Overexpression of salicylate hydroxylase and the crucial role of lys(163) as its NADH binding site

• Published in: Journal of biochemistry (Tokyo) Vol. 128; no. 2; p. 293
• Main Authors: Suzuki, K, Asao, E, Nakamura, Y, Nakamura, M, Ohnishi, K, Fukuda, S
• Format: Journal Article
• Language: English
• Published: England 01.08.2000
• Subjects: Amino Acid Substitution; Apoenzymes - metabolism; Binding Sites; Cell-Free System; Electrophoresis, Polyacrylamide Gel; Escherichia coli - metabolism; Hydrogen-Ion Concentration; Inclusion Bodies - metabolism; Kinetics; Lysine - genetics; Mixed Function Oxygenases - chemistry; Mixed Function Oxygenases - genetics; Mixed Function Oxygenases - metabolism; Molecular Sequence Data; Mutagenesis, Site-Directed; NAD - metabolism; Pseudomonas putida - chemistry

Expression systems for the sal gene encoding salicylate hydroxylase from Pseudomonas putida S-1 were examined and some constructs were expressed in these systems. By cultivation of Escherichia coli BL21 (DE3)/pSAH8 in LB medium at 37 degrees C with isopropyl-b-D-thiogalactopyranoside as the inducer,...