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Cloning, expression and characterization of phenylalanine ammonia-lyase from Rhodotorula glutinis |
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| Authors: | Longbao Zhu Wenjing Cui Yueqin Fang Yi Liu Xinxing Gao Zhemin Zhou |
| Institution: | 1. Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu, China 3. School of Biochemical Engineering, Anhui Polytechnic University, 8 Zheshan Road, Wuhu, Anhui, China 2. School of Environmental and Civil Engineering, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu, China |
| 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 × 104 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. |
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