Green alga lipid-accumulating variant and use of the same

Patent metadata

Field	Value
Publication number	US-10087496-B2
Application number	US-201415037026-A
Country	US
Kind code	B2
Filing date	Nov 7, 2014
Priority date	Nov 19, 2013
Publication date	Oct 2, 2018
Grant date	Oct 2, 2018

Abstract

A green alga variant having a dual-specificity tyrosine-phosphorylation regulated protein kinase activity that is reduced compared to a dual-specificity tyrosine-phosphorylation regulated protein kinase activity of a parental strain is provided. The green alga variant increases a total amount of a lipid production per unit time and per unit culture area compared to a total amount of a lipid production of the parental strain. A dual-specificity tyrosine-phosphorylation regulated protein kinase of the parental strain is a protein having an amino acid sequence with at least 50% sequence identity with the amino acid sequence of an active site and a substrate recognition site of SEQ ID NO: 4 and having the dual-specificity tyrosine-phosphorylation regulated protein kinase activity.

First claim

What is claimed is:

1. A green alga variant comprising:
a genetic variant of a gene encoding a dual-specificity tyrosine-phosphorylation regulated protein kinase (DYRK), an activity of the DYRK being lost or reduced compared to an activity of DYRK activity of a parental strain,
wherein:
the green alga variant belongs to Pseudochoricystis ellipsoidea;
the green algae variant has an increase of a total amount of a lipid production per unit time and per unit culture area compared to a total amount of a lipid production of the parental strain;
a dual-specificity tyrosine-phosphorylation regulated protein kinase of the parental strain is a protein having an amino acid sequence with at least 50% sequence identity with the amino acid sequence of an active site and a substrate recognition site of SEQ ID NO: 4, and having the dual-specificity tyrosine-phosphorylation regulated protein kinase activity; and
the green alga variant is generated by:
contacting genomic DNA encoding for a DYRK in the parental strain with a mutagenic substance to generate a plurality of genetic variants;
screening the plurality of genetic variants for genetic variants having increased lipid content; and
isolating the green alga variant having a genetic variant of the gene encoding the DYRK from the genetic variants having increased lipid content.

2. The green alga variant according to claim 1 , which is a green alga in which the gene encoding a dual-specificity tyrosine-phosphorylation regulated protein kinase is disrupted.

3. A lipid production method, comprising:
culturing the green alga variant according to claim 1 .

4. The green alga variant according to claim 1 , wherein:
the gene encoding the dual-specificity tyrosine-phosphorylation regulated protein kinase has either a nucleotide substitution mutation or a nucleotide deletion mutation as the genetic variant.

5. The green alga variant according to claim 1 , wherein the genetic variant of the gene encoding the DYRK comprises a base substitution in an intron that results in a splice variant, a base substitution in an exon that results in an amino acid mutation, or a deletion in an exon that results in a frameshift mutation.

Assignees

• Denso Corp
• Univ Chuo

Inventors

• Hayakawa Jumpei
• Ide Yoko
• Harayama Shigeaki
• Yasui Hidehiko

Classifications

• C12P7/6463

  obtained from glyceride producing microorganisms, e.g. single cell oil · CPC title

• C12N15/01

  Preparation of mutants without inserting foreign genetic material therein; Screening processes therefor · CPC title

• Y02E50/13

  Cross-Sectional Technologies · mapped topic

• C12N1/12 Primary

  Unicellular algae; Culture media therefor (as new plants A01H13/00) · CPC title

• C12N9/1205

  Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases · CPC title