Alteration in relative activities of phenylalanine dehydrogenase towards different substrates by site‐directed mutagenesis

FEBS Letters

Baker, Patrick J.³  Rice, David W.³  Asano, Yasuhisa¹  Linda Britton, K.³  Engel, Paul C.²  Seah, Stephen Y.K.² 

[1] Biotechnology Research Center, Toyama Prefectural University, 5180 Kurokawa, Kosugi, Toyama 939-03, Japan;Department of Biochemistry, University College Dublin, Merville House, Belield, Dublin 4, Ireland;The Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, University of Sheffield, PO Box 594, Sheffield S10 2UH, UK

关键词: Phenylalanine dehydrogenase;    Mutagenesis;    Substrate specificity;    Amino acids;   

DOI  :  10.1016/0014-5793(95)00804-I

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Glycine-124 and leucine-307 of phenylalanine dehydrogenase from Bacillus sphaericus were altered by site-specific mutagenesis to the corresponding residues in leucine dehydrogenase: alanine and valine, respectively. These two residues have previously been implicated from molecular modelling as important in determining the substrate discrimination of the two enzymes. Single and double mutants displayed lower activities towards l-phenylalanine and enhanced activity towards almost all aliphatic amino acid substrates tested compared to the wild-type, thus confirming the predictions made from molecular modelling.

【 授权许可】

Unknown