Abstract
Glycosyltransferase 1 from Bacillus cereus (BcGT1) catalyzes the transfer of a glucosyl moiety from uridine diphosphate glucose (UDP-glucose) to various acceptors; it was expressed and characterized. The specificity of acceptors was found to be broad: more than 20 compounds classified into O-, S-, and N-linkage glucosides can be prepared with BcGT1 catalysis. Based on this work, we conclude that the corresponding acceptors of these compounds must possess the following features: (1) the acceptors must contain at least one aromatic or fused-aromatic or heteroaromatic ring; (2) the reactive hydroxyl or sulfhydryl or amino group can attach either on the aromatic ring or on its aliphatic side chain; and (3) the acceptors can be a primary, secondary, or even a tertiary amine. Four representative acceptors—fluorescein methyl ester, 17-β-estradiol, 7-mercapto-4-methylcoumarin, and 6-benzylaminopurine—were chosen as a candidate acceptor for O-, S-, and N-glucosidation, respectively. These enzymatic products were purified and the structures were confirmed with mass and NMR spectra. As all isolated glucosides are β-anomers, BcGT1 is confirmed to be an inverting enzyme. This study not only demonstrates the substrate promiscuity of BcGT1 but also showed the great application prospect of this enzyme in bioconversion of valuable bioactive molecules.
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Acknowledgments
We thank Ms. Chia-Yu Lu, Ms. Ko-Jen Su for enzyme preparation, Dr. Hsing-Pang Hsieh (National Health Research Institutes, Taiwan) for providing BPR2P0064S0, Mr. Chang-Lin Yang for isolating 7-mercapto-4-methylcoumarin 7-S-glucoside, Dr. Kuang-Po Chen, Ms. Chloe Chang and Dr. Li-Ching Shen for the NMR analysis, Ms. Yun-Ming Li (Mass Laboratory, Center for Advanced Instrumentation at National Chiao Tung University, Taiwan) for mass analysis, and Dr. Yu-Kuo Wang (Department of Biological Science and Technology, National Chiao Tung University, Taiwan) for MALDI-TOF-MS peptide-mapping analysis.
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This study was funded by the Ministry of Science and Technology in Taiwan (101-2113-M-009-012-MY3).
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Chiu, HH., Shen, MY., Liu, YT. et al. Diversity of sugar acceptor of glycosyltransferase 1 from Bacillus cereus and its application for glucoside synthesis. Appl Microbiol Biotechnol 100, 4459–4471 (2016). https://doi.org/10.1007/s00253-015-7270-1
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DOI: https://doi.org/10.1007/s00253-015-7270-1