A comparative study of the thermal stability of formate dehydrogenases from microorganisms and plants

Садыхов, Э. Г.; Serov, A. E.; Voinova, N. S.; Uglanova, S. V.; Petrov, A. S.; Алексеева, А. А.; Kleimenov, S. Yu.; Popov, Vladimir O.; Тишков, В. И.

doi:10.1134/s0003683806030021

Cited by 31 publications

(16 citation statements)

References 10 publications

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“…They suggest that both NADP + and formate stabilize the enzyme slightly more than NAD + . All these values fall in the range of many other FDHs reported so far (50-68°C) while compare unfavorably with those reported for Pseudomonas, Moraxella, and C. boidinii (varying from 68 to 58°C), all measured at pH 7 (Sadykhov et al 2006).…”

Section: Rt)supporting

confidence: 63%

“…Considering the chance of using the new FDH in biocatalysis, we proceeded to examining its stability to temperature and cosolvents, as prerequisite information to applicability (Sadykhov et al 2006;Tishkov and Popov 2006). The activity as a function of temperature was studied to determine the kinetic constants for the Arrhenius equation (K cat = A · e (−Ea/…”

Section: Evaluation Of Enzyme Stabilitymentioning

confidence: 99%

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Structural basis for double cofactor specificity in a new formate dehydrogenase from the acidobacterium Granulicella mallensis MP5ACTX8

Fogal

Beneventi

Cendron

et al. 2015

Appl Microbiol Biotechnol

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Formate dehydrogenases (FDHs) are considered particularly useful enzymes in biocatalysis when the regeneration of the cofactor NAD(P)H is required, that is, in chiral synthesis with dehydrogenases. Their utilization is however limited to the recycling of NAD(+), since all (apart one) of the FDHs characterized so far are strictly specific for this cofactor, and this is a major drawback for their otherwise wide applicability. Despite the many attempts performed to modify cofactor specificity by protein engineering different NAD(+)-dependent FDHs, in the general practice, glucose or phosphite dehydrogenases are chosen for the recycling of NADP(+). We report on the functional and structural characterization of a new FDH, GraFDH, identified by mining the genome of the extremophile prokaryote Granulicella mallensis MP5ACTX8. The new enzyme displays a valuable stability in the presence of many organic cosolvents as well as double cofactor specificity, with NADP(+) preferred over NAD(+) at acidic pH values, at which it also shows the highest stability. The quite low affinities for both cofactors as well as for the substrate formate indicate, however, that the native enzyme requires optimization to be applied as biocatalytic tool. We also determined the crystal structure of GraFDH both as apoprotein and as holoprotein, either in complex with NAD(+) or NADP(+). Noticeably, the latter represents the first structure of an FDH enzyme in complex with NADP(+). This fine picture of the structural determinants involved in cofactor selectivity will possibly boost protein engineering of the new enzyme or other homolog FDHs in view of their biocatalytic exploitation for NADP(+) recycling.

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Section: Rt)supporting

confidence: 63%

Section: Evaluation Of Enzyme Stabilitymentioning

confidence: 99%

Structural basis for double cofactor specificity in a new formate dehydrogenase from the acidobacterium Granulicella mallensis MP5ACTX8

Fogal

Beneventi

Cendron

et al. 2015

Appl Microbiol Biotechnol

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“…For example, the rate of thermal inactivation of the Glu170Asp mutant of PseFDH is 20% lower than that of full-length PseFDH (Tishkov, 2009). However, a comparison of the DSC data shows that the melting point of MorFDH (336.4 K) is 4.5 K lower than that of PseFDH (Sadykhov et al, 2006). However, as can be seen from the DSC curves, the thermal denaturation of MorFDH and PseFDH is a one-step highly cooperative process that does not implicate the dissociation of the enzyme molecules into subunits in intermediate steps.…”

Section: Tablementioning

confidence: 92%

Structures of the apo and holo forms of formate dehydrogenase from the bacteriumMoraxellasp. C-1: towards understanding the mechanism of the closure of the interdomain cleft

Shabalin

Filippova

Polyakov

et al. 2009

Acta Crystallogr D Biol Cryst

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NAD(+)-dependent formate dehydrogenase (FDH) catalyzes the oxidation of formate ion to carbon dioxide coupled with the reduction of NAD(+) to NADH. The crystal structures of the apo and holo forms of FDH from the methylotrophic bacterium Moraxella sp. C-1 (MorFDH) are reported at 1.96 and 1.95 A resolution, respectively. MorFDH is similar to the previously studied FDH from the bacterium Pseudomonas sp. 101 in overall structure, cofactor-binding mode and active-site architecture, but differs in that the eight-residue-longer C-terminal fragment is visible in the electron-density maps of MorFDH. MorFDH also differs in the organization of the dimer interface. The holo MorFDH structure supports the earlier hypothesis that the catalytic residue His332 can form a hydrogen bond to both the substrate and the transition state. Apo MorFDH has a closed conformation of the interdomain cleft, which is unique for an apo form of an NAD(+)-dependent dehydrogenase. A comparison of the structures of bacterial FDH in open and closed conformations allows the differentiation of the conformational changes associated with cofactor binding and domain motion and provides insights into the mechanism of the closure of the interdomain cleft in FDH. The C-terminal residues 374-399 and the substrate (formate ion) or inhibitor (azide ion) binding are shown to play an essential role in the transition from the open to the closed conformation.

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“…18 As a result, a mutant form of PseFDH was obtained, which exceeded the initial enzyme 100 fold in stability. 26 II. Increase in chemical stability.…”

Section: Methodsmentioning

confidence: 99%

Creation of biocatalysts with prescribed properties

2008

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Oxidoreductases are widely used in different branches of industry (particularly, in pharma ceutical) and analytical biotechnology. To optimize the use and to reduce production cost of a target product, the properties of a biocatalyst should be modified to meet requirements of a specific process or method of analysis. Problem of creation of biocatalysts with prescribed properties includes complex set of R&D such as an engineering of enzyme properties, con struction of recombinant strains overproducing the target protein, development of a large scale downstream process and preparation of a working form of a biocatalysts. In the present work, the basic features and practical examples of such an over all approach are considered for formate dehydrogenase and D amino acid oxidase.

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A comparative study of the thermal stability of formate dehydrogenases from microorganisms and plants

Cited by 31 publications

References 10 publications

Structural basis for double cofactor specificity in a new formate dehydrogenase from the acidobacterium Granulicella mallensis MP5ACTX8

Structural basis for double cofactor specificity in a new formate dehydrogenase from the acidobacterium Granulicella mallensis MP5ACTX8

Structures of the apo and holo forms of formate dehydrogenase from the bacteriumMoraxellasp. C-1: towards understanding the mechanism of the closure of the interdomain cleft

Creation of biocatalysts with prescribed properties

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