title: Cloning and expression of a phenylalanine ammonia-lyase gene (BoPAL2) from Bambusa oldhamii in Escherichia coli and Pichia pastoris.
abstract: Phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) is the first committed enzyme of phenylpropanoid pathway. A PAL gene, designated as BoPAL2, was cloned from a Bambusa oldhamii cDNA library. The open reading frame of BoPAL2 was 2142bp in size encoding a 713-amino acid polypeptide. BoPAL2 was heterologous expressed in Escherichia coli and Pichia pastoris. The recombinant proteins were exhibited PAL and tyrosine ammonia-lyase activities. The recombinant BoPAL2 had a subunit mass of 80kDa and existed as a homotetramer. The optimum temperature and pH of BoPAL2 were 50-60 degrees C and 8.5-9.0, respectively. The K(m) and k(cat) values of BoPAL2 expressed in E. coli were 250microM and 10.12s(-1). The K(m) and k(cat) values of BoPAL2 expressed in P. pastoris were 331microM and 16.04s(-1). The recombinant proteins had similar biochemical properties and kinetic parameters with PALs reported in other plants.
authors.affiliation: Institute of Microbiology and Biochemistry, Department of Biochemical Science and Technology, College of Life Science, National Taiwan University, Taipei, Taiwan.
authors.forename: Lu-Sheng
authors.initials: LS
authors.lastname: Hsieh
authors.forename: Chuan-Shan
authors.initials: CS
authors.lastname: Yeh
authors.forename: Hung-Chi
authors.initials: HC
authors.lastname: Pan
authors.forename: Chieh-Yang
authors.initials: CY
authors.lastname: Cheng
authors.forename: Chien-Chih
authors.initials: CC
authors.lastname: Yang
authors.forename: Ping-Du
authors.initials: PD
authors.lastname: Lee
mesh_terms: D000642:Ammonia-Lyases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D031723:Bambusa / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015723:Gene Library; D016366:Open Reading Frames; D010650:Phenylalanine Ammonia-Lyase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D000642:Ammonia-Lyases; C023208:L-tyrosine ammonia-lyase; D010650:Phenylalanine Ammonia-Lyase