Application of Acetyl-CoA Synthetase from Methanothermobacter thermautotrophicus to Non-Native Substrates

Publication date: Available online 14 May 2019Source: Enzyme and Microbial TechnologyAuthor(s): Louis M.M. Mouterde, Jon D. StewartAbstractThe substrate selectivity of the Trp416Gly mutant of Methanothermobacter thermautotrophicus acetyl-CoA synthetase (Trp416Gly MT-ACS1) was explored. The goal was to identify its substrate range, particularly for functionalized carboxylic acid substrates that would allow post-synthesis functionalization of CoA thioesters or downstream products using metathesis or Click chemistry. Relative activities were determined by in situ formation of acyl-hydroxamate iron (III) complexes. Trp416Gly MT-ACS1 showed good activities for saturated straight chain carboxylic acids from C2 to C8, for ω-alkenyl straight chain carboxylic acids from C4 to C7 and for ω-alkynyl straight chain carboxylic acids from C5 to C7. Carboxylic acids showing ≥20% conversion in screening reactions were used in preparative conversions that completely consumed the added CoASH.
Source: Enzyme and Microbial Technology - Category: Biotechnology Source Type: research