Biochemical Characterization of Buffalo Liver Glucose-6-Phosphate Dehydrogenase Isoforms

2015 ◽  
Vol 34 (3) ◽  
pp. 193-204 ◽  
Author(s):  
Mahmoud A. Ibrahim ◽  
Abdel-Hady M. Ghazy ◽  
Ahmed M. H. Salem ◽  
Mohamed A. Ghazy ◽  
Mohammed M. Abdel-Monsef
Keyword(s):  
Biochemical Characterization ◽  
Glucose 6 Phosphate Dehydrogenase

scholarly journals Functional and Biochemical Characterization of Three Recombinant Human Glucose-6-Phosphate Dehydrogenase Mutants: Zacatecas, Vanua-Lava and Viangchan

2016 ◽  
Vol 17 (5) ◽  
pp. 787 ◽  
Author(s):  
Saúl Gómez-Manzo ◽  
Jaime Marcial-Quino ◽  
America Vanoye-Carlo ◽  
Hugo Serrano-Posada ◽  
Abigail González-Valdez ◽  
...  
Keyword(s):  
Biochemical Characterization ◽  
Glucose 6 Phosphate Dehydrogenase

scholarly journals Expression and biochemical characterization of human glucose-6- phosphate dehydrogenase in Escherichia coli: a system to analyze normal and mutant enzymes

Blood ◽  
1994 ◽  
Vol 83 (5) ◽  
pp. 1436-1441 ◽  
Author(s):  
TK Tang ◽  
CH Yeh ◽  
CS Huang ◽  
MJ Huang
Keyword(s):  
Escherichia Coli ◽  
Biochemical Characterization ◽  
Fold Increase ◽  
Binding Domain ◽  
Occurrence Rate ◽  
Substrate Analogues ◽  
Normal Enzyme ◽  
Glucose 6 Phosphate Dehydrogenase

We have developed a system to characterize normal and mutated glucose-6- phosphate dehydrogenase (G6PD) enzymes in vitro. Normal or mutant G6PD cDNA was subcloned into a pGEX-3X vector, which allowed production of a functional fusion protein in Escherichia coli. When we compared the recombinant normal enzyme with authentic human G6PD, indistinguishable Km values for glucose-6-phosphate (G6P) and NADP were obtained, and the utilization rates for two substrate analogues (2-deoxy G6P and deamino NADP) also showed no difference between the enzymes. This system was used to assay a biochemically uncharacterized variant, G6PD Taipei (493 A-->wG; 165 Asn-->Asp), plus two other known mutations (487 G-->A; 163 Gly-->Ser and 592 C-->T; 198 Arg-->Cys) that are located close to or within the putative G6P binding domain. Our results show that the G6PD activities of these three mutants were greatly reduced. No significant alteration in G6PD kinetics was observed for both 487 and 493 mutations. However, a drastic reduction in the Km for G6P (4-fold decrease) and tremendous increases in utilization rates of 2-deoxy G6P (32-fold increase) and deamino NADP (6-fold increase) were associated with the 592 mutation. This results suggests that arginine 198 in human G6PD, possibly located within the putative G6P binding domain, may play an important role in binding the substrate G6P. In addition, we and others have recently identified that at least nine different types of mutations are responsible for G6PD deficiency in Chinese. In this report, we also present the occurrence rate of each mutation present in the population of Taiwan.

scholarly journals Expression and biochemical characterization of human glucose-6- phosphate dehydrogenase in Escherichia coli: a system to analyze normal and mutant enzymes

Blood ◽  
1994 ◽  
Vol 83 (5) ◽  
pp. 1436-1441 ◽  
Author(s):  
TK Tang ◽  
CH Yeh ◽  
CS Huang ◽  
MJ Huang
Keyword(s):  
Escherichia Coli ◽  
Biochemical Characterization ◽  
Fold Increase ◽  
Binding Domain ◽  
Occurrence Rate ◽  
Substrate Analogues ◽  
Normal Enzyme ◽  
Glucose 6 Phosphate Dehydrogenase

