Genetically modified bacterial cell factory for thiamine production
AssigneeBiosynthia Aps
InventorsLuisa Gronenberg, Bo Salomonsen, Matteo Ferla, Hans Jasper Genee
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Abstract
The invention provides a genetically modified bacterium for production of thiamine; where the bacterium is characterized by a transgene encoding a thiamine monophosphate phosphatase (TMP phosphatase having EC 3.1.3.-) as well as transgenes encoding polypeptides that catalyze steps in the thiamine pathway. The genetically modified bacterium is characterized by enhanced synthesis and release of thiamine into the extracellular environment. The invention further provides a method for producing thiamine using the genetically modified bacterium of the invention; as well as the use of the genetically modified bacterium for extracellular thiamine production.
Claims16
A genetically modified bacterium for production of un-phosphorylated thiamine;
- wherein said bacterium is characterized by having transgenes encoding:
- a) a polypeptide having thiamine mono-phosphate phosphatase activity (E.C. 3.1.3), wherein the amino acid sequence of said polypeptide has at least 80% sequence identity to a sequence selected from the group consisting of SEQ ID NO:
- 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72 and 74;
- b) a polypeptide having 4-amino-5-hydroxymethyl-2-methylpyrimidine phosphate (HMP-P) synthase activity (E.C. 4.1.99.17);
- c) a polypeptide having thiamine phosphate synthase activity (E.C.2.5.1.3);
- d) a polypeptide having ThiS adenylyltransferase activity (E.C.2.7.7.73);
- e) a polypeptide having ThiS sulfur-carrier activity;
- f) a polypeptide having thiazole synthase activity (E.C.2.8.1.10);
- g) a polypeptide having 2-iminoacetate synthase activity (E.C.4.1.99.19) or having glycine oxidase activity (E.C.1.4.3.19);
- and
- h) a polypeptide having phosphohydroxymethylpyrimidine kinase activity (E.C.2.7.4.7), wherein the genus of the bacterium is Escherichia.
A method for producing un-phosphorylated thiamine comprising the steps of:
- a) introducing a genetically modified Escherichia bacterium comprising a transgene encoding a polypeptide having thiamine mono-phosphate phosphatase activity (E.C3.1.3) into a growth medium to produce a culture;
- b) cultivating the culture;
- and
- c) recovering thiamine produced by said culture, and optionally purifying the recovered thiamine.
Dependencies are inferred from the claim text.