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US 11,505,811 B2Granted
Method for the preparation of 2,4-dihydroxybutyrate
AssigneeAdisseo France S.A.S.
InventorsThomas Walther, Hélène Cordier, Clémentine Dressaire, Jean Marie Francois, Robert Huet
Life of this patent
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Abstract
A method for the preparation of 2,4-dihydroxybutyric acid from homoserine includes a first step of conversion of the primary amino group of homoserine to a carbonyl group to obtain 2-oxo-4-hydroxybutyrate, and a second step of reduction of the obtained 2-oxo-4-hydroxybutyrate (OHB) to 2,4-dihydroxybutyrate.
Claims13
A method for the preparation of 2,4-dihydroxybutyrate (2,4-DHB) from homoserine comprising:
- deaminating homoserine to form 2-oxo-4-hydroxybutyrate (OHB), where the deamination of homoserine is catalyzed by an enzyme having homoserine transaminase activity, wherein
- the enzyme having homoserine transaminase activity is produced via a transformed host microorganism that comprises a first chimeric gene including a first nucleic acid sequence encoding the enzyme having homoserine transaminase activity for converting the primary amino group of homoserine to a carbonyl group to obtain OHB, and
- the enzyme having homoserine transaminase activity is selected from the group consisting of aspC from Escherichia coli (SEQ ID NO:
- 64), ilvE from Escherichia coli (SEQ ID NO:
- 60), bcaT from Lactococcus lactis (SEQ ID NO:
- 68), tyrB from Escherichia coli (SEQ ID NO:
- 62), araT from Lactococcus lactis (SEQ ID NO:
- 66), and ARO8 from Saccharomyces cerevisiae (SEQ ID NO:
- 70), or is selected from any sequence sharing a sequence identity of at least 90% with at least one of the sequences of said enzymes having homoserine transaminase activity;
- and
- reducing the OHB to form 2,4-DHB, where the reduction of OHB is catalyzed by an enzyme having OHB reductase activity, wherein
- the enzyme having OHB reductase activity is produced via the transformed host microorganism, which further comprises a second chimeric gene including a second nucleic acid sequence encoding the enzyme having OHB reductase activity for reducing OHB to 2,4-DHB, and
- the enzyme having OHB reductase activity is selected from the group consisting of (L)-malate dehydrogenase from Escherichia coli (SEQ ID NO:
- 2), (D)-lactate dehydrogenase from Escherichia coli (SEQ ID NO:
- 4), (L)-lactate dehydrogenase from Lactococcus lactis (SEQ ID NO:
- 6), (L)-lactate dehydrogenase from Bacillus subtilis (SEQ ID NO:
- 8), (L)-lactate dehydrogenase from Geobacillus stearothermophilus (SEQ ID NO:
- 10), the two isoforms of (L)-lactate dehydrogenase from Oryctalagus cuniculus (SEQ ID NO:
- 12 and SEQ ID NO:
- 14), or is selected from any sequence sharing a sequence identity of at least 90% with at least one of said enzymes having OHB reductase activity.
17 more elements
A modified microorganism for the preparation of 2,4-dihydroxybutyrate (2,4-DHB) from homoserine via a two-step pathway comprising:
- deaminating homoserine to form 2-oxo-4-hydroxybutyrate (OHB), where the deamination of homoserine is catalyzed by an enzyme having homoserine transaminase activity selected from the group consisting of aspC from Escherichia coli (SEQ ID NO:
- 64), ilvE from Escherichia coli (SEQ ID NO:
- 60), bcaT from Lactococcus lactis (SEQ ID NO:
- 68), tyrB from Escherichia coli (SEQ ID NO:
- 62), araT from Lactococcus lactis (SEQ ID NO:
- 66), and ARO8 from Saccharomyces cerevisiae (SEQ ID NO:
- 70), or is selected from any sequence sharing a sequence identity of at least 90% with at least one of the sequences of said enzymes having homoserine transaminase activity, and
- reducing the OHB to form 2,4-DHB, where the reduction of OHB is catalyzed by an enzyme having OHB reductase activity, wherein the enzyme having OHB reductase activity is selected from the group consisting of (L)-malate dehydrogenase from Escherichia coli (SEQ ID NO:
- 2), (D)-lactate dehydrogenase from Escherichia coli (SEQ ID NO:
- 4), (L)-lactate dehydrogenase from Lactococcus lactis (SEQ ID NO:
- 6), (L)-lactate dehydrogenase from Bacillus subtilis (SEQ ID NO:
- 8), (L)-lactate dehydrogenase from Geobacillus stearothermophilus (SEQ ID NO:
- 10), and the two isoforms of (L)-lactate dehydrogenase from Oryctalagus cuniculus (SEQ ID NO:
- 12 and SEQ ID NO:
- 14), or is selected from any sequence sharing a sequence identity of at least 90% with at least one of said enzymes having homoserine transaminase activity;
- wherein
- the modified microorganism is a host microorganism that has been transformed to enhance production of 2,4-DHB compared to a non-transformed host microorganism, the transformed host microorganism comprising:
- a first chimeric gene including a first nucleic acid sequence encoding the enzyme having homoserine transaminase activity for converting the primary amino group of homoserine to a carbonyl group to obtain OHB, and
- a second chimeric gene including a second nucleic acid sequence encoding the enzyme having OHB reductase activity for reducing OHB in 2,4-DHB.
16 more elements
Dependencies are inferred from the claim text.