Screening and simple purification of alanine dehydrogenase in Bacillus strains.
Alanine dehydrogenase (EC 1.4.1.1), in the presence of NAD+, catalyzes the reversible deamination of L-alanine. Screening of alanine dehydrogenase in bacillus strains was carried out to develop its utilization as an industrial and analytical catalyst. Eight bacillus strains were used, including Bacillus megaterium LA 199 which abundantly produces enzymes. Alanine dehydrogenase was purified simply from Bacillus megaterium LA 199 by heat treatment at pH 5.4, followed by DEAE-Sepharose CL-6B and Sepharose CL-2B chromotography. The enzyme consisted of six subunits with an identical molecular mass of 42.5 kDa. The Km were 1.17 x 10(-2) mM for NADH and 5.12 x 10(-2) mM for pyruvate.
Baysal
Senay Hamarat
SH
Department of Biochemistry, a Ege University, Faculty of Science, Izmir, Turkey. hamarat@sci.ege.edu.tr
Yasa
Ihsan
I
Uslan
Arif H
AH
D050788:Alanine Dehydrogenase; D000594:Amino Acid Oxidoreductases / Q000302:isolation & purification* / Q000378:metabolism; D001407:Bacillus / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D009243:NAD / Q000378:metabolism
D009243:NAD; D000594:Amino Acid Oxidoreductases; D050788:Alanine Dehydrogenase