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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Acknowledgments
This work is financially supported by the National High Technology Research and Development Program of China (863 Program, 2011AA100905), the Program for New Century Excellent Talents in University (NCET-10-0461), the Key project of Chinese Ministry of Education (311023), the Priority Academic Program Development of Jiangsu Higher Education Institutions, the 111 Project (No. 111-2-06).
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Zhu, L., Cui, W., Fang, Y. et al. Cloning, expression and characterization of phenylalanine ammonia-lyase from Rhodotorula glutinis . Biotechnol Lett 35, 751–756 (2013). https://doi.org/10.1007/s10529-013-1140-7
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DOI: https://doi.org/10.1007/s10529-013-1140-7