Engineering of microheterogeneity‐resistant p‐hydroxybenzoate hydroxylase from Pseudomonas fluorescens

FEBS Letters

Mattevi, Andrea²  Obmolova, Galina²  Kalk, Kor H.²  Westphal, Adrie H.¹  de Kok, Arie¹  Hol, Wim G.J.²  van Berkel, Willem J.H.¹  Eschrich, Klaus¹ 

[1] Department of Biochemistry, Agricultural University, Dreijenlaan 3, 6703 HA Wageningen, The Netherlands;BIOSON Research Institute, Department of Chemistry, University ofGroningen, Nijenborgh 16, 9747 AG Groningen, The Netherlands

关键词: p-Hydroxybenzoate hydroxylase;    Protein-engineering;    Site-directed mutagenesis;    Thiol-reactivity;    Crystallization;   

DOI  :  10.1016/0014-5793(90)80843-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

By site-directed mutagenesis, Cys-116 was converted to Ser-116 in p-hydroxybenzoate hydroxylase (EC 1.14.13.2) from Pseudomonas fluorescens. In contrast to wild-type enzyme, the C116S mutant is no longer susceptible to oxidation by hydrogen peroxide and shows no reactivity towards 5,5'-dithiobis(2-nitrobenzoate). Crystals of the C116S mutant are isomorphous with the crystal form of wild-type enzyme. A difference electron density confirms the mutation made.

【 授权许可】

Unknown