Abstract
BackgroundGeranyl acetate is widely used in the fragrance and cosmetic industries, and thus has great economic value. However, plants naturally produce a mixture of hundreds of esters, and geranyl acetate is usually only present in trace amounts, which makes its economical extraction from plant sources practically impossible. As an ideal host for heterologous production of fragrance compound, the Saccharomyces cerevisiae has never been engineered to produce the esters, such as geranyl acetate.ResultsIn this study, a heterologous geranyl acetate synthesis pathway was constructed in S. cerevisiae for the first time, and a titer of 0.63 mg/L geranyl acetate was achieved. By expressing an Erg20 mutant to divert carbon flux from FPP to GPP, the geranyl acetate production increased to 2.64 mg/L. However, the expression of heterologous GPP had limited effect. The highest production of 13.27 mg/L geranyl acetate was achieved by additional integration and expression of tHMG1, IDI1 and MAF1. Furthermore, through optimizing fermentation conditions, the geranyl acetate titer increased to 22.49 mg/L.ConclusionsWe constructed a monoterpene ester producing cell factory in S. cerevisiae for the first time, and demonstrated the great potential of this system for the heterologous production of a large group of economically important fragrance compounds.
Highlights
Geranyl acetate is widely used in the fragrance and cosmetic industries, and has great economic value
Construction of a microbial cell factory by integrating the geranyl acetate biosynthetic pathway into the chromosome of S. cerevisiae As volatile esters, geranyl acetate is the essential components of fruit characteristic aroma and presents in the essential oils of various plant species
It has been reported that truncated O. basilicum geraniol synthase is very efficient in geraniol synthesis with geranyl diphosphate (GPP) as the substrate [3, 16]
Summary
Geranyl acetate is widely used in the fragrance and cosmetic industries, and has great economic value. Plants naturally produce a mixture of hundreds of esters, and geranyl acetate is usually only present in trace amounts, which makes its economical extraction from plant sources practically impossible. As an ideal host for heterologous production of fragrance compound, the Saccharomyces cerevisiae has never been engineered to produce the esters, such as geranyl acetate. Monoterpenes constitute a subclass of terpenoids [1] that are widely used as additives in the food, pharmaceutical, agrichemical and cosmetic industries, due to their strong flavor, fragrance and physiological activity [2, 3]. A number of research groups have been able to produce natural products by metabolic engineering of microbial cell factories [10, 11], most often derived from the model organisms Escherichia coli and Saccharomyces cerevisiae. Previous work has shown that engineered E. coli could produce 400 mg/L of limonene and approximately 100 mg/L of perillyl alcohol [13], and engineered yeasts were able to produce 95 μg/L of linalool [15], as well as 36.04 mg/L–2.0 g/L of geraniol [16, 17]
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