Engineering of acetyl-CoA metabolism in yeast
AssigneeBiopetrolia AB
InventorsJens Nielsen, Verena Siewers, Anastasia Krivoruchko, Yiming Zhang, Zongjie Dai
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Abstract
The invention relates to engineering of acetyl-CoA metabolism in yeast and in particular to production of acetyl-CoA in a non-ethanol producing yeast lacking endogenous gene(s) encoding pyruvate decarboxylase and comprising a heterologous pathway for synthesis of cytosolic acetyl-CoA.
Claims9
A yeast comprising at least one heterologous pathway for synthesis of cytosolic acetyl-Coenzyme A (CoA), the at least one heterologous pathway comprising heterologous genes encoding a respective enzyme involved in synthesis of acetyl-CoA, with the proviso that the heterologous genes are not a heterologous gene encoding a pyruvate formate lyase, wherein the heterologous genes comprise:
- at least one heterologous gene encoding a cytosolic pyruvate dehydrogenase complex selected from the group consisting of genes encoding an Escherichia coli cytosolic pyruvate dehydrogenase complex, genes encoding a Saccharomyces cerevisiae pyruvate dehydrogenase complex but lacking mitochondrial target signal (MTS), genes encoding an Azotobacter vinelandii pyruvate dehydrogenase complex, and genes encoding an Enterococcus faecalis pyruvate dehydrogenase complex;
- and
- at least one heterologous gene encoding respective enzymes involved in attachment and activation of lipoyl groups to said cytosolic pyruvate dehydrogenase complex selected from the group consisting of genes encoding Escherichia coli lipoic acid synthetase and/or lipoate-protein ligase, and genes encoding Saccharomyces cerevisiae LIP2, LIP3, LIP5 and GCV3, each of which is lacking MTS.
A method of producing a yeast suitable for the production of cytosolic acetyl-Coenzyme A (CoA) comprising:
- introducing, into said yeast, at least one heterologous pathway for synthesis of cytosolic acetyl-CoA, said at least one heterologous pathway comprising heterologous genes encoding a respective enzyme involved in synthesis of acetyl-CoA, with the proviso that said heterologous genes are not a heterologous gene encoding a pyruvate formate lyase,
- wherein the heterologous genes comprise:
- at least one heterologous gene encoding a cytosolic pyruvate dehydrogenase complex selected from the group consisting of genes encoding an Escherichia coli cytosolic pyruvate dehydrogenase complex, genes encoding a Saccharomyces cerevisiae pyruvate dehydrogenase complex but lacking mitochondrial target signal (MTS), genes encoding an Azotobacter vinelandii pyruvate dehydrogenase complex, and genes encoding an Enterococcus faecalis pyruvate dehydrogenase complex;
- and
- at least one heterologous gene encoding respective enzymes involved in attachment and activation of lipoyl groups to said cytosolic pyruvate dehydrogenase complex selected from the group consisting of genes encoding Escherichia coli lipoic acid synthetase and/or lipoate-protein ligase, and genes encoding Saccharomyces cerevisiae LIP2, LIP3, LIP5 and GCV3, each of which is lacking MTS.
Dependencies are inferred from the claim text.