Patent classifications
C12Y208/01006
BIOTIN SYNTHASES FOR EFFICIENT PRODUCTION OF BIOTIN
A recombinant microorganism includes a transgene encoding a polypeptide of a Type II biotin synthase, wherein a holo-protein of the Type II biotin synthase comprises per polypeptide chain a first [4Fe—4S] cluster (radical SAM (RS) cluster) coordinated to a CxxxCxxC motif in the polypeptide chain and a second [4Fe—4S] cluster. The Type II biotin synthase contains a serine to cysteine swap in its holo-protein amino acid sequence, that is the amino acid at the position corresponding to Ser-43 in the E. coli K12 Type I biotin synthase holo-protein is a Cysteine and the amino acid corresponding to Cys-97 in the E. coli K12 Type I biotin synthase holo-protein is a Serine. A method for producing biotin includes cultivating the recombinant microorganism in a growth medium to produce a culture; and recovering biotin from the culture.
CELL FACTORY HAVING IMPROVED IRON-SULFUR CLUSTER DELIVERY
The invention provides a genetically modified bacterial cell capable of improved iron-sulfur cluster delivery, characterized by a modified gene encoding a mutant Iron Sulfur Cluster Regulator (IscR) as well as one or more transgenes encoding polypeptides that enhance the biosynthesis of either biotin, lipoic acid or thiamine. The invention provides a method for producing either biotin, lipoic acid or thiamine using the genetically modified bacterium of the invention; as well as for the use of the genetically modified bacterial cell for either biotin, lipoic acid or thiamine production.
CELL FACTORIES FOR IMPROVED PRODUCTION OF COMPOUNDS AND PROTEINS DEPENDENT ON IRON SULFUR CLUSTERS
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.