title: Malate dehydrogenase from the mesophile Chlorobium vibrioforme and from the mild thermophile Chlorobium tepidum: molecular cloning, construction of a hybrid, and expression in Escherichia coli.
abstract: The genes (mdh) encoding malate dehydrogenase (MDH) from the mesophile Chlorobium vibrioforme and the moderate thermophile C. tepidum were cloned and sequenced, and the complete amino acid sequences were deduced. When the region upstream of mdh was analyzed, a sequence with high homology to an operon encoding ribosomal proteins from Escherichia coli was found. Each mdh gene consists of a 930-bp open reading frame and encodes 310 amino acid residues, corresponding to a subunit weight of 33,200 Da for the dimeric enzyme. The amino acid sequence identity of the two MDHs is 86%. Homology searches using the primary structures of the two MDHs revealed significant sequence similarity to lactate dehydrogenases. A hybrid mdh was constructed from the 3' part of mdh from C. tepidum and the 5' part of mdh from C. vibrioforme. The thermostabilities of the hybrid enzyme and of MDH from C. vibrioforme and C. tepidum were compared.
authors.affiliation: Department of Biology, University of Oslo, Norway.
authors.forename: K
authors.initials: K
authors.lastname: Naterstad
authors.forename: V
authors.initials: V
authors.lastname: Lauvrak
authors.forename: R
authors.initials: R
authors.lastname: Sirevåg
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D019414:Chlorobi / Q000201:enzymology*; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000378:metabolism; D015870:Gene Expression; D006361:Heating; D008291:Malate Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid
chemical_list: D004269:DNA, Bacterial; D008291:Malate Dehydrogenase