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Cloning and characterization of Lotus japonicus formate dehydrogenase: A possible correlation with hypoxia

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dc.contributor.author Andreadeli, A en
dc.contributor.author Flemetakis, E en
dc.contributor.author Axarli, I en
dc.contributor.author Dimou, M en
dc.contributor.author Udvardi, MK en
dc.contributor.author Katinakis, P en
dc.contributor.author Labrou, NE en
dc.date.accessioned 2014-06-06T06:49:38Z
dc.date.available 2014-06-06T06:49:38Z
dc.date.issued 2009 en
dc.identifier.issn 15709639 en
dc.identifier.uri http://dx.doi.org/10.1016/j.bbapap.2009.02.009 en
dc.identifier.uri http://62.217.125.90/xmlui/handle/123456789/4698
dc.subject Formate dehydrogenase en
dc.subject Hypoxia en
dc.subject L. japonicus en
dc.subject Nodule en
dc.subject Rhizobium en
dc.subject.other formate dehydrogenase en
dc.subject.other affinity chromatography en
dc.subject.other article en
dc.subject.other enzyme kinetics en
dc.subject.other enzyme purification en
dc.subject.other enzyme specificity en
dc.subject.other Escherichia coli en
dc.subject.other eukaryote en
dc.subject.other gene en
dc.subject.other gene expression regulation en
dc.subject.other hypoxia en
dc.subject.other legume en
dc.subject.other LjFDH1 gene en
dc.subject.other Lotus en
dc.subject.other lotus japonicus en
dc.subject.other molecular cloning en
dc.subject.other molecular model en
dc.subject.other nitrogen fixation en
dc.subject.other nonhuman en
dc.subject.other nucleotide sequence en
dc.subject.other priority journal en
dc.subject.other prokaryote en
dc.subject.other Pseudomonas en
dc.subject.other real time polymerase chain reaction en
dc.subject.other reverse transcription polymerase chain reaction en
dc.subject.other RNA RNA hybridization en
dc.subject.other Amino Acid Sequence en
dc.subject.other Anoxia en
dc.subject.other Base Sequence en
dc.subject.other Cloning, Molecular en
dc.subject.other DNA Primers en
dc.subject.other Formate Dehydrogenases en
dc.subject.other In Situ Hybridization en
dc.subject.other Lotus en
dc.subject.other Molecular Sequence Data en
dc.subject.other Polymerase Chain Reaction en
dc.subject.other Sequence Homology, Amino Acid en
dc.subject.other Eukaryota en
dc.subject.other Lotus corniculatus var. japonicus en
dc.subject.other Prokaryota en
dc.subject.other Pseudomonas sp. 101 en
dc.subject.other Rhizobium en
dc.title Cloning and characterization of Lotus japonicus formate dehydrogenase: A possible correlation with hypoxia en
heal.type journalArticle en
heal.identifier.primary 10.1016/j.bbapap.2009.02.009 en
heal.publicationDate 2009 en
heal.abstract Formate dehydrogenases (FDHs, EC 1.2.1.2) comprise a group of enzymes found in both prokaryotes and eukaryotes that catalyse the oxidation of formate to CO2. FDH1 from the model legume Lotus japonicus (LjFDH1) was cloned and expressed in E. coli BL21(DE3) as soluble active protein. The enzyme was purified using affinity chromatography on Cibacron blue 3GA-Sepharose. The enzymatic properties of the recombinant enzyme were investigated and the kinetic parameters (Km, kcat) for a number of substrates were determined. Molecular modelling studies were also employed to create a model of LjFDH1, based on the known structure of the Pseudomonas sp. 101 enzyme. The molecular model was used to help interpret biochemical data concerning substrate specificity and catalytic mechanism of the enzyme. The temporal expression pattern of LjFDH1 gene was studied by real-time RT-PCR in various plant organs and during the development of nitrogen-fixing nodules. Furthermore, the spatial transcript accumulation during nodule development and in young seedpods was determined by in situ RNA-RNA hybridization. These results considered together indicate a possible role of formate oxidation by LjFDH1 in plant tissues characterized by relative hypoxia. © 2009 Elsevier B.V. en
heal.journalName Biochimica et Biophysica Acta - Proteins and Proteomics en
dc.identifier.issue 6 en
dc.identifier.volume 1794 en
dc.identifier.doi 10.1016/j.bbapap.2009.02.009 en
dc.identifier.spage 976 en
dc.identifier.epage 984 en


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