Almando Geraldi, .- and Ni’matuzahroh and Fatimah, .- and Chang-Hao Cui, .- and Thi Thuy Nguyen, .- and Sun Changkim, .- (2020) Enzymatic biotransformation of ginsenoside Rb1 by recombinant β-glucosidase of bacterial isolates from Indonesia. Biocatalysis and Agricultural Biotechnology, 23 (101449). pp. 1-8. ISSN 101449

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Abstract

In this investigation, Bacillus licheniformis γ-glutamyltranspeptidase (BlGGT) together with its two transpeptidase-specialized mutants (N450D and N450Q) were employed for the biocatalytic synthesis of L-theanine. A high-pressure liquid chromatographic method coupled with ultraviolet detection (HPLC/UV) was used to quantify L-theanine content in the reaction mixture. Through a series of experiments, the perfect conditions for the biocatalytic synthesis were found to be an operational pH of 10.5, a reaction time of 4 h, L-glutamine and ethanolamine in a 1:2.4 M ratio, and enzymes at a working concentration of 25 μg/mL. In a batch process for N450D-mediated synthesis of L-theanine at 37 °C, a conversion rate of ~94% was achieved from 250 mM l-glutamine and 600 mM ethylamine. Successful synthesis of the desired product was further verified by mass spectrometry analysis. Conclusively, the experimental results suggest a high potential application in the production of L-theanine by BlGGT-mediated biocatalysis.

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