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Identification and Characterization of a Novel Soluble Pyridine Nucleotide Transhydrogenase from Streptomyces avermitilis

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Abstract

Soluble pyridine nucleotide transhydrogenase (STH) transfers hydride between NADH and NADPH to maintain redox balance. In the present study, the sth gene from Gram-positive bacterium Streptomyces avermitilis (SaSTH) was expressed in Escherichia coli, and the recombinant STH protein was purified to homogeneity. Activity assays indicated that SaSTH was able to catalyze transhydrogenase reactions by using NADH or NADPH as reductants and thio-NAD+ as an oxidant. The apparent Km value for NADPH (74.5 μM) was lower than that for NADH (104.0 μM) and the apparent kcat/Km for NADPH (2704.7 mM−1 s−1) was higher than that for NADH (1129.8 mM−1 s−1). SaSTH showed optimal activity at 25 °C and at a pH of 6.2. Heat-inactivation studies revealed that SaSTH remained stable below 55 °C and that approximately 50% activity was preserved at 57 °C for 20 min. Analyses also showed that SaSTH activity was inhibited by divalent ions, particularly Co2+, Ni2+, and Zn2+. In addition, the transhydrogenase activity of SaSTH was inhibited by ATP and strongly stimulated by ADP and AMP. In summary, we characterized a recombinant enzyme exhibiting STH activity from Gram-positive bacteria for the first time. Our findings provide new options for cofactor engineering and industrial biocatalytic processes.

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Data Availability

The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

This research was supported by the National Natural Science Foundation of China (Reference: 32071270), The Major Science and Technology Projects in Anhui Province (Reference: 202003a06020009), and Anhui Provincial Key Laboratory of the Conservation and Exploitation of Biological Resources.

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  1. Zhengyu Cao and Jie Liu have contributed equally to this research.

Authors and Affiliations

  1. Anhui Provincial Key Laboratory of Molecular Enzymology and Mechanism of Major Diseases and Key Laboratory of Biomedicine in Gene Diseases and Health of Anhui Higher Education Institutes, No.1 Beijing East Road, College of Life Sciences, Anhui Normal University, Anhui, 241000, China

    Zhengyu Cao, Jie Liu, Rui Meng, Peng Wang & Guoping Zhu

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Contributions

ZC, JL, and RM performed the experiments. ZC and GZ designed the project. ZC and JL analyzed the data. ZC, JL, PW, and GZ wrote the manuscript. All authors reviewed the manuscript and approved the final version for publication.

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Correspondence to Peng Wang or Guoping Zhu.

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Cao, Z., Liu, J., Meng, R. et al. Identification and Characterization of a Novel Soluble Pyridine Nucleotide Transhydrogenase from Streptomyces avermitilis. Curr Microbiol 79, 32 (2022). https://doi.org/10.1007/s00284-021-02727-y

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