Ferulic acid producing engineering strain, construction method and biotransformation method
A production engineering and construction method technology, applied in the field of biotransformation, catalyzing eugenol to produce ferulic acid engineering strains, can solve the problems of high energy consumption, high price, a large amount of acid-base waste, etc., and achieve simple implementation methods and high industrial efficiency the effect of value
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[0039] A method for constructing ferulic acid production engineering strains, comprising the following steps:
[0040] 1. Construction of strains containing recombinant plasmid pET24a-vaoA
[0041] 1) Plasmid construction
[0042] Plasmid pET24a(+): a product of Novagen, with a size of 5.31kb, containing a kanamycin resistance gene, a lactose repressor lac I gene, a T7 promoter, and multiple restriction endonuclease sites.
[0043] The DNA fragment of the vanillyl oxidase gene (vaoA, 1683bp) was digested with Nde I and Bam HI. The vector pET24a(+) was double digested with Nde I and Bam HI. Enzyme digestion system: DNA 43 μL, buffer R 5 μL, Bam HI 1 μL, Nde I 1 μL, incubated at 37°C for 3 hours.
[0044] The digested DNA fragments were gel-recovered, and the vaoA and carrier DNA fragments were ligated with T4 ligase. The ligation system was as follows: 7.5 μL of vaoA, 1.5 μL of pET24a vector, 1 μL of buffer, and 1 μL of T4 ligase. Transformed into Escherichia coli host stra...
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