Methods for production of oxygenated terpenes
The present disclosure relates to methods for producing oxygenated terpenoids, and preparation of compositions and formulations thereof. Polynucleotides, derivative enzymes, and host cells for use in such methods are also provided.
1 . A method of making a product for providing oxygenated terpenes, comprising: obtaining a formulation by contacting valencene with Stevia rebaudiana Kaurene Oxidase (SrKO) or an SrKO derivative comprising a sequence having at least 70% sequence identity to SEQ ID NO: 37, SEQ ID NO: 38, or SEQ ID NO: 55, and having valencene oxidizing activity in an in vitro or in vivo system to produce an oxygenated product comprising nootkatone, α-nootkatol, and β-nootkatol, and incorporating the formulation into a consumer product, an industrial product, a flavor product, a fragrance product, a cosmetic product, a cleaning product, a detergent product, a soap product, or a pest control product.
2 . The method of claim 1 , wherein the flavor product is a beverage, a chewing gum, a candy, a natural flavor, an artificial flavor, or a flavor additive.
3 . The method of claim 1 , wherein obtaining the formulation comprises contacting valencene with Stevia rebaudiana Kaurene Oxidase (SrKO) or an SrKO derivative comprising a sequence having at least 70% sequence identity to SEQ ID NO: 37, SEQ ID NO: 38, or SEQ ID NO: 55, and having valencene oxidizing activity in an in vivo or in vitro system to yield an oxygenated product.
4 . The method of claim 1 , further comprising processing the oxygenated product by fractional distillation to yield two or more fractions of the oxygenated product.
5 . The method of claim 4 , further comprising blending the two or more fractions of the oxygenated product.
6 . The method of claim 1 , further comprising adding an unoxygenated product to the oxygenated product, one or more fractions of the oxygenated product, or the formulation.
7 . The method of claim 6 , wherein the unoxygenated product comprises valencene.
8 . The method of claim 7 , wherein the valencene is present in the formulation in an amount ranging from about 5% to about 10% (w/w).
9 . The method of claim 1 , wherein the formulation further comprises one or more non-sesquiterpene components or sesquiterpene components.
10 . The method of claim 9 , wherein the one or more non-sesquiterpene components or sesquiterpene components is selected from Table 8A, Table 8B, and Table 9.