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Patent US10072255
Issued 2018-09-11
Microorganism Having Regulated Expression Of (r)-specific Enoyl-coa Hydratase Gene And Method For Producing Polyhydroxyalkanoate Copolymer Using Same
This invention relates to a microorganism that produces a polyhydroxyalkanoate (PHA) copolymer with a regulated monomer composition ratio and comprises a (R)-specific enoyl-CoA hydratase gene in the genome DNA, wherein a nucleotide sequence upstream of the (R)-specific enoyl-CoA hydratase gene comprises a modification consisting of a substitution(s), a deletion(s), an insertion(s), and/or an addition(s) of one or a plurality of nucleotides so that the expression of the (R)-specific enoyl-CoA hydratase gene is regulated, and to a method for producing a PHA copolymer using the microorganism.
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- 1. A method for producing a polyhydroxyalkanoate (PHA) copolymer having at least 5 mol % of 3-hydroxyhexanoate (3HH) monomer, the method comprising:
culturing a microorganism comprising a (R)-specific enoyl-CoA hydratase gene in the genome DNA, wherein a nucleotide sequence upstream of the (R)-specific enoyl-CoA hydratase gene in the genome DNA of the microorganism comprises a modification that comprises a substitution, a deletion, an insertion, and/or an addition of at least one nucleotide so that expression of the (R)-specific enoyl-CoA hydratase gene is regulated, and further comprising a polyhydroxyalkanoate synthase gene, wherein the modification comprises a modification of at least one nucleotide in a promoter sequence and/or a Shine-Dalgarno (SD) sequence originally existing upstream of the (R)-specific enoyl-CoA hydratase gene in the genome DNA; or an insertion or substitution of an expression regulatory DNA comprising a promoter sequence and/or SD sequence, at a site upstream of the (R)-specific enoyl-CoA hydratase gene in the genome DNA, so that the mol % of the 3HH monomer is higher than a control without the modification; wherein the modification comprises an insertion or substitution of an E. coli-derived promoter sequence selected from the group consisting of a trc promoter sequence, a lacUV5 promoter sequence, and a trp promoter sequence; and recovering a polyhydroxyalkanoate copolymer from the microorganism.
