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KMID : 1094720210260030408
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Biotechnology and Bioprocess Engineering
2021 Volume.26 No. 3 p.408 ~ p.418
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Exploiting the Reversibility of GTBP1 Catalyzed One-pot Reactions for the Synergistical Synthesis of Ponasterone A and Phenolic Glycosides
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Li Yuqiang
Li Bingfeng
Chen Tianyi
Wu Bin
Chu Jianlin
He Bingfang
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Abstract
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In the process of glycosyltransferase-catalyzed transglycosylation, the synthesis of uridine diphosphate sugars is generally thermodynamically disfavored. This study found that GTBP1 from Bacillus pumilus BF1 displayed the potential to reversely synthesize uridine diphosphate glucose (UDPG, UDP-glucose), which catalyzed the deglycosylation of ponasteroside A and the transglycosylation of phenolic compounds in one-pot reaction. Sequence alignment and phylogenetic tree analysis of GTBP1 and other GTs with reversible glycosylation ability were also implemented. Using solvent engineering strategy, the reaction time in water-organic biphasic was shortened and the conversion was improved. The final conversion of ponasterone A was reached 93.4%, and the final yield of ferulic acid glycoside was 92.7%. Using the fed-batch technology with the coupled reaction, the cumulative product of ponasterone A was about 1.97 g/L, and the level of produced ferulic acid glycoside was about 1.52 g/L. The substrate specificity of the GTBP1 was also confirmed. It implies the diversity of GTBP1’s ability to construct UDP cycles.
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KEYWORD
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microbial glycosyltransferases, catalytic reversibility, UDPG synthesis, solvent engineering, deglycosylation and transglycosylation
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