Methods for production of novel diterpene scaffolds

A method for synthesizing a terpene comprising incubating a terpene precursor of a terpene of formula I, II, or III, with an enzyme with at least 95% sequence identity to the amino acid sequence of SEQ ID NO:9, wherein the terpene of formula I, II, or III is:

• wherein
• each R1 can separately be hydrogen or lower alkyl;
• R2 can be hydrogen, lower alkyl, hydroxy, a bond to an adjacent ring carbon, or form a C4-C6 cycloheteroalkyl with R3;
• R3 can be a branched C5-C6 alkyl with 0-2 double bonds, can form a C4-C6 cycloheteroalkyl with R2;
• can form a cycloalkyl with R4, or can form a cycloheteroalkyl ring with R4, wherein the C5-C6 alkyl can optionally have one hydroxy, phosphate or diphosphate substituent, and wherein each cycloalkyl or cycloheteroalkyl ring can have 0-2 double bonds, and each cycloalkyl or cycloheteroalkyl ring can have 0-2 alkyl or 0-2 alkene substituents;
• R4 can be hydrogen, lower alkyl, lower alkene, hydroxy, a carbon bonded to R9, an oxygen bonded to R9, form a cycloalkyl ring with R3, or form a cycloheteroalkyl ring with R3, wherein each cycloalkyl ring or cycloheteroalkyl ring can have 0-2 double bonds, and each cycloalkyl ring or cycloheteroalkyl ring can have 0-2 alkyl or 0-2 alkene substituents;
• R5 can be hydrogen, hydroxy, lower alkyl, a lower alkene, a bond with an adjacent carbon, form a cycloalkyl ring with a ring atom of a ring formed by R3 and R4, wherein the cycloalkyl ring can have 0-2 double bonds, and the cycloalkyl ring can have 0-2 alkyl or 0-2 alkene substituents;
• each R6 can separately be hydrogen, lower alkyl, lower alkene, or form a bond with an adjacent carbon;
• R7 can be lower alkyl, lower alkene, or form a cycloalkyl ring with a R5,
• R8 can be lower alkyl, hydroxy, phosphate, diphosphate, or form a bond with an adjacent carbon;
• and
• R9 can be hydrogen, lower alkyl, lower alkene, ═CH2, hydroxy, phosphate, diphosphate, form a bond with an adjacent carbon, form a cycloalkyl ring with R4, or form a cycloheteroalkyl ring with R4, wherein each cycloalkyl ring or cycloheteroalkyl ring can have 0-2 double bonds, and each cycloalkyl ring or cycloheteroalkyl ring can have 0-2 alkyl or 0-2 alkene substituents.