US 12,305,199 B2Granted

Recombinant of Hydrogenophilus bacterium producing lactic acid

AssigneeUtilization of Carbon Dioxide Institute Co., Ltd.

InventorsHideaki Yukawa, Naoto Ohtani

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Abstract

When lactate dehydrogenase gene and/or malate/lactate dehydrogenase gene is/are introduced into a Hydrogenophilus bacterium as well as one or more of the three lactic acid-utilizing enzyme genes on the genome of the Hydrogenophilus bacterium is/are disrupted, lactic acid-producing ability is remarkably increased. The inventors of the present invention have identified the three lactic acid-utilizing enzyme genes of the Hydrogenophilus bacterium. When lactate permease gene is further introduced into the recombinant, lactic acid-producing ability is further increased. The recombinant of the present invention effectively produces lactic acid using carbon dioxide as a sole carbon source, and therefore, it is able to efficiently produce the material of biodegradable plastics, while solving global warming caused by increased emissions of carbon dioxide.

Claims17

Dependent claims

A recombinant Hydrogenophilus bacterium, which has a lactate dehydrogenase gene introduced thereinto, and in which one or more of three lactic acid-utilizing enzyme genes (a), (b), and (c) on a genome are disrupted so that lactic acid production in the bacterium is increased,

wherein the lactic acid-utilizing enzyme gene (a) comprises at least one selected from the group consisting of:
DNA comprising a nucleotide sequence having 90% or more identity to SEQ ID NO:
1;
DNA encoding a polypeptide comprising an amino acid sequence having 90% or more identity to SEQ ID NO:
2;
and
DNA encoding a polypeptide comprising an amino acid sequence having deletion, substitution, or addition of one to five of amino acids in the amino acid sequence of SEQ ID NO:
2;
wherein the lactic acid-utilizing enzyme gene (b) comprises at least one selected from the group consisting of:
DNA comprising a nucleotide sequence having 90% or more identity to SEQ ID NO:
3,
DNA encoding a polypeptide comprising an amino acid sequence having 90% or more identity to SEQ ID NO:
4;
and
DNA encoding a polypeptide comprising an amino acid sequence having deletion, substitution, or addition of 1 to 5 of amino acids in the amino acid sequence of SEQ ID NO:
4;
and
wherein the lactic acid-utilizing enzyme gene (c) comprises at least one selected from the group consisting of:
DNA comprising a nucleotide sequence having 90% or more identity to SEQ ID NO:
5;
DNA encoding a polypeptide comprising an amino acid sequence having 90% or more identity to SEQ ID NO:
6;
and
DNA encoding a polypeptide comprising an amino acid sequence having deletion, substitution, or addition of 1 to 5 of amino acids in the amino acid sequence of SEQ ID NO:
6.

22 more elements

A recombinant Hydrogenophilus bacterium, which has a malate/lactate dehydrogenase gene introduced thereinto, and in which one or more of three lactic acid-utilizing enzyme genes (a), (b), and (c) on a genome are disrupted so that lactic acid production in the bacterium is increased,

wherein the lactic acid-utilizing enzyme gene (a) comprises at least one selected from the group consisting of:
DNA comprising a nucleotide sequence having 90% or more identity to SEQ ID NO:
1,
DNA encoding a polypeptide comprising an amino acid sequence having 90% or more identity to SEQ ID NO:
2, and
DNA encoding a polypeptide comprising an amino acid sequence having deletion, substitution, or addition of one to five of amino acids in the amino acid sequence of SEQ ID NO:
2,
wherein lactic acid-utilizing enzyme gene (b) comprises at least one selected from the group consisting of:
DNA comprising a nucleotide sequence having 90% or more identity to SEQ ID NO:
3,
DNA encoding a polypeptide comprising an amino acid sequence having 90% or more identity to SEQ ID NO:
4, and
DNA encoding a polypeptide comprising an amino acid sequence having deletion, substitution, or addition of one to five of amino acids in the amino acid sequence of SEQ ID NO:
4,
wherein lactic acid-utilizing enzyme gene (c) comprises at least one selected from the group consisting of:
DNA comprising a nucleotide sequence having 90% or more identity to SEQ ID NO:
5,
DNA encoding a polypeptide comprising an amino acid sequence having 90% or more identity to SEQ ID NO:
6, and
DNA encoding a polypeptide comprising an amino acid sequence having deletion, substitution, or addition of one to five of amino acids in the amino acid sequence of SEQ ID NO:
6.

18 more elements

Dependencies are inferred from the claim text.