Investigations of substrate specificity of lipase and esterase from Triticum aestivum and their Michaelis-Menten kinetics

Abstract

Enzymatic screening is screening an enzyme library to identify those that possess enzymatic activity towards defined substrates. In this work we screened a lipase and an esterase from Triticum aestivum – most significant cereal crop. The study aimed to investigate whether long-chain or short-chain fatty acids have higher processing rate by esterase and lipase from Triticum aestivum. In the screening, pH values were varied. Moreover, as substrates, several esters with varying length of fatty acids were used. The lipase and esterase showed higher activity towards substrates with a longer carbon chain. Enzyme activity was measured photometrically by using para-nitrophenol. Inhibitory properties of diisopropylfluorophosphate in relation to the esterase were tested. It was found that diisopropylfluorophosphate acts as a non-competitive inhibitor. pH optimum of pancreatin was determined, the value of which is 7.