Abstract
Objective
To characterize a novel xanthine dehydrogenase (XDH) from Acinetobacter baumannii by recombinant expression in Escherichia coli and to assess its potential for industrial applications.
Results
The XDH gene cluster was cloned from A. baumannii CICC 10254, expressed heterologously in E. coli and purified to homogeneity. The purified recombinant XDH consisted of two subunits with the respective molecular weights of 87 kDa and 56 kDa according to SDS-PAGE. XDH catalysis was optimum at pH 8.5 and 40–45 °C, was stable under alkaline conditions (pH 7–11) and the half-inactivation temperature was 60 °C. The K m, turnover number and catalytic efficiency for xanthine were 25 μM, 69 s−1 and 2.7 μM−1 s−1, respectively, which is an improvement over XDHs characterized previously. A. baumannii XDH is less than 50 % identical to previously identified XDH orthologs from other species, and is the first from the Acinetobacter genus to be characterized.
Conclusion
The novel A. baumannii enzyme was found to be among the most active, thermostable and alkaline-tolerant XDH enzymes reported to date and has potential for use in industrial applications.
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Acknowledgments
This work was supported by grants from the National Natural Science Foundation of China (Grant Number: 21406132), and the China Postdoctoral Science Foundation (Grant Numbers: 2014M550743 and 2015T80094).
Supporting information
Supplementary Table 1 Comparison between Acinetobacter baumannii xanthine dehydrogenase (XDH) identified in this study and previously characterized enzymes.
Supplementary Table 2 Amino acid sequence identity (homology) of A. baumannii XDH identified in this study and other XDHs characterized previously.
Supplementary Table 3 Purification of recombinant A. baumannii XDH following heterologous expression in E. coli.
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Wang, CH., Zhao, TX., Li, M. et al. Characterization of a novel Acinetobacter baumannii xanthine dehydrogenase expressed in Escherichia coli . Biotechnol Lett 38, 337–344 (2016). https://doi.org/10.1007/s10529-015-1986-y
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DOI: https://doi.org/10.1007/s10529-015-1986-y