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

作者：Hsieh Lu Sheng; Hsieh Yi Lin; Yeh Chuan Shan; Cheng Chieh Yang; Yang Chien Chih; Lee Ping Du

来源：Molecular Biology Reports, 2011, 38(1): 283-290.

DOI：10.1007/s11033-010-0106-2

• Phenylalanine ammonia-lyase (PAL)
• Bamboo (Bambusa oldhamii)
• Molecular cloning
• TAIL-PCR
• Gene expression

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 50A degrees 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.

• 出版日期2011-1