Production of manool
AssigneeFirmenich SA
InventorsMichel Schalk, Laurent Daviet, Letizia Rocci, Daniel Solis Escalante
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Abstract
Described herein are methods of producing (+)-manool, the methods including: contacting geranylgeranyl diphosphate with a copalyl diphosphate (CPP) synthase to form a (9S, 10S)-copalyl diphosphate and contacting the CPP with a sclareol synthase enzyme to form (+)-manool and derivatives thereof. Also described herein are nucleic acids encoding CPP synthases and sclareol synthases for use in the methods. Further described herein are expression vectors and non-human host organisms and cells including nucleic acids encoding a CPP synthase and a sclareol synthase as described herein.
Claims16
A method of producing (+)-manool, the method comprising:
- a) contacting geranylgeranyl diphosphate (GGPP) with a copalyl diphosphate (CPP) synthase to form a copalyl diphosphate, wherein the CPP synthase comprises
- a) an amino acid sequence having at least 90% sequence identity to SEQ ID NO:
- 14 or SEQ ID NO:
- 15;
- or
- b) an amino acid sequence having at least 90% sequence identity to SEQ ID NO:
- 17 or SEQ ID NO:
- 18;
- or
- c) an amino acid sequence having at least 90% sequence identity to SEQ ID NO:
- 20 or SEQ ID NO:
- 21;
- and
- b) contacting the CPP with a sclareol synthase to form the (+)-manool;
- and
- c) optionally isolating the (+)-manool.
A method for transforming a host cell or non-human organism, the method comprising transforming a host cell or non-human organism with a nucleic acid encoding a polypeptide having a (9S, 10S)-copalyl diphosphate synthase activity and a nucleic acid encoding a polypeptide having a sclareol synthase activity,
- wherein the polypeptide having (9S, 10S)-copalyl diphosphate activity comprises
- a) an amino acid sequence having at least 90% sequence identity to SEQ ID NO:
- 14 or SEQ ID NO:
- 15;
- or
- b) an amino acid sequence having at least 90% sequence identity to SEQ ID NO:
- 20 and SEQ ID NO:
- 21;
- and
- wherein the polypeptide having sclareol synthase activity comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:
- 4, SEQ ID NO:
- 5, SEQ ID NO:
- 23, or SEQ ID NO:
- 25.
An expression vector comprising
- a) a nucleic acid encoding a polypeptide having a (9S, 10S)-copalyl diphosphate synthase activity comprising
- a) an amino acid sequence having at least 90% sequence identity to SEQ ID NO:
- 14 or SEQ ID NO:
- 15;
- or
- b) an amino acid sequence having at least 98% sequence identity to SEQ ID NO:
- 20 or SEQ ID NO:
- 21;
- or
- b) a nucleic acid encoding a polypeptide having a (9S, 105)-copalyl diphosphate synthase activity comprising a nucleotide sequence having at least 90% sequence identity to a nucleic acid sequence as set forth in SEQ ID NO:
- 13, SEQ ID NO:
- 16, SEQ ID NO:
- 19, SEQ ID NO:
- 22, SEQ ID NO:
- 26, SEQ ID NO:
- 29, SEQ ID NO:
- 30, SEQ ID NO:
- 31, or SEQ ID NO:
- 32;
- and
- c) a nucleic acid encoding a polypeptide having a sclareol synthase activity, wherein the polypeptide having sclareol synthase activity comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:
- 4, SEQ ID NO:
- 5, SEQ ID NO:
- 23, or SEQ ID NO:
- 25;
- or
- d) a nucleic acid encoding a polypeptide having a sclareol synthase activity comprising a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:
- 6, SEQ ID NO:
- 24, SEQ ID NO:
- 27, SEQ ID NO:
- 33, or SEQ ID NO:
- 34.
Dependencies are inferred from the claim text.