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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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Abstract

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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Acknowledgments

This work was supported by the National Key Research and Development Program of China (2018YFA0902000), National Natural Science Foundation of China (81673321, 21776135, 21506099), Natural Science Foundation of Jiangsu province (Grant No. BK20181379). We also thank the Jiangsu Synergetic Innovation Center for Advanced Bio-Manufacture (NO. XTE1854 and NO. XTC1812). We are grateful to the High Performance Computing Center of Nanjing Tech University for supporting the computational resources. The authors declare no conflict of interest.

Neither ethical approval nor informed consent was required for this study.

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Authors and Affiliations

  1. College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing, 211800, China

    Yuqiang Li, Tianyi Chen & Bin Wu

  2. School of Biology and Environment, Nanjing Polytechnic Institute, Nanjing, 210048, China

    Bingfeng Li

  3. School of Pharmaceutical Sciences, Nanjing Tech University, Nanjing, 211800, China

    Jianlin Chu & Bingfang He

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  1. Yuqiang Li
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Correspondence to Jianlin Chu or Bingfang He.

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Li, Y., Li, B., Chen, T. et al. Exploiting the Reversibility of GTBP1 Catalyzed One-pot Reactions for the Synergistical Synthesis of Ponasterone A and Phenolic Glycosides. Biotechnol Bioproc E 26, 408–418 (2021). https://doi.org/10.1007/s12257-020-0135-1

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