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Novel Salt-Tolerant Leucine Dehydrogenase from Marine Pseudoalteromonas rubra DSM 6842

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Abstract

This study reported the cloning, expression, and characterization of a new salt-tolerant leucine dehydrogenase (PrLeuDH) from Pseudoalteromonas rubra DSM 6842. A codon-optimized 1038 bp gene encoding PrLeuDH was successfully expressed on pET-22b( +) in E. coli BL21(DE3). The purified recombinant PrLeuDH showed a single band of about 38.7 kDa on SDS-PAGE. It exhibited the maximum activity at 40 °C and pH 10.5, while kept high activities in the range of 25–45 °C and pH 9.5–12. The Km value and turnover number kcat for leucine of PrLeuDH were 2.23 ± 0.12 mM and 35.39 ± 0.05 s−1, respectively, resulting in a catalytic efficiency kcat/Km of 15.87 s−1/mM. Importantly, PrLeuDH remained 92.1 ± 2.67% active in the presence of 4.0 M NaCl. The study provides the first in-depth understanding of LeuDH from marine Pseudoalteromonas rubra, meanwhile the unique properties of high activity at low temperature and high salt tolerance make it a promising biocatalyst for the synthesis of non-protein amino acids and α-ketoacids under special conditions in pharmaceutical industry.

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Acknowledgements

This work was supported by the Guangxi science and technology base and talent special project (No. AD211064) and the Beihai "13th five year plan" marine economic innovation and development demonstration project (No. Bhsfs010-4).

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  1. College of Light Industry and Food Engineering, Guangxi University, 100 Daxue East Road, Nanning, 530004, People’s Republic of China

    Rui Chen, Yu-Ting Liao, Tian-Tian Gao, Yan-Mei Zhang, Liang-Hua Lu & Cheng-Hua Wang

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Chen, R., Liao, YT., Gao, TT. et al. Novel Salt-Tolerant Leucine Dehydrogenase from Marine Pseudoalteromonas rubra DSM 6842. Mol Biotechnol 64, 1270–1278 (2022). https://doi.org/10.1007/s12033-022-00505-0

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