CRYSTALLINE ALCOHOL DEHYDROGENASES FROM THE MESOPHILIC BACTERIUM CLOSTRIDIUM-BEIJERINCKII AND THE THERMOPHILIC BACTERIUM THERMOANAEROBIUM-BROCKII - PREPARATION, CHARACTERIZATION AND MOLECULAR SYMMETRY

Two tetrameric NADP(+)-dependent bacterial secondary alcohol dehydroge nases have been crystallized in the apo- and the holoenzyme forms. Cry stals of the hole-enzyme from the mesophilic Clostridium beijerinckii (NCBAD) belong to space group P2(1)2(1)2(1) with unit-cell dimensions a = 90.5, b = 127.9, c = 151.4 Angstrom. Crystals of the ape-enzyme (C BAD) belong to the same space group with unit-cell dimensions a = 80.4 , b = 102.3, c = 193.5 Angstrom. Crystals of the hole-enzyme from the thermophilic Thermoanaerobium brockii (NTBAD) belong to space group P6 (1(5)) (a = b = 80.6, c = 400.7 Angstrom). Crystals of the ape-form of TBAD (point mutant G198D) belong to space group P2(1) with cell dimen sions a = 123.0, b = 84.8, c = 160.4 Angstrom beta = 99.5 degrees. Cry stals of CBAD, NCBAD and NTBAD contain one tetramer per asymmetric uni t. They diffract to 2.0 Angstrom resolution at liquid nitrogen tempera ture. Crystals of TBAD(G198D) have two tetramers per asymmetric unit a nd diffract to 2.7 Angstrom at 276 K. Self-rotation analysis shows tha t both enzymes are tetramers of 222 symmetry.