US 11,788,109 B2
Microorganism and method for producing gamma-glutamyl-valyl-glycine
Ayako Sasahara, Kawasaki (JP); Takayuki Ito, Kawasaki (JP); and Hiroyuki Nozaki, Kawasaki (JP)
Assigned to AJINOMOTO CO., INC., Tokyo (JP)
Filed by AJINOMOTO CO., INC., Tokyo (JP)
Filed on Mar. 5, 2018, as Appl. No. 15/911,731.
Application 15/911,731 is a continuation of application No. PCT/JP2016/075896, filed on Sep. 2, 2016.
Claims priority of application No. 2015-175112 (JP), filed on Sep. 4, 2015.
Prior Publication US 2018/0195103 A1, Jul. 12, 2018
Int. Cl. C12P 21/02 (2006.01); C12N 1/20 (2006.01); C07K 5/083 (2006.01); C07K 5/093 (2006.01); C12N 9/00 (2006.01); C12N 15/09 (2006.01); C12R 1/19 (2006.01)
CPC C12P 21/02 (2013.01) [C07K 5/0806 (2013.01); C07K 5/0819 (2013.01); C12N 1/205 (2021.05); C12N 9/93 (2013.01); C12N 15/09 (2013.01); C12R 2001/19 (2021.05); C12Y 603/02002 (2013.01); C12Y 603/02003 (2013.01)] 14 Claims
 
1. A modified, Escherichia coli bacterium,
wherein the bacterium has a deleted ybdK gene that encodes γ-glutamylglycine synthetase that is at least 90% identical to the amino acid sequence of SEQ ID NO: 16, wherein the bacterium has a reduced γ-glutamylglycine synthetase activity compared to a non-modified bacterium,
wherein the bacterium has a deleted gshA gene that encodes a γ-glutamylcysteine synthetase that is at least 90% identical to the amino acid sequence of SEQ ID NO: 24,
wherein the bacterium has a reduced γ-glutamylcysteine synthetase activity compared to a non-modified bacterium,
wherein the bacterium has a deleted gene that encodes a γ-glutamyltransferase that is at least 90% identical to the amino acid sequence of SEQ ID NO: 26, wherein the bacterium has a reduced γ-glutamyltransferase activity compared to a non-modified bacterium,
wherein the bacterium has a gene encoding γ-glutamylvaline synthetase, wherein the γ-glutamylvaline synthetase has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 24 mutated with a substitution at a position selected from L135, Q144, Y241, N243, and Y300, or wherein the γ-glutamylvaline synthetase has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 18, 20, or 22,
wherein the γ-glutamylvaline synthetase has a ratio of γ-glutamylvaline synthetase activity to γ-glutamylglycine synthetase activity of at least 3.0, and
wherein the bacterium has enhanced ability to produce γ-glutamylvaline or γ-glutamylvalylglycine compared to a non-modified bacterium.