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Molecular Cloning and Expression Analysis of the Gene Encoding Proline Dehydrogenase from Jatropha curcas L

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Abstract

Proline dehydrogenase (ProDH) (EC 1.5.99.8) is a key enzyme in the catabolism of proline. The enzyme JcProDH and its complementary DNA (cDNA) were isolated from Jatropha curcas L., an important woody oil plant used as a raw material for biodiesels. It has been classified as a member of the Pro_dh superfamily based on multiple sequence alignment, phylogenetic characterization, and its role in proline catabolism. Its cDNA is 1674 bp in length with a complete open reading frame of 1485 bp, which encodes a polypeptide chain of 494 amino acids with a predicted molecular mass of 54 kD and a pI of 8.27. Phylogenetic analysis indicated that JcProDH showed high similarity with ProDH from other plants. Reverse transcription PCR (RT-PCR) analysis revealed that JcProDH was especially abundant in the seeds and flowers but scarcely present in the stems, roots, and leaves. In addition, the expression of JcProDH increased in leaves experiencing environmental stress such as cold (5 °C), heat (42 °C), salt (300 mM), and drought (30 % PEG6000). The JcProDH protein was successfully expressed in the yeast strain INVSc1 and showed high enzyme activity in proline catabolism. This result confirmed that the JcProDH gene negatively participated in the stress response.

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Acknowledgments

This work was supported by several grants from the National Foundations of Natural Sciences, China (No. 31260064 to MG, No. 31260169 to SLY), and the Education Bureau of Yunnan Province (No. ND2010004 to MG).

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Authors and Affiliations

  1. School of Life Sciences, Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Key Laboratory of Biomass Energy and Environmental Biotechnology of Yunnan Province, Yunnan Normal University, Kunming, 650500, Yunnan, People’s Republic of China

    Haibo Wang, Pingxing Ao, Shuanglong Yang, Zhurong Zou, Shasha Wang & Ming Gong

  2. College of Biological Resources and Environmental Science, Qujing Normal University, Qujing, 655011, Yunnan, People’s Republic of China

    Haibo Wang

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  1. Haibo Wang
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Correspondence to Ming Gong.

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H. Wang and P. Ao have contributed equally to this work.

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Wang, H., Ao, P., Yang, S. et al. Molecular Cloning and Expression Analysis of the Gene Encoding Proline Dehydrogenase from Jatropha curcas L. Appl Biochem Biotechnol 175, 2413–2426 (2015). https://doi.org/10.1007/s12010-014-1441-x

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