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Gul-Karaguler, Nevin; Sessions, Richard B.; Clarke, Anthony R.; Holbrook, J. John

doi:10.1023/a:1005610414179

Cited by 51 publications

(18 citation statements)

References 14 publications

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“…The (k cat /K m ) NADP + value of this mutant FDH was only threefold less than the (k cat /K m ) NAD + value for the wild type enzyme [64]. The Asp195Ser substitution in FDH from C. methylica increased 10,000 fold the affinity of the enzyme for NADP + [63]; however, the enzyme was still much more specific to NAD + than to NADP + ( Table 2. Kinetic properties of mutant formate dehydrogenases and recombinant wild type enzymes from the yeast Saccharomyces cerevisiae (SceFDH), Candida methylica (CmeFDH), and the bacterium Pseudomonas sp.…”

Section: Specific Features Of the Catalytic Mechanism Of Formate Dehymentioning

confidence: 92%

“…11 2004 residue in the conservative triad (Gly/Ala)XGlyXXGly and the residue Asp221 (numeration according to the sequence of FDH from Pseudomonas sp. 101), which plays an important role in providing the specificity to NAD + [63,64]. Two other breaks are located between two pairs of catalytically essential residues Ile122 Asn146 and Gln313 His332 (Fig.…”

Section: Comparative Analysis Of Amino Acid Sequencesmentioning

confidence: 99%

“…The ratio between the k cat /K m values of this enzyme for NAD + and NADP + is ≥ 3·10 9 [64] (Table 2). FDH from the yeast C. methylica is also highly specific to NAD + ((k cat /K m ) NAD + /(k cat /K m ) NADP + > 250,000) [63]. The coenzyme selectivity of bacterial FDHs is much lower.…”

Section: Specific Features Of the Catalytic Mechanism Of Formate Dehymentioning

confidence: 99%

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Catalytic mechanism and application of formate dehydrogenase

Тишков

Popov

2004

Biochemistry (Moscow)

159

129

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NAD+-dependent formate dehydrogenase (FDH) is an abundant enzyme that plays an important role in energy supply of methylotrophic microorganisms and in response to stress in plants. FDH belongs to the superfamily of D-specific 2-hydroxy acid dehydrogenases. FDH is widely accepted as a model enzyme to study the mechanism of hydride ion transfer in the active center of dehydrogenases because the reaction catalyzed by the enzyme is devoid of proton transfer steps and implies a substrate with relatively simple structure. FDH is also widely used in enzymatic syntheses of optically active compounds as a versatile biocatalyst for NAD(P)H regeneration consumed in the main reaction. This review covers the late developments in cloning genes of FDH from various sources, studies of its catalytic mechanism and physiological role, and its application for new chiral syntheses.

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Section: Specific Features Of the Catalytic Mechanism Of Formate Dehymentioning

confidence: 92%

Section: Comparative Analysis Of Amino Acid Sequencesmentioning

confidence: 99%

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Catalytic mechanism and application of formate dehydrogenase

Тишков

Popov

2004

Biochemistry (Moscow)

159

129

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“…The improvements in k cat render PTDH very competitive in terms of turnover rate with the three most commonly used FDH enzymes from Candida boidinii, C. methylica, and an NADP specific mutant from Pseudomonas sp.101, which have kcat values of 240 min -1 [42], 84 min -1 [43], and 300 min -1 [29], respectively, with NAD as the cofactor. The final PTDH mutant has a higher kcat value (340 min -1 ) than the above-mentioned FDH enzymes.…”

Section: Kinetics Of Purified Mutant Ptdhsmentioning

confidence: 99%

Optimizing a Biocatalyst for Improved NAD(P)H Regeneration: Directed Evolution of Phosphite Dehydrogenase

Woodyer¹,

Donk²,

Zhao³

2006

CCHTS

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Cofactor regeneration is an important solution to the problem of implementing complex cofactor requiring enzymatic reactions at the industrial scale. NAD(P)H-dependent oxidoreductases are highly valuable biocatalysts, but the high cost of the nicotinamide cofactors necessitates in situ cofactor regeneration for preparative applications. Here we report the use of directed evolution to enhance the industrially important properties of phosphite dehydrogenase for NAD(P)H regeneration. A two-tiered sorting method of selection and screening was used in conjunction with random and rational mutagenesis. Following six rounds of directed evolution, soluble expression in E. coli was increased more than 3-fold, while the turnover rate was increased about 2-fold, effectively lowering the cost of the enzyme by >6-fold. Large-scale production of the final mutant enzyme by fermentation resulted in approximately 6-times higher yield (Units/Liter) than the WT enzyme. The enhancements of PTDH were independent of expression vector and E. coli strain utilized. The advantage of the final mutant over the WT enzyme was demonstrated using the industrially relevant bioconversion of trimethylpyruvate to L-tert-leucine. The mutations discovered are discussed in the context of a three dimensional structural model and the resulting changes in kinetics and soluble expression. The engineered phosphite dehydrogenase has great potential for NAD(P)H regeneration in industrial biocatalysis.

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“…Biological CO2 fixation systems have also received much attention. For example, CO2 or HCO3 − can be reduced to formic acid with formate dehydrogenase (FDH) and NADH 9) . Therefore, a CO2 fixation system that combines the photoreduction of NAD + by the photosensitization of zinc porphyrin and ferredoxin-NADP + reductase, and HCO3 − reduction with FDH as shown in Scheme 1 a) can be established.…”

Section: Introductionmentioning

confidence: 99%