method. , PS4 (14851bp) for gene knockout ( Figure 5 In A), the ΔPS124 mutant was obtained. In the two-plasmid knockout system of CRISPR-Cas9 used, one plasmid is pQ08, and the other is a knockout plasmid pgVector that carries DNA and homology arms that transcribe sgRNA targeting the gene sequence to be knocked out.

[0070] Gene cluster PS1 (16861bp), whose sequence is sequence 1 in the sequence table, can encode IDs in NCBI as WP_039868450.1, WP_009721844.1, WP_009721843.1, WP_039868553.1, WP_109721841.1, WP_009721840.1, WP_009721839, respectively , WP_009721838.1, WP_009721837.1 protei...

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Abstract

The invention discloses a strain with self-flocculation capability and a preparation method and application thereof. The preparation method of the strain with the self-flocculation capability comprises the following steps: knocking out a polysaccharide synthesis gene in an original strain, or inhibiting the expression quantity of the polysaccharide synthesis gene in the original strain, or reducing the content or activity of protein coded by the polysaccharide synthesis gene in the original strain, so as to obtain a target recombinant strain. According to the invention, three polysaccharide synthesis gene clusters PS1, PS2 and PS4 are knocked out from halomonas to obtain a strain delta PS124 with self-flocculation capability, and the strain has an exopolysaccharide deletion phenotype and a self-flocculation phenomenon in a liquid culture medium.

Description

technical field [0001] The invention relates to a strain with self-flocculation ability and a preparation method and application thereof in the field of biotechnology. Background technique [0002] Polyhydroxyalkanoates (PHAs) are a class of polymer polyesters with diverse structures and properties synthesized by microorganisms. As an environmentally friendly material, it will contribute to solving the energy and environmental crisis brought about by the petrochemical economy, and has broad application prospects in human life, but the high production cost limits the large-scale application of PHA. Halomonas bluephagenesis screened from Xinjiang Salt Lake can accumulate a large amount of PHA particles PHB with 3HB as monomer in the cell, and has the characteristics of fast growth and high robustness, which is a major low-cost production of PHA. Breakthrough, the new technology for producing PHA by continuous fermentation without sterilization has been used in the factory pil...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/74C12P7/625C12R1/01
CPCC07K14/195C12N15/74C12P7/625
Inventor 韩静向华徐桐陈国强
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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