Recombined bacterial strain for modifying specificity of 1,3-propanediol redoxase coenzyme and application thereof

Recombined bacterial strain for modifying specificity of 1,3-propanediol redoxase coenzyme and application thereof

  • CN 101,643,739 B
  • Filed: 10/22/2008
  • Issued: 04/10/2013
  • Est. Priority Date: 10/22/2008
  • Status: Active Grant
First Claim
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1. the specific remodeling method of the coenzyme of a dhaT is characterized in that:

  • 1, the ammediol oxidoreductase gene is from Klebsiella, citric acid Pseudomonas or fusobacterium, sequence with SEQ ID NO;

    1, or the degeneracy sequence of SEQ ID NO;

    1, or coded sequence is because replacing, lack, insert and/or to add one or several amino-acid residue different with the coded sequence of SEQ ID NO;

    1, but still has the nucleotide sequence of the enzyme of same enzyme activity;

    1, ammediol oxydo-reductase mutant enzyme has the sequence of SEQ ID NO;

    2, perhaps because replacing, lack, insert and/or to add one or several amino-acid residue different with the sequence of SEQ ID NO;

    2, but still has the amino acid acid sequence of the enzyme of same enzyme activity;

    1, ammediol oxydo-reductase mutant enzyme gene has the sequence of SEQ ID NO;

    3, or the degeneracy sequence of SEQ ID NO;

    3, or coded sequence is because replacing, lack, insert and/or to add one or several amino-acid residue different with the coded sequence of SEQ ID NO;

    3, but still has the nucleotide sequence of the enzyme of same enzyme activityBut employed promotor is constitutive promoter or inducible promoter, constitutive promoter is selected from protein kinase promotor, fixed nitrogen promotor or Protosol promotor, but inducible promoter is selected from hybrid promoter or the trp promoter of Lac operon, phage promoter, lactose and tryptophane;

    The carrier framework of employed recombinant vectors is selected from pBR322, pUC18, pET series or pDK serial carrier;

    Employed host cell is selected from Klebsiella, citric acid Pseudomonas or fusobacterium;

    Fermentation mode adopts batch fermentation, fedbatch culture fermentation or continuously ferments;

    Contain carbon source, nitrogenous source, inorganic salt and VITAMIN in seed and the fermention medium;

    Fermentation is to pass into aerobic fermentation, the micro-aerobe fermentation that air carries out or pass into the anaerobically fermenting that nitrogen carries out in microbial cultivation process in microbial cultivation process;

    Fermentation inoculum size 1~

    12%, 20~

    50 ℃

    of culture temperature, air flow is 0.1~

    1.0vvm during fermentation, regulates pH and maintains 5.0~

    9.0, mixing speed is 80~

    350rpm.

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