One kind production alkaline pectate lyase genetic engineering bacterium and its structure and application

One kind production alkaline pectate lyase genetic engineering bacterium and its structure and application

  • CN 102,191,212 B
  • Filed: 03/29/2011
  • Issued: 07/14/2017
  • Est. Priority Date: 03/29/2011
  • Status: Active Grant
First Claim
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1. a kind of method of fermenting and producing transelminase, it is characterised in that use production alkaline pectate lyase engineering bacteria forStrain is produced, seed culture medium is inoculated in, 37 DEG C of cultivation temperature, shaking speed 200r/min cultivates 8h;

  • By the seed after cultureInoculation fermentation culture medium, condition of culture is 37 DEG C, 200r/min, as thalline OD600After reaching 0.8,1mM is added into culture mediumIPTG, 20mM calcium chloride, 0.1% glycine and 0.1%triton-100, continue the 2h that ferments;

    Or 5g/L lactose is only added, continueFerment 4h;

    It is described production alkaline pectate lyase engineering bacteria construction method be:

    (1) complete artificial synthesized aspergillus nidulans alkaline pectate lyase gene (pel), nucleotide sequence such as SEQ ID are utilizedShown in NO.1;

    (2) step (1) is obtained into pel sequences and is cloned into pET-20b (+), obtain recombinant expression plasmid pel/pET-20b (+);

    (3) recombinant expression plasmid pel/pET-20b (+) the Transformed E scherichia coli BL 21 for obtaining step (2)(DE3) recombination bacillus coli pel/pET-20b (+)/Escherichia coli BL 21 (DE3), are obtained, are named asEscherichia coli pel。

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