The invention relates to microbial terpene production. Known methods for microbial production of terpenes are mostly based on the direct conversion of sugars. Therefore alternative substrates, in particular alternative carbon sources, for use in microbial terpene production were desirable. The invention relates to a methylotrophic bacterium containing recombinant DNA coding for at least one polypeptide with enzymatic activity for heterologous expression in said bacterium, wherein said at least one polypeptide with enzymatic activity is selected from the group consisting an enzyme of a heterologous mevalonate pathway, a heterologous terpene synthase and optionally a heterologous synthase of a prenyl diphosphate precursor. The invention further relates in particular to a method for de novo microbial synthesis of sesquiterpenes or diterpenes from methanol and/or ethanol.

The present invention provides for a genetically modified host cell capable of producing isopentenol and/or 3-methyl-3-butenol, comprising (a) an increased expression of phosphomevalonate decarboxylase (PMD) (b) an increased expression of a phosphatase capable of converting isopentenol into 3-methyl-3-butenol, (c) optionally the genetically modified host cell does not express, or has a decreased expression of one or more of NudB, phosphomevalonate kinase (PMK), and/or PMD, and (d) optionally one or more further enzymes capable of converting isopentenol and/or 3-methyl-3-butenol into a third compound, such as isoprene.

The invention provides a method for producing an isoprenoid or a precursor thereof by microbial fermentation. Typically, the method involves culturing a recombinant bacterium in the presence of a gaseous substrate whereby the bacterium produces an isoprenoid or a precursor thereof, such as mevalonic acid, isopentenyl pyrophosphate, dimethylallyl pyrophosphate, isoprene, geranyl pyrophosphate, farnesyl pyrophosphate, and/or pinene. The bacterium may comprise one or more exogenous enzymes, such as enzymes in mevalonate, DXS, isoprenoid alcohol, or terpene biosynthesis pathways.

The present disclosure provides genetically modified host cells that produce a cannabinoid, a cannabinoid derivative, a cannabinoid precursor, or a cannabinoid precursor derivative. The present disclosure provides methods of synthesizing a cannabinoid, a cannabinoid derivative, a cannabinoid precursor, or a cannabinoid precursor derivative.

The invention features compositions and methods for the increased production of mevalonate, isoprene, isoprenoid precursor molecules, and/or isoprenoids in microorganisms via the heterologous expression of the mvaE and mvaS genes from the organisms Listeria grayi DSM 20601, Enterococcus faecium, Enterococcus gallinarum EG2, and Enterococcus casseliflavus.

This application describes methods, including non-naturally occurring methods, for biosynthesizing unsaturated pentahydrocarbons, such as isoprene and intermediates thereof, via the mevalonate pathway, as well as non-naturally occurring hosts for producing isoprene.

Described is a method for the production of 3-hydroxy-3-methylbutyric acid by enzyme-catalyzed covalent bond formation between the carbon atom of the oxo group of acetone and the methyl group of a compound which provides an activated acetyl group. Also described are recombinant organisms which produce 3-hydroxy-3-methylbutyric acid, and related compositions and methods.

The present disclosure belongs to the field of synthetic biology and relates to a valencene synthase mutant and a valencene high-yield strain. An enzyme for synthesizing valencene is derived from Eryngium glaciale, and upon enzyme directed evolution of the enzyme, a valencene synthase mutant with improved enzyme performance is obtained, and the yield of a strain containing the mutant is 3.15 times the yield of a strain containing a wild-type synthase. The valencene synthase mutant of the present disclosure enhances the capability of synthesizing valencene by a strain, and a powerful foundation is laid for the industrial production thereof. A high-yield strain for synthesizing valencene is constructed by using the valencene synthetase mutant, and the yield of a fermentation tank reaches 12.4 g/L, which is the highest level reported to date.

The invention provides a method for producing a terpene or a precursor thereof by microbial fermentation. Typically, the method involves culturing a recombinant bacterium in the presence of a gaseous substrate whereby the bacterium produces a terpene or a precursor thereof, such as mevalonic acid, isopentenyl pyrophosphate, dimethylallyl pyrophosphate, isoprene, geranyl pyrophosphate, farnesyl pyrophosphate, and/or farnesene. The bacterium may comprise one or more exogenous enzymes, such as enzymes in mevalonate, DXS, or terpene biosynthesis pathways.

The present invention provides for a method to increase production of isoprenol by a genetically modified Pseudomonas cell, the method comprising: (a) providing a genetically modified Pseudomonas cell comprising one or more of heterologous genes encoding: MvaE, AtoB, MvaS, MK, PMD.sub.HKQ, AphA, and PhoA; and (b) culturing or growing the genetically modified Pseudomonas cell in a medium to produce isoprenol; wherein (i) the genetically modified Pseudomonas cell is deleted, knocked out, or reduced in expression of one or more of the following endogenous genes: a gene at PP_2675 locus (or a deletion of the PP_2675 locus), phaABC, mvaB, hbdH, ldhA, gntZ, ppsA, pycAB, gltA, and aceA, and/or (ii) the medium comprises one or more amino acids that reduce the catabolism of isoprenol.