Modified sterol acyltransferases
AssigneeDsm IP Assets B.V.
InventorsOtto Martin Lehmann, Harald Pichler, Holly Stolterfoht
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Abstract
The present invention is related to modified sterol acyltransferase enzymes with improved activity and/or specificity towards acylation of the vitamin D3 precursor 7-dehydrocholesterol (7-DHC) to be used in biotechnological production of vitamin D3. The invention further relates to a host strain expressing said modified enzymes and their use in a process for production of vitamin D3 or derivatives and/or metabolites thereof.
Claims19
A modified enzyme having sterol acyltransferase activity, comprising at least 99% identity to SEO ID NO:
- 1 and one or more amino acid substitution(s) at (a) position(s) corresponding to residues selected from the group consisting of 11, 281, 366, 442, 551, 554, 572, 624, 626, 627, and 636 in the polypeptide according to SEQ ID NO:
- 1, wherein the one or more amino acid substitution(s) is/are selected from the group consisting of:
- L281I,
- H554Q,
- F624L,
- G627D,
- E11G-F624L,
- E11G-G627D,
- I442V-G627D,
- I442V-L626F,
- H551Y-F572L,
- H554Q-F572L,
- E11G-D366V-C636S,
- I442V-F624L-L626F,
- I442V-L626F-G627D,
- H554Q-F572L-F624L,
- H554Q-F572L-G627D,
- E11G-D366V-F624L-C636S,
- E11G-D366V-G627D-C636S,
- E11G-D366V-I442V-F624L-C636S, and
- E11G-D366V-I442V-G627D-C636S.
A modified enzyme having sterol acyltransferase activity, comprising the amino acid sequence of SEQ ID NO:
- 1 and one or more amino acid substitution(s) at (a) position(s) corresponding to residues selected from the group consisting of 11, 281, 366, 442, 551, 554, 572, 624, 626, 627, and 636 in the polypeptide according to SEQ ID NO:
- 1, wherein the one or more amino acid substitution(s) is/are selected from the group consisting of:
- L281I,
- H554Q,
- F624L,
- G627D,
- E11G-F624L,
- E11G-G627D,
- I442V-G627D,
- I442V-L626F,
- H551Y-F572L,
- H554Q-F572L,
- E11G-D366V-C636S,
- I442V-F624L-L626F,
- I442V-L626F-G627D,
- H554Q-F572L-F624L,
- H554Q-F572L-G627D,
- E11G-D366V-F624L-C636S,
- E11G-D366V-G627D-C636S,
- E11G-D366V-I442V-F624L-C636S, and
- E11G-D366V-I442V-G627D-C636S.
Dependencies are inferred from the claim text.