[Purification and characterization of carbonyl enantioselective reductase from Morganella morganii J-8].
The purification and the characteristics of an enzyme from Morganella morganii J-8, which could produce d-pseudoephedrine from 1-phenyl-2-methylamine-acetone, were performed in this study. In this research, first, cells were disrupted by ultrasonic treatment at 4 degrees C. The carbonyl enantioselective reductase was purified with a combination of ammonium precipitation, Phenyl Superose hydrophobic chromatography, DEAE anion exchange, and native polyacrylamide gel electrophoresis. The molecular mass of the purified enzyme subunit was estimated to be 42.5kD on sodium dodecyl sulfate-polyacrylamide electrophoresis (SDS-PAGE). The native molecular mass of the enzyme that was analyzed by high-performance liquid chromatography was found out to be 84.1 kD, which indicated that the enzyme was a dimmer. The purified enzyme was analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and the result showed that the purified enzyme had high homology with leucine dehydrogenase.
Zhang
Peng-Hu
PH
Laboratory of Biomass Resources, Southern Yangtze University, Wuxi 214036, China.
Zhang
Liang
L
Lu
Yan
Y
Shi
Gui-Yang
GY
D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D055162:Biocatalysis / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D050803:Leucine Dehydrogenase / Q000378:metabolism; D019216:Metals, Heavy / Q000494:pharmacology; D008970:Molecular Weight; D020613:Morganella morganii / Q000201:enzymology* / Q000378:metabolism; D010088:Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D054199:Pseudoephedrine / Q000737:chemistry / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013237:Stereoisomerism; D013696:Temperature
D001426:Bacterial Proteins; D019216:Metals, Heavy; D054199:Pseudoephedrine; D010088:Oxidoreductases; D050803:Leucine Dehydrogenase