Purification and properties of formate dehydrogenase from Moraxella sp. strain C-1.
نویسندگان
چکیده
NAD+-dependent formate dehydrogenase was screened in various bacterial strains. Facultative methanol-utilizing bacteria isolated from soil samples, acclimated to a medium containing methanol and formate at pH 9.5, were classified as members of the genus Moraxella. From a crude extract of Moraxella sp. strain C-1, formate dehydrogenase was purified to homogeneity, as judged by disc gel electrophoresis. The enzyme has an isoelectric point of 3.9 and a molecular weight of approximately 98,000. The enzyme is composed of two identical subunits with molecular weights of about 48,000. The apparent Km values for sodium formate and NAD+ were calculated to be 13 mM and 0.068 mM, respectively.
منابع مشابه
Cloning, nucleotide sequencing, and expression in Escherichia coli of the gene for formate dehydrogenase of Paracoccus sp. 12-A, a formate-assimilating bacterium.
The gene for the NAD-dependent formate dehydrogenase (FDH) of Paracoccus sp. 12-A, a formate-assimilating bacterium, was cloned through screening of the genomic library with activity staining. The FDH gene included an open reading frame of 1,200 base pairs, and encoded a protein of 43,757 Da, which had high amino acid sequence identity with known FDHs, in particular, with bacterial enzymes such...
متن کاملAtomic Resolution Crystal Structure of NAD+-Dependent Formate Dehydrogenase from Bacterium Moraxella sp. C-1
The crystal structure of the ternary complex of NAD+-dependent formate dehydrogenase from the methylotrophic bacterium Moraxella sp. C-1 with the cofactor (NAD+) and the inhibitor (azide ion) was established at 1.1 A resolution. The complex mimics the structure of the transition state of the enzymatic reaction. The structure was refined with anisotropic displacitalicents parameters for non-hydr...
متن کاملCloning , Nucleotide Sequencing , and Expression in Escherichia coti of the Gene for Formate Dehydrogenase of Paracoccus sp . 12 - A , a Formate - assimilating Bacterium
particular, with bacterial enzymes such as those of MOraxetla sp. <86.5%) and Rseudomonas sp. 101 (83.5%). The gene vvas highly expressed in Eycherichia coli cells using an expression plasmid with the pUC ori and tac promoter. The recombinant enzyme was somewhat inactiye in the stage of the cell-free extract, but its activity markedly increased with purification, in particular, with the step of...
متن کاملThe Role of Ala198 in the Stability and Coenzyme Specificity of Bacterial Formate Dehydrogenases
It has been shown by an X-ray structural analysis that the amino acid residues Ala198, which are located in the coenzyme-binding domain of NAD(+)-dependent formate dehydrogenases (EC 1.2.1.2., FDH) from bacteria Pseudomonas sp.101 and Moraxella sp. C-1 (PseFDH and MorFDH, respectively), have non-optimal values of the angles ψ and φ. These residues were replaced with Gly by site-directed mutagen...
متن کاملPurification and characterization of an acidic, thermophilic phytase from a newly isolated Geobacillus stearothermophilus strain DM12
Microbial phytases were applied mainly to animal and human foodstuffs in order to improvemineral bioavailability and food processing. In addition, phytases have potentialbiotechnological application in various other fields, such as environmental protection,aquaculture and agriculture. Bacillus sp. DM12, an isolate from a hot spring, produces phytase,which catalyzes the hydrolysis of phytic acid...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Journal of bacteriology
دوره 170 7 شماره
صفحات -
تاریخ انتشار 1988