title: Cloning, expression and characterization of phenylalanine ammonia-lyase from Rhodotorula glutinis.
abstract: The industrial-scale production of phenylalanine ammonia-lyase (PAL) mainly uses strains of Rhodotorula. However, the PAL gene from Rhodotorula has not been cloned. Here, the full-length gene of PAL from Rhodotorula glutinis was isolated. It was 2,121 bp, encoding a polypeptide with 706 amino acids and a calculated MW of 75.5 kDa. Though R. glutinis is an anamorph of Rhodosporium toruloides, the amino acid sequences of PALs them are not the same (about 74 % identity). PAL was expressed in E. coli and characterized. Its specific activity was 4.2 U mg(-1) and the k cat/K m was 1.9 × 10(4) mM(-1) s(-1), exhibiting the highest catalytic ability among the reported PALs. The genetic and biochemical information reported here should facilitate future application in industry.
authors: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu, China.
authors: Longbao
authors: L
authors: Zhu
authors: Wenjing
authors: W
authors: Cui
authors: Yueqin
authors: Y
authors: Fang
authors: Yi
authors: Y
authors: Liu
authors: Xinxing
authors: X
authors: Gao
authors: Zhemin
authors: Z
authors: Zhou
mesh_terms: D003001:Cloning, Molecular; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D019216:Metals, Heavy; D010650:Phenylalanine Ammonia-Lyase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012248:Rhodotorula / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity; D013501:Surface-Active Agents; D013696:Temperature
chemical_list: D005656:Fungal Proteins; D019216:Metals, Heavy; D011994:Recombinant Proteins; D013501:Surface-Active Agents; D010650:Phenylalanine Ammonia-Lyase