期刊
CATALYSIS TODAY
卷 366, 期 -, 页码 185-194出版社
ELSEVIER
DOI: 10.1016/j.cattod.2020.04.002
关键词
Biocatalysis; Phenylalanine ammonia-lyase; Yeast; Pseudozyma antarctica; Unnatural amino acids
资金
- European Social Fund [P37_273, 103413, POCU/360/6/13/123886]
- National Authority for Scientific Research and Innovation (ANCSI)
- European Regional Development Fund, Competitiveness Operational Program 2014-2020 (POC)
- Swiss National Science Foundation (SNSF) [IZ11Z0_166543]
- Higher Education Excellence Program of the Ministry of Human Capacities (Budapest, Hungary) in the frame of Biotechnology research area of Budapest University of Technology and Economics (BME FIKP-BIO)
- National Research, Development and Innovation Fund of Hungary (Budapest, Hungary) [SNN-125637]
- CELSA (BME-KU Leuven)
- Swiss National Science Foundation (SNF) [IZ11Z0_166543] Funding Source: Swiss National Science Foundation (SNF)
A novel phenylalanine ammonia-lyase (PzaPAL) was identified from the psychrophilic yeast Pseudozyma antarctica, showing significantly different substrate binding pocket compared to known PALs. Recombinant PzaPAL exhibited higher turnover numbers than PcPAL and demonstrated enhanced enantiotope selectivity in both kinetic resolutions and ammonia addition reactions.
A novel phenylalanine ammonia-lyase of the psychrophilic yeast Pseudozyma antarctica (PzaPAL) was identified by screening microbial genomes against known PAL sequences. PzaPAL has a significantly different substrate binding pocket with an extended loop (26 aa long) connected to the aromatic ring binding region of the active site as compared to the known PALs from eukaryotes. The general properties of recombinant PzaPAL expressed in E. coli were characterized including kinetic features of this novel PAL with L-phenylalanine (S) -1a and further racemic substituted phenylalanines rac-1b-g,k. In most cases, PzaPAL revealed significantly higher turnover numbers than the PAL from Petroselinum crispum (PcPAL). Finally, the biocatalytic performance of PzaPAL and PcPAL was compared in the kinetic resolutions of racemic phenylalanine derivatives (rac-1a-s) by enzymatic ammonia elimination and also in the enantiotope selective ammonia addition reactions to cinnamic acid derivatives (2a-s). The enantiotope selectivity of PzaPAL with o-, m-, p-fluoro-, o-, p-chloro-and o-, m-bromo-substituted cinnamic acids proved to be higher than that of PcPAL.
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