SCREENING AND SIMPLE PURIFICATION OF ALANINE DEHYDROGENASE IN BACILLUS STRAINS
Articles > Article ยท 2002
Authors: Ihsan Yasa, Arif H. Uslan1, Senay Hamarat Baysal
Preparative Biochemistry & Biotechnology, International, WoS, 2002, vol 32, pp 277-285
DOI: 10.1081/pb-120013474
Summary
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.