Identification, Heterologous Expression and Characterization of a Transaminase from Rhizobium sp.

Abstract

In present study, we have mined a transaminase (TA) from Rhizobium sp. from the pool of fully sequenced genomes by using an ω-TA sequence from Vibrio fluvialis JS17 as a template in a BLASTP search. The protein sequence of the TA from Rhizobium sp. exhibits 53% sequence identity to that from V. fluvialis. The TA with S-selectivity showed close evolutionary relationship with a pyruvate transaminase from Alcaligenes denitrificans Y2k-2 and an S-selective aminotransferase from Sphaerobacter thermophilus. The gene of the ω-TA was inserted into pET-28a and functionally expressed in E. coli BL21. Results showed that the recombinant ω-TA has a specific activity of 7.46 U/mg at pH 8.0, 30 ℃. The substrate specificity test found the ω-TA presented significant reactivity toward aromatic amino donors and amino acceptors containing aldehydes. More importantly, the ω-TA also exhibited a good affinity towards some cyclic substrates. The homology model of the ω-TA was built by Discovery Studio and docking was performed to describe the relative activity towards some substrates.

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Acknowledgments

The authors gratefully acknowledge the financial support from NSFC (21878155), Provincial Key R&D Plan of Jiangsu (BE2017703), PAPD, Qing Lan Project of Jiangsu Universities, Six Talent Peaks Project in Jiangsu Province, and the Jiangsu Synergetic Innovation Center for Advanced Bio-Manufacture.

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K.X.T. conducted most of the experiments. Y.L. and H.H.J. designed and supervised the project. Y.F.Y performed activity assay and tested the related reactivity of substrates. All the authors discussed the design and results, commented on the manuscript, and approved the manuscript.

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Correspondence to Honghua Jia or Yan Li.

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Tang, K., Yi, Y., Gao, Z. et al. Identification, Heterologous Expression and Characterization of a Transaminase from Rhizobium sp.. Catal Lett 150, 2415–2426 (2020). https://doi.org/10.1007/s10562-020-03121-2

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Keywords

  • Transaminase
  • Rhizobium sp.
  • Characterization
  • Substrate specificity