US 11,851,461 B2
Cell factory having improved iron-sulfur cluster delivery
Hans Jasper Genee, Copenhagen N (DK); Anne Pihl Bali, Copenhagen NV (DK); and Nils Myling-Petersen, Copenhagen Ø (DK)
Assigned to BIOSYNTIA APS, Copenhagen Ø (DK)
Appl. No. 16/630,203
Filed by BIOSYNTIA APS, Copenhagen Ø (DK)
PCT Filed Jul. 12, 2018, PCT No. PCT/EP2018/068989
§ 371(c)(1), (2) Date Jan. 10, 2020,
PCT Pub. No. WO2019/012058, PCT Pub. Date Jan. 17, 2019.
Claims priority of application No. 17181503 (EP), filed on Jul. 14, 2017.
Prior Publication US 2023/0192778 A1, Jun. 22, 2023
Int. Cl. C12N 9/00 (2006.01); C07K 14/245 (2006.01); C12N 9/10 (2006.01); C12N 9/88 (2006.01); C12N 15/70 (2006.01); C12N 15/77 (2006.01)
CPC C07K 14/245 (2013.01) [C12N 9/13 (2013.01); C12N 9/88 (2013.01); C12N 9/93 (2013.01); C12N 15/70 (2013.01); C12N 15/77 (2013.01)] 18 Claims
 
1. A genetically modified bacterium wherein the bacterium has an enhanced production of biotin or lipoic acid or thiamine; wherein said bacterium comprises:
a. a genetically modified endogenous iscR gene encoding a mutant
IscR polypeptide, wherein the amino acid sequence of said mutant IscR polypeptide has at least 80% amino acid sequence identity to a sequence selected from the group consisting of SEQ ID No: 2, 4, 6, 8, 10, 12 and 14, and wherein said amino acid sequence has at least one amino acid substitution selected from the group consisting of:
i) L15X, C92X, C98X, C104X, and H 107X; wherein X is any amino acid other than the corresponding amino acid residue in SEQ ID No.: 2, 4, 6, 8, 10, 12 and 14; and
b. at least one transgene encoding a polypeptide selected from the group consisting of:
ii) a polypeptide having biotin synthase activity (EC 2.8.1.6),
iii) a polypeptide having lipoic acid synthase activity (EC 2.8.1.8),
iv). a polypeptide having HMP-P synthase activity (EC 4.1.99.17), and
v). a polypeptide having tyrosine lyase activity (EC 4.1.99.19).