[Purification and properties of alanine dehydrogenase from Streptomyces lincolnensis].
Alanine Dehydrogenase (L-Alanine: NAD+ oxidoreductase, deaminating, EC 1.4.1.1) was purified from Streptomyces lincolnensis through four steps: (NH4)2SO4 precipitation, DEAE-cellulose 52, Affi-Gel Blue and Sepharose 6B. Molecular weight of the enzyme was determined as 170,000 by gel filtration and concentration gradient PAGE. SDS-PAGE showed only one band of 42,500, demonstrating that ADH from Streptomyces lincolnensis was consisted of four identical subunits. The optimal pH for amination was 9.0, for deamination 9.5. The optimal temperature for both amination and deamination was 50 degrees C. The Km valuse for pyruvate, NH4+, NADH, L-Ala and NAD+ were 2.08 x 10(-4) mol/L, 2.00 x 10(-2) mol/L, 2.38 x 10(-5) mol/L, 1.43 x 10(-2) mol/L and 6.67 x 10(-5) mol/L, respectively.
Jin
Z
Z
Shanghai Institute of Plant Physiology, Chinese Academy of Sciences, Shanghai 200032.
Jiao
R
R
D050788:Alanine Dehydrogenase; D000586:Amination; D000594:Amino Acid Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D013302:Streptomyces / Q000201:enzymology*; D013696:Temperature
D000594:Amino Acid Oxidoreductases; D050788:Alanine Dehydrogenase