Linalool synthase variants are disclosed. The variants preferably comprise an amino acid sequence having one or more amino acid substitutions. Variants producing a greater linalool yield and/or higher linalool:nerolidol ratio compared with the corresponding wild type linalool synthase are also disclosed. A method comprising the conversion of geranyl pyrophosphate to linalool using a linalool synthase variant is also disclosed.

The present invention relates to a process of producing a monoterpene and/or derivatives thereof. The process comprises the steps of: a) providing a host microorganism genetically engineered to express a bacterial monoterpene synthase (mTS); and b) contacting geranyl pyrophosphate (GPP) with said bacterial mTS to produce said monoterpene and/or derivatives thereof. The present invention also relates to a microorganism for use in producing a monoterpene and/or derivatives thereof and a recombinant microor-ganism adapted to conduct the step of converting geranyl pyrophosphate (GPP) into a monoterpene and/or derivatives thereof by ex-pression of a bacterial mTS. It was shown to produce 1,8 cineole using 1,8 cineole synthase and to produce linalool using linalool synthase, both from Streptomyces clavuligerus.

The present invention provides a composition containing linalool, wherein the composition contains a high amount of either enantiomer R-linalool or S-linalool, and has a high content rate of linalool. The present invention also provides a production method for producing the composition. The present invention further provides a composition containing volatile components including linalool, in which a content of linalool in a total content of the volatile components in the composition is 60% or more, and the linalool is present as R-linalool or S-linalool in an amount of 50% or more of the enantiomer, and a production method therefor.

The present invention describes an efficient method for producing linalool. The present invention provides a method for producing linalool, the method including culturing a microorganism expressing linalool synthase in a culture medium to produce linalool. The present invention also describes a microorganism able to express linalool synthase and efficiently produce linalool.

The present invention relates to a process of producing a monoterpene and/or derivatives thereof. The process comprises the steps of: a) providing a host microorganism genetically engineered to express a bacterial monoterpene synthase (mTS); and b) contacting geranyl pyrophosphate (GPP) with said bacterial mTS to produce said monoterpene and/or derivatives thereof. The present invention also relates to a microorganism for use in producing a monoterpene and/or derivatives thereof and a recombinant microorganism adapted to conduct the step of converting geranyl pyrophosphate (GPP) into a monoterpene and/or derivatives thereof by expression of a bacterial mTS. It was shown to produce 1,8 cineole using 1,8 cineole synthase and to produce linalool using linalool synthase, both from Streptomyces clavuligerus.

The present invention relates to an alcohol acyl transferase which is capable of esterifying a tertiary monoterpene alcohol such that at least 30% by mass of said tertiary monoterpene alcohol is esterified, preferably within 36 h, 24 h, 18 h, 12 h, 6 h, 3 h, 2 h, 1 h, 45 min or 30 min, more preferably in a microbial cell. The invention further relates to a nucleic acid comprising a nucleic acid sequence encoding the alcohol acyl transferase of the invention, or a complementary sequence thereof, and a vector or gene construct comprising the nucleic acid of the invention. Further provided by the present invention is a host cell comprising the vector or gene construct of the invention, and a transgenic non-human organism comprising the nucleic acid of the invention, the vector or gene construct of the invention, or the host cell of the invention. The invention also concerns a method for preparing a monoterpene ester, comprising esterifying a monoterpene alcohol to a monoterpene ester, in the presence of an alcohol acyl transferase of the invention. Specifically, it provides a method for preparing linalyl acetate, comprising esterifying linalool to linalyl acetate, in the presence of an alcohol acyl transferase of the invention. The invention further pertains to the use of the alcohol acyl transferase of the invention, the nucleic acid of the invention, the vector or gene construct of the invention, the host cell of the invention, or the transgenic non-human organism of the invention (i) for heterologous reconstitution of a terpene biosynthetic pathway; (ii) for producing an industrial product, preferably a flavour or fragrance, a biofuel, a fuel composition, a fuel compound, e.g., a blowing agent for diesel fuel compositions, a pesticide, an insect repellent or an antimicrobial; (ill) for producing an aliphatic and/or aromatic monoterpene ester from a monoterpene alcohol, preferably from a tertiary monoterpene alcohol; (iv) for detoxifying a monoterpene alcohol in a microorganism, thereby increasing monoterpene production in said microorganism; (v) in combination with a GPP synthase and/or S- or R-linalool synthase; (vi) for increasing the beneficial effects of acetylation in that the hydrophobic acetate partitions more readily go into an organic phase, as compared to the monoterpene alcohol; (vii) for expressing the alcohol acyl transferase of the invention such that the ratio of monoterpene acetate to monoterpene alcohol is greater than 5:1 or 10:1 or (viii) in a microbial production system for monoterpene esters. The invention also provides a kit comprising the alcohol acyl transferase of the invention, the nucleic acid of the invention, the vector or gene

The present invention relates to a process of producing a monoterpene and/or derivatives thereof. The process comprises the steps of: a) providing a host microorganism genetically engineered to express a bacterial monoterpene synthase (mTS); and b) contacting geranyl pyrophosphate (GPP) with said bacterial mTS to produce said monoterpene and/or derivatives thereof. The present invention also relates to a microorganism for use in producing a monoterpene and/or derivatives thereof and a recombinant microor-ganism adapted to conduct the step of converting geranyl pyrophosphate (GPP) into a monoterpene and/or derivatives thereof by ex-pression of a bacterial mTS. It was shown to produce 1,8 cineole using 1,8 cineole synthase and to produce linalool using linalool synthase, both from Streptomyces clavuligerus.

The present invention provides a composition containing linalool, wherein the composition contains a high amount of either enantiomer R-linalool or S-linalool, and has a high content rate of linalool. The present invention also provides a production method for producing the composition. The present invention further provides a composition containing volatile components including linalool, in which a content of linalool in a total content of the volatile components in the composition is 60% or more, and the linalool is present as R-linalool or S-linalool in an amount of 50% or more of the enantiomer, and a production method therefor.

The present invention describes an efficient method for producing linalool. The present invention provides a method for producing linalool, the method including culturing a microorganism expressing linalool synthase in a culture medium to produce linalool. The present invention also describes a microorganism able to express linalool synthase and efficiently produce linalool.

Linalool synthase variants are disclosed. The variants preferably comprise an amino acid sequence having one or more amino acid substitutions. Variants producing a greater linalool yield and/or higher linalool:nerolidol ratio compared with the corresponding wild type linalool synthase are also disclosed. A method comprising the conversion of geranyl pyrophosphate to linalool using a linalool synthase variant is also disclosed.