Ultrasound enhanced biosynthesis of L-theanine from L-glutamine and ethylamine by recombinant γ-glutamyltranspeptidase.

A mutant library of the key amino acid residue site E387 in γ-glutamyltranspeptidase was constructed to screen the mutant enzymes with significantly improved thermal stability (E387Q). The reaction temperature of the mutant enzyme (E387Q) was 10℃ higher than that of the parent enzyme. Ultrasound-assisted synthesis of L-theanine by γ-glutamyltranspeptidase was investigated. The effects of ultrasonic power, reaction pH and substrate concentration on the enzymatic synthesis of L-theanine were studied by the response surface method. The results showed that the optimal process conditions are ultrasonic power of 100 W, reaction pH of 9, substrate L-glutamine concentration of 120 mmol/L, reaction temperature of 45℃, and L-theanine yield of 89.1%. The yield of L-theanine is 2.61 times higher than that obtained without ultrasound. Ultrasound can significantly promote the synthesis of L-theanine by γ-glutamyltranspeptidase.