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Patent US10233468
Issued 2019-03-19
High Molecular Weight Pha-producing Microbe And Method Of Producing High Molecular Weight Pha Using Same
An object of the present invention is to provide a microorganism strain that accumulates a high molecular weight PHA, and a PHA production method using the microorganism. The present invention provides a method for producing a PHA copolymer, which includes culturing a microorganism, wherein at least a portion of either of the following genes (a) and (b) of the microorganism has been altered by substitution, deletion, insertion, and/or addition to reduce or eliminate the activity of a PHA degrading enzyme encoded by the gene: (a) a PHA degrading enzyme gene encoding the amino acid sequence of SEQ ID NO:2 in the sequence listing; and (b) a gene encoding a polypeptide having at least 85% sequence identity to the amino acid sequence of SEQ ID NO:2 in the sequence listing and having PHA degrading enzyme activity.
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- 1. A method for producing a polyhydroxyalkanoate (PHA) copolymer having an increased weight average molecular weight, comprising:
(1) culturing a PHA producing microorganism, wherein at least a portion of a gene encoding a PHA degrading enzyme of the microorganism has been altered by at least one of substitution, deletion, insertion, and addition, to reduce or eliminate an activity of the PHA degrading enzyme, wherein the PHA degrading enzyme has (a) the amino acid sequence of SEQ ID NO:2, or (b) an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:2 and has a PHA degrading enzyme activity, wherein the microorganism belongs to the genus Cupriavidus, and (2) producing the PHA copolymer, wherein the weight average molecular weight of the produced PHA copolymer is increased compared to the weight average molecular weight of the produced PHA copolymer in a microorganism wherein the gene encoding the PHA degrading enzyme has not been altered, and wherein the produced PHA copolymer comprises at least two different type of units selected from the group consisting of 3-hydroxyhexanoic acid (3HH), 3-hydroxybutyric acid (3HB), 3-hydroxyvaleric acid (3HV), 3-hydroxyoctanoic acid (3HO), 4-hydroxybutyric acid (4HB), and a combination thereof.
- 16. A method for producing a polyhydroxyalkanoate (PHA) copolymer having an increased weight average molecular weight, the method comprising:
(1) culturing a PHA producing microorganism, wherein the microorganism belongs to the genus Cupriavidus and in which a PHA synthase comprising the amino acid sequence of SEQ ID NO:15 has been introduced, wherein (a) a phaZ6 gene and a phaZ1 gene on the chromosome of Cupriavidus have been deleted; (b) a phaZ6 gene and a phaZ2 gene on the chromosome of Cupriavidus have been deleted; or (c) a phaZ6 gene, a phaZ1 gene, and a phaZ2 gene on the chromosome of Cupriavidus have been deleted; and (2) producing the PHA copolymer, wherein the produced PHA copolymer comprises at least two different type of units selected from the group consisting of 3-hydroxyhexanoic acid (3HH), 3-hydroxybutyric acid (3HB), 3-hydroxyvaleric acid (3HV), 3-hydroxyoctanoic acid (3HO), 4-hydroxybutyric acid (4HB), and a combination thereof; and wherein the weight average molecular weight of the produced PHA copolymer is increased compared to the weight average molecular weight of the produced PHA copolymer in a Cupriavidus microorganism wherein the gene encoding the PHA degrading enzyme has not been altered.
