Effects of salts on the conformation and catalytic properties of d-amino acid aminotransferase.
The effects of salts on the biochemical properties of D-amino acid aminotransferase from Bacillus sp. YM-1 have been studied to elucidate both the inhibitory effects of salts on the activity and the protective effects of salts on the substrate-induced inactivation. The results from UV-visible spectroscopy studies on the reaction of the enzyme with D-serine revealed that salt significantly reduced the rate of the formation of the quinonoid intermediate and its accumulation. The kinetic and spectroscopy studies of the reaction with alpha-[(2)H]-DL-serine in different concentrations of NaCl provided evidence that the rate-limiting step was changed from the deprotonation of the external aldimine to another step(s), presumably to the hydrolysis of the ketimine. Gel filtration chromatography data in the presence of NaCl showed that the enzyme volume was reduced sharply with the increasing NaCl concentration, up to 100 mM. An additional increase of the NaCl concentration did not affect the elution volume, which suggests that the enzyme has a limited number of salt-binding groups. These results provide detailed mechanistic evidence for the way salts inhibit the catalytic activity of Damino acid aminotransferase
Ro
Hyeon-Su
HS
Biomolecular Process Engineering Laboratory, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea. rohyeon@mail.kribb.re.kr
D000410:Alanine Transaminase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D001407:Bacillus / Q000201:enzymology*; D002384:Catalysis / Q000187:drug effects*; D002850:Chromatography, Gel; D002942:Circular Dichroism; D051263:D-Alanine Transaminase; D004735:Energy Transfer; D004926:Escherichia coli / Q000235:genetics; D007700:Kinetics; D011487:Protein Conformation / Q000187:drug effects*; D012694:Serine / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology*; D013057:Spectrum Analysis; D013237:Stereoisomerism
D012965:Sodium Chloride; D012694:Serine; D000410:Alanine Transaminase; D051263:D-Alanine Transaminase