Engineering of yeast pyruvate decarboxylase for enhanced selectivity towards carboligation

•Cys221 mutation increases the affinity of Pdc towards pyruvate.•The increased affinity for pyruvate leads to an increase carboligation capacity.•It increases the R-PAC production.•Over-expression in pdc null strain leads to maximum R-PAC production.•Mutant Pdc is able to accept exogenous acetaldehyde as substrate. The aim of the study was to increase production of (R)-PAC by altering carboligation activity of Pdc in Saccharomyces cerevisiae. Pdc1 activity was modified by over-expression as well as changing the rate of decarboxylation and carboligation by site specific mutation in Pdc1. Over-expression of mutant Pdc1 resulted in 50±2.5% increase in levels of (R)-PAC in wild-type and further 30–40% in pdc null background. The combination of mutant Pdc1 in pdc null background was successfully evaluated for production of (R)-PAC at industrial scale. This is the first report of enhancing (R)-PAC product in yeast by recombinant technology with capability of commercial production.