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Molecular characterization of a phenylalanine ammonia-lyase gene (BoPAL1) from Bambusa oldhamii

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Abstract

Phenylalanine ammonia-lyase is the first enzyme of general phenylpropanoid pathway. A PAL gene, designated as BoPAL1, was cloned from a Bambusa oldhamii cDNA library. The open reading frame of BoPAL1 was 2,139 bp in size and predicted to encode a 712-amino acid polypeptide. BoPAL1 was the first intronless PAL gene found in angiosperm plant. Several putative cis-acting elements such as P box, GT-1motif, and SOLIPs involved in light responsiveness were found in the 5′-flanking sequence of BoPAL1 which was obtained by TAIL-PCR method. Recombinant BoPAL1 protein expressed in Pichia pastoris was active. The optimum temperature and pH for BoPAL1 activity was 50°C and 9.0, respectively. The molecular mass of recombinant BoPAL1 was estimated as 323 kDa using gel filtration chromatography and the molecular mass of full-length BoPAL was about 80 kDa, indicating that BoPAL1 presents as a homotetramer. The K m and k cat values of BoPAL1 for L-Phe were 1.01 mM and 10.11 s−1, respectively. The recombinant protein had similar biochemical properties with PALs reported in other plants.

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Acknowledgements

This work was supported by grants from the National Science Council of the Republic of China (Taiwan). We thank Prof. Ai-Yu Wang (National Taiwan University) for providing the Bambusa oldhamii cDNA library.

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Authors and Affiliations

  1. Institute of Microbiology and Biochemistry, Department of Biochemical Science and Technology, College of Life Science, National Taiwan University, 1 Roosevelt Road Section 4, Taipei, 106, Taiwan, ROC

    Lu-Sheng Hsieh, Yi-Lin Hsieh, Chuan-Shan Yeh, Chieh-Yang Cheng, Chien-Chih Yang & Ping-Du Lee

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  1. Lu-Sheng Hsieh
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Correspondence to Chien-Chih Yang or Ping-Du Lee.

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Hsieh, LS., Hsieh, YL., Yeh, CS. et al. Molecular characterization of a phenylalanine ammonia-lyase gene (BoPAL1) from Bambusa oldhamii . Mol Biol Rep 38, 283–290 (2011). https://doi.org/10.1007/s11033-010-0106-2

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