Molecular characterization of a phenylalanine ammonia-lyase gene (BoPAL1) from Bambusa oldhamii
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- 作者：Lu-Sheng Hsieh ; Yi-Lin Hsieh ; Chuan-Shan Yeh ; Chieh-Yang Cheng ; Chien-Chih Yang and Ping-Du Lee
- 关键词：Phenylalanine ammonia ; lyase (PAL) ; Bamboo (Bambusa oldhamii) ; Molecular cloning ; TAIL ; PCR ; Gene expression
- 刊名：Molecular Biology Reports
- 出版年：2011
- 出版时间：January 2011
- 年：2011
- 卷：38
- 期：1
- 页码：283-290
- 全文大小：514.7 KB
文摘
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¡ãC 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.