首页|Enhancement of C6-C10 fatty acid ethyl esters production in Saccharomyces cerevisiae CA by metabolic engineering
Enhancement of C6-C10 fatty acid ethyl esters production in Saccharomyces cerevisiae CA by metabolic engineering
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NSTL
Elsevier
C6-C10 fatty acid ethyl esters (FAEEs) are important aromatic substances in alcoholic beverages and include ethyl hexanoate (EH), ethyl octanoate (EO), and ethyl decanoate (ED). However, the biosynthesis capacity of these esters are limited because of poor precursor C6-C10 fatty acyl-CoA accumulation and low alcohol acyltransferase (AATase) activity levels in Saccharomyces cerevisiae. In this study, S. cerevisiae CA stain was selected as the host stain because of its relatively high C6-C10 FAEE productivity. Ester production was improved by metabolic engineering strategies. Acetaldehyde dehydrogenase (ALD6) and acetyl-CoA synthetase (ACS1) were overexpressed to enhance the acetyl-CoA precursor accumulation. Intracellular malonyl-CoA availability was also improved by acetyl-CoA decarboxylase (ACC1Ser659Ala, Ser1157Ala) overexpression. FAS1 and FAS2 (encoding fatty acid synthase) overexpression facilitated the condensation reactions to form C6-C10 fatty acyl-CoA from malonyl-CoA and acetyl-CoA precursors. Furthermore, the strawberry alcohol acyltransferase (SAAT) was introduced into S. cerevisiae CA to catalyze the formation of C6-C10 FAEEs from C6-C10 fatty acyl-CoA and ethanol substrates. As a consequence, the newly engineered strain achieved 25.89-, 7.27-, and 9.05-fold increases in extracellular EH (7.53 mg/L), EO (13.65 mg/L) and ED (13.87 mg/L) levels, respectively, compared to the wild-type strain CA.
NSTL
Elsevier
