title: Purification and characterization of a novel (R)-imine reductase from Streptomyces sp. GF3587.
abstract: The (R)-imine reductase (RIR) of Streptomyces sp. GF3587 was purified and characterized. It was found to be a NADPH-dependent enzyme, and was found to be a homodimer consisting of 32 kDa subunits. Enzymatic reduction of 10 mM 2-methyl-1-pyrroline (2-MPN) resulted in the formation of 9.8 mM (R)-2-methylpyrrolidine ((R)-2-MP) with 99% e.e. The enzyme showed not only reduction activity for 2-MPN at neutral pH (6.5-8.0), but also oxidation activity for (R)-2-MP under alkaline pH (10-11.5) conditions. It appeared to be a sulfhydryl enzyme based on the sensitivity to sulfhydryl specific inhibitors. It was very specific to 2-MPN as substrate.
authors: Department of Biomolecular Science, Faculty of Engineering, Gifu University. mitukura@gifu-u.ac.jp
authors: Koichi
authors: K
authors: Mitsukura
authors: Mai
authors: M
authors: Suzuki
authors: Sho
authors: S
authors: Shinoda
authors: Tatsuya
authors: T
authors: Kuramoto
authors: Toyokazu
authors: T
authors: Yoshida
authors: Toru
authors: T
authors: Nagasawa
mesh_terms: D002850:Chromatography, Gel; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007097:Imines / Q000378:metabolism*; D007700:Kinetics; D008745:Methylation; D008970:Molecular Weight; D009249:NADP / Q000378:metabolism; D010088:Oxidoreductases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D021122:Protein Subunits / Q000737:chemistry*; D011758:Pyrroles / Q000378:metabolism*; D011759:Pyrrolidines / Q000378:metabolism*; D013302:Streptomyces / Q000737:chemistry / Q000201:enzymology*; D013379:Substrate Specificity; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D020245:p-Chloromercuribenzoic Acid / Q000494:pharmacology
chemical_list: C520125:2-methylpyrrolidine; D007097:Imines; D021122:Protein Subunits; D011758:Pyrroles; D011759:Pyrrolidines; D013439:Sulfhydryl Reagents; C013231:pyrroline; D009249:NADP; D020245:p-Chloromercuribenzoic Acid; D010088:Oxidoreductases