Complete nucleotide sequence of<i>Pseudomonas fluortscens</i>D-galactose dehydrogenase gene

Sperka, Sabine; Zehelein, E; Fiedler, S.; Fischer, Stephan; Sommer, Reinhold; Buckel, Peter

doi:10.1093/nar/17.13.5402

Cited by 11 publications

(9 citation statements)

References 2 publications

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“…The achieved data also demonstrated the applicability of the studied system for GalDH recovery. The subunit MW of recombinant GalDH was estimated to be about 34 kDa, which was similar to the reported value from P. fluorescens [5,6]. The K m for the purified recombinant GalDH was measured to be 0.32 mM.…”

Section: Sds-page and Kinetic Analysis Of Recombinant Galdhsupporting

confidence: 83%

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Recombinant d-galactose dehydrogenase partitioning in aqueous two-phase systems: effect of pH and concentration of PEG and ammonium sulfate

Kianmehr

Pooraskari

Mousavikoodehi

et al. 2014

Bioresour. Bioprocess.

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Background: D-Galactose dehydrogenase (GalDH; EC 1.1.1.48) belongs to the family of oxidoreductases that catalyzes the reaction of β-D-galactopyranose in the presence of NAD + to D-galacto-1,5-lactone and NADH. The enzyme has been used in diagnostic kits to neonatal screen for galactosemia diseases. This article reports the partitioning optimization of recombinant Pseudomonas fluorescens GalDH in aqueous two-phase systems (ATPS). Methods: Preliminary two-phase experiments exhibited that the polyethylene glycol (PEG) concentration, pH value, and concentration of salt had a significant influence on the partitioning efficiency of recombinant enzyme. According to these data, response surface methodology (RSM) with a central composite rotatable design (CCRD) was performed to condition optimization.

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Section: Sds-page and Kinetic Analysis Of Recombinant Galdhsupporting

confidence: 83%

“…The kinetic mechanism of Bi-Bi has been determined for this enzyme, with the NAD + binding first to the enzyme. The substrates of GalDH are D-galactose and NAD + , whereas its products are D-galactono-1,4-lactone, NADH, and H + [5]. GalDH has been identified in plants (e.g., green peas and Arabidopsis thaliana), algae (e.g.…”

Section: Introductionmentioning

confidence: 99%

Recombinant d-galactose dehydrogenase partitioning in aqueous two-phase systems: effect of pH and concentration of PEG and ammonium sulfate

Kianmehr

Pooraskari

Mousavikoodehi

et al. 2014

Bioresour. Bioprocess.

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“…High degrees of similarity to L-arabinose 1-dehydrogenase (over 100 bits of score) were found in bacterial (putative) oxidoreductases and dehydrogenases, including D-galactose 1-dehydrogenase from Pseudomonas fluorescens (22), confirming that L-arabinose 1-dehydrogenase is also a D-galactose 1-dehydrogenase. In the unrooted phylogenetic tree, these enzymes containing L-arabinose 1-dehydrogenase formed a single subfamily (Fig.…”

Section: Cloning Of the Gene Encoding L-arabinose 1-dehydrogenase Andmentioning

confidence: 82%

Cloning, Expression, and Characterization of Bacterial l-Arabinose 1-Dehydrogenase Involved in an Alternative Pathway of l-Arabinose Metabolism

Watanabe

Kodak

Makino

2006

Journal of Biological Chemistry

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“…The processing site of the GFOR presecretory protein between amino acid residues 52 and 53 is marked by an arrow. MocA and ORF334, putative rhizopine dehydrogenases of Rhizobium meliloti (24); DDP-DH, phtalic acid dehydrogenase from Pseudomonas putida (45); GalDH, galactose dehydrogenase from Pseudomonas fluorescens (46); StrI, putative protein in streptomycin biosynthesis from Streptomyces griseus (47); BvRed, biliverdin reductase from rat kidney (48); IDH, inositol dehydrogenase from B. subtilis (36); LmbZ, putative gene product of lmbZ of lincomycin biosynthesis from Streptomyces lincolnensis (49).…”

Section: Revised Gfo Sequence and Prediction Of The Secondarymentioning

confidence: 99%

The Substitution of a Single Amino Acid Residue (Ser-116 → Asp) Alters NADP-containing Glucose-Fructose Oxidoreductase ofZymomonas mobilis into a Glucose Dehydrogenase with Dual Coenzyme Specificity

Wiegert

Sahm

Sprenger

1997

Journal of Biological Chemistry

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Glucose-fructose oxidoreductase (GFOR, EC 1.1.1.99.-) from the Gram-negative bacterium Zymomonas mobilis contains the tightly bound cofactor NADP. Based on the revision of the gfo DNA sequence, the derived GFOR sequence was aligned with enzymes catalyzing reactions with similar substrates. A novel consensus motif (AGKHVXCEKP) for a class of dehydrogenases was detected. From secondary structure analysis the serine-116 residue of GFOR was predicted as part of a Rossmann-type dinucleotide binding fold. An engineered mutant protein (S116D) was purified and shown to have lost tight cofactor binding based on (a) altered tryptophan fluorescence; (b) lack of NADP liberation through perchloric acid treatment of the protein; and (c) lack of GFOR enzyme activity. The S116D mutant showed glucose dehydrogenase activity (3.6 ؎ 0.1 units/mg of protein) with both NADP and NAD as coenzymes (K m for NADP, 153 ؎ 9 M; for NAD, 375 ؎ 32 M). The single site mutation therefore altered GFOR, which in the wildtype situation contains NADP as nondissociable redox cofactor reacting in a ping-pong type mechanism, to a dehydrogenase with dissociable NAD(P) as cosubstrate and a sequential reaction type. After prolonged preincubation of the S116D mutant protein with excess NADP (but not NAD), GFOR activity could be restored to 70 units/mg, one-third of wild-type activity, whereas glucose dehydrogenase activity decreased sharply. A second site mutant (S116D/K121A/K123Q/I124K) showed no GFOR activity even after preincubation with NADP, but it retained glucose dehydrogenase activity (4.2 ؎ 0.2 units/mg of protein).

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Complete nucleotide sequence ofPseudomonas fluortscensD-galactose dehydrogenase gene

Cited by 11 publications

References 2 publications

Recombinant d-galactose dehydrogenase partitioning in aqueous two-phase systems: effect of pH and concentration of PEG and ammonium sulfate

Recombinant d-galactose dehydrogenase partitioning in aqueous two-phase systems: effect of pH and concentration of PEG and ammonium sulfate

Cloning, Expression, and Characterization of Bacterial l-Arabinose 1-Dehydrogenase Involved in an Alternative Pathway of l-Arabinose Metabolism

The Substitution of a Single Amino Acid Residue (Ser-116 → Asp) Alters NADP-containing Glucose-Fructose Oxidoreductase ofZymomonas mobilis into a Glucose Dehydrogenase with Dual Coenzyme Specificity

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