Journal
FOOD BIOTECHNOLOGY
Volume 32, Issue 2, Pages 130-147Publisher
TAYLOR & FRANCIS INC
DOI: 10.1080/08905436.2018.1444636
Keywords
gamma-glutamyl-L-leucine; gamma-Glutamyltranspeptidase; biocatalytic synthesis; bacillus licheniformis; thin-layer charomatography
Funding
- Ministry of Science and Technology of Taiwan [MOST-103-2313-B-415-008-MY3]
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Several -glutamyl compounds are known to have favorable features for a variety of biotechnological and pharmaceutical applications. However, so far only a limited number of -glutamyl compounds have been synthesized through -glutamyltranspeptidase (GGT). With the help of thin layer chromatography and computer-assisted image analysis techniques, we performed an enzymatic method involving Bacillus licheniformis GGT (BlGGT) for the synthesis of -glutamyl-L-leucine (-Glu-Leu). The optimum conditions for a biocatalytic synthesis of -Glu-Leu were determined to be 200mM glutamine, 200mM leucine, 50mM Tris-HCl buffer (pH 9.0), and BlGGT at a working concentration of 2.0U/mL. Upon a 5-h incubation of the reaction mixture at 50 degrees C, the product yield could reach approximately 52%. The preparative synthesis of -Glu-Leu by BlGGT was done under optimal conditions at millimol scale level. The reaction products were subjected to mass spectrometry/nuclear magnetic resonance analyses to ensure the successful synthesis of -Glu-Leu. Taken together, the present study demonstrates the high potentiality of BlGGT-catalyzed reaction in the synthesis of -glutamyl compounds.
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