A rapid method for the purification of glucose-6-phosphate dehydrogenase from bovine lens

Ulusu, Nuriye Nuray; Kus, M. Sezai; Açan, N. Leyla; Tezcan, E. Ferhan

doi:10.1016/s1357-2725(99)00019-9

Cited by 37 publications

(50 citation statements)

References 23 publications

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“…factors such as; dilution of the enzyme sample, ionic factors (salt) or physical effects. The molecular weight of the G6PD from sheep brain cortex was in accordance with our previous studies [6][7][8].…”

Section: Mechanism Kinetic Values Loss Valuesupporting

confidence: 89%

“…We have previously described a purification protocol for G6PD from various tissues; bovine lens, sheep and lamb kidney cortex [6][7][8]. In this study, we have determined the kinetic mechanism and few properties of the G6PD which was purified from sheep brain cortex [5].…”

Section: Resultsmentioning

confidence: 99%

“…G6PD was purified as previously described (5)(6)(7)(8). The method included 2', 5'-ADP-Sepharose 4B affinity chromatography and DEAE Sepharose Fast Flow ion exchange chromatographies after ultracentrifugation.…”

Section: Enzymementioning

confidence: 99%

“…G6PD catalyzes the transformation of G6P to 6-phosphogluconolactone and reduction of co-enzyme nicotinamide adenine dinucleotide phosphate to NADPH by different kinetic mechanisms in different organisms. It has been shown that G6PD catalyzes the reaction by ordered Bi Bi mechanism in Schizosaccharomyces pombe [24] mouse liver [25] lamb kidney cortex [26] Corynebacterium glutamicum [27] Acetobacter hansenii (formerly known as Acetobacter xylinum) [28] bovine lens [6]. Random Bi Bi mechanism is investigated in Plasmodium falciparum [29], Taenia crassiceps (Eucestoda: Cyclophyllidea) [30], Aspergillus aculeatus, a filamentous fungus [31], Aspergillus niger and Aspergillus nidulans [32].…”

Section: Kinetic Mechanism Productmentioning

confidence: 99%

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Kinetic mechanism and some properties of glucose-6-phosphate dehydrogenase from sheep brain cortex

Ulusu

Sengezer

2012

Turk J Bioch

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Section: Mechanism Kinetic Values Loss Valuesupporting

confidence: 89%

Section: Resultsmentioning

confidence: 99%

Section: Enzymementioning

confidence: 99%

Section: Kinetic Mechanism Productmentioning

confidence: 99%

See 2 more Smart Citations

Kinetic mechanism and some properties of glucose-6-phosphate dehydrogenase from sheep brain cortex

Ulusu

Sengezer

2012

Turk J Bioch

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“…The minimum structure necessary for catalysis is the dimeric form (Levy, 1979). But Ulusu et al (1999) showed that bovine lens G6PD can form a monomer in active state. Similar results were also found for alveolar nodule and adenocarcinoma G6PD in mammary glands of BALB/c mice (Ulusu et al, 1999).…”

Section: Resultsmentioning

confidence: 99%

Purification and characterization of glucose 6-phosphate dehydrogenase from rainbow trout (oncorhynchus mykiss) erythrocytes

Çiftçi¹,

Çiltaş²,

Erdoğan³

2004

Vet. Med. - Czech

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Glucose 6-phosphate dehydrogenase (D-glucose 6-phosphate: NADP + oxidoreductase, EC 1.1.1.49; G6PD) from rainbow trout (Oncorhynchus mykiss) erythrocytes was purified, using a simple and rapid method, and some characteristics of the enzyme were investigated. The purification procedure consisted of three steps: haemolysate preparation, ammonium sulphate precipitation, and 2',5'-ADP Sepharose 4B affinity gel chromatography, which took one working day. Thanks to the three consecutive procedures, the enzyme, having the specific activity of 14.51 EU/mg proteins, was purified with a yield of 70.40% and 1 271.19-fold. In order to control the purification of the enzyme SDS polyacrylamide gel electrophoresis was carried out. SDS polyacrylamide gel electrophoresis showed a single band for the enzyme. Optimal pH, stable pH, optimal temperature, molecular weight, and K M and V max values for NADP + and glucose 6-phosphate (G6-P) were also determined for the enzyme. In addition, the effect of NADPH on the enzyme was investigated and K i value and the type of inhibition were determined by means of Lineweaver-Burk graph obtained for NADPH.

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Purification and characterization of glucose 6-phosphate dehydrogenase from Lake Van fish (Chalcalburnus tarichii pallas, 1811) liver

Türkoğlu

Altun

Çiftçi

2006

J. Physiol. Biochem.

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Glucose 6-phosphate dehydrogenase (D-glucose 6-phosphate: NADP+ oxidoreductase, EC 1.1.1.49; G6PD) was purified from Lake Van fish (Chalcalburnus tarichii pallas, 1811) liver, using a simple and rapid method, and some characteristics of the enzyme were investigated. The purification procedure was composed of two steps: homogenate preparation and 2', 5'-ADP Sepharose 4B affinity gel chromatography, which took 7-8 hours. Thanks to the two consecutive procedures, the enzyme, having specific activity of 38 EU/mg protein, was purified with a yield of 44.39% and 1310 fold. In order to control the enzyme purification SDS polyacrylamide gel electrophoresis (SDS-PAGE) was done. SDS polyacrylamide gel electrophoresis showed a single band for enzyme. Optimal pH, stable pH, optimal temperature, Km and, Vmax values for NADP+ and glucose 6-phosphate (G6P) were also determined for the enzyme. In addition, molecular weight and subunit molecular weights were found by sodium dodecyl sulfate polyacrilamide gel electrophoresis (SDS-PAGE) and gel filtration chromatography respectively.

show abstract

A rapid method for the purification of glucose-6-phosphate dehydrogenase from bovine lens

Cited by 37 publications

References 23 publications

Kinetic mechanism and some properties of glucose-6-phosphate dehydrogenase from sheep brain cortex

Kinetic mechanism and some properties of glucose-6-phosphate dehydrogenase from sheep brain cortex

Purification and characterization of glucose 6-phosphate dehydrogenase from rainbow trout (oncorhynchus mykiss) erythrocytes

Purification and characterization of glucose 6-phosphate dehydrogenase from Lake Van fish (Chalcalburnus tarichii pallas, 1811) liver

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