Abstract We have developed a system to characterize normal and mutated glucose-6- phosphate dehydrogenase (G6PD) enzymes in vitro. Normal or mutant G6PD cDNA was subcloned into a pGEX-3X vector, which allowed production of a functional fusion protein in Escherichia coli. When we compared the recombinant normal enzyme with authentic human G6PD, indistinguishable Km values for glucose-6-phosphate (G6P) and NADP were obtained, and the utilization rates for two substrate analogues (2-deoxy G6P and deamino NADP) also showed no difference between the enzymes. This system was used to assay a biochemically uncharacterized variant, G6PD Taipei (493 A-->wG; 165 Asn-->Asp), plus two other known mutations (487 G-->A; 163 Gly-->Ser and 592 C-->T; 198 Arg-->Cys) that are located close to or within the putative G6P binding domain. Our results show that the G6PD activities of these three mutants were greatly reduced. No significant alteration in G6PD kinetics was observed for both 487 and 493 mutations. However, a drastic reduction in the Km for G6P (4-fold decrease) and tremendous increases in utilization rates of 2-deoxy G6P (32-fold increase) and deamino NADP (6-fold increase) were associated with the 592 mutation. This results suggests that arginine 198 in human G6PD, possibly located within the putative G6P binding domain, may play an important role in binding the substrate G6P. In addition, we and others have recently identified that at least nine different types of mutations are responsible for G6PD deficiency in Chinese. In this report, we also present the occurrence rate of each mutation present in the population of Taiwan.

Purification and characterization of glucose-6-phosphate dehydrogenase from Eisenia fetida and effects of some pesticides and metal ions

2020 ◽  
Vol 45 (4) ◽  
pp. 373-380
Author(s):  
Naciye Kayhan ◽  
Veysel Çomaklı ◽  
Sevki Adem ◽  
Caglar Güler
Keyword(s):  
Metal Ions ◽  
Eisenia Fetida ◽  
Biochemical Characterization ◽  
Inhibitory Effect ◽  
Biochemical Properties ◽  
Affinity Column ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Sepharose 4B

AbstractObjectivesEarthworms have a large impact on the soil ecosystem. They are quite sensitive to pollutants. Purification and biochemical characterization of glucose-6-phosphate dehydrogenases (G6PD) from the earthworm species Eisenia fetida were aimed. The determination of the toxicity potentials of some soil pollutants on G6PD activity was intended.MethodsG6PD was isolated using 2′,5′-ADP-Sepharose 4B affinity column. Enzyme purity and molecular mass were determined by SDS-PAGE. Its biochemical properties investigated. The effects of some soil pollutants on the enzyme were studied in vitro.ResultsEnzyme was purified with 28% yields and 232 fold. Optimum pH and buffer concentration, optimal and stable temperature was determined as pH: 8.5, 60 mM, 25 °C and 20 °C. Its molecular weight estimated as 36 kDa. The Ni2+, Hg2+, Pb2+, Cr2+, and Fe2+ ions with IC50 values in the range of 56 ± 06−120 ± 20 μM and the diniconazole, metalaxyl, methomyl, carboxyl, and oxamyl with IC50 values in the range of 7.6 ± 1.2−77 ± 12 μM exhibited an inhibitory effect on G6PD.ConclusionsG6PD was isolated and characterized from E. fetida. Its catalytic activity decreased with very low concentration by pesticides and metal ions. The results indicated that the inhibition of G6PD may be important in the toxicity mechanism of pollutants on this earthworm.

Cloning and biochemical characterization of three glucose‑6‑phosphate dehydrogenase mutants presents in the Mexican population

2018 ◽  
Vol 119 ◽  
pp. 926-936 ◽  
Author(s):  
Yadira Yazmín Cortés-Morales ◽  
America Vanoye-Carlo ◽  
Rosa Angélica Castillo-Rodríguez ◽  
Hugo Serrano-Posada ◽  
Abigail González-Valdez ◽  
...  
Keyword(s):  
Biochemical Characterization ◽  
Mexican Population ◽  
Glucose 6 Phosphate Dehydrