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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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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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
- Rhizobium sp.
- Substrate specificity