US 2022/0127311 A1 · CELL FACTORIES FOR IMPROVED PRODUCTION OF COMPOUNDS AND PROTEINS DEPENDENT ON IRON SULFUR CLUSTERS

Abstract

The invention relates to a genetically modified prokaryotic cell capable of improved iron-sulfur cluster delivery, characterized by a modified gene encoding a mutant Iron Sulfur Cluster Regulator (IscR) and one or more transgenes or upregulated endogenous genes encoding iron-sulfur (Fe—S) cluster polypeptides or proteins that catalyze complex radical-mediated molecular rearrangements, electron transfer, radical or non-redox reactions, sulfur donation or perform regulatory functions. The prokaryotic cells are characterized by enhanced activity of these iron-sulfur (Fe—S) cluster polypeptides, enhancing their respective functional capacity, and facilitating enhanced yields of compounds in free and protein-bound forms, including heme, hemoproteins, tetrapyrroles, B vitamins, amino acids, δ-aminolevulinic acid, biofuels, isoprenoids, pyrroloquinoline quinone, ammonia, indigo, or their precursors, whose biosynthesis depends on their activity. The invention further relates to a method for producing said compounds or their precursors using the genetically modified prokaryotic cell of the invention, and the use of the genetically modified prokaryotic cell.

Claims28

Dependent claims

A genetically modified prokaryotic cell comprising:

a) a transgene or a genetically modified iscR gene encoding a mutant IscR polypeptide;
and
b) transgenes or endogenous genes encoding at least one Fe—S cluster polypeptide, wherein the at least one FE—S cluster polypeptide is not any one of i. biotin synthase (EC:
2.8.1.6), ii. lipoic acid synthase (EC:
2.8.1.8), iii. HMP-P synthase (EC:
4.1.99.17), and vi. tyrosine lyase (EC:
4.1.99.19);
and
wherein the endogenous genes are operably linked to genetically modified regulatory sequences capable of enhancing expression of said endogenous genes, and wherein the mutant IscR polypeptide as compared to a corresponding non-mutant IscR polypeptide has an increased apoprotein:holoprotein ratio in the cell;
and wherein the production of at least one compound resulting from the catalytic activity of the at least one Fe—S cluster polypeptide is enhanced when compared to a prokaryotic cell comprising the genetically unmodified iscR gene and the transgenes or the endogenous genes encoding at least one Fe—S polypeptide.

7 more elements

Use of a genetically modified gene encoding a mutant iscR polypeptide to increase production of at least one compound resulting from the catalytic activity of at least one Fe—S cluster polypeptide in a genetically modified prokaryotic cell as compared to a prokaryotic cell comprising the genetically unmodified iscR gene, wherein said Fe—S cluster polypeptide is not any one of biotin synthase (EC:

2.8.1.6), lipoic acid synthase (EC:
2.8.1.8), HMP-P synthase (EC:
4.1.99.17), and tyrosine lyase (EC:
4.1.99.19), wherein the mutant IscR polypeptide as compared to a non-mutant IscR polypeptide has an increased apoprotein:holoprotein ratio in the cell.

1 more element

Dependencies are inferred from the claim text.

Cell factories for improved production of compounds and proteins dependent on iron sulfur clusters

Life of this patent

Cell factories for improved production of compounds and proteins dependent on iron sulfur clusters

Life of this patent

Abstract

Claims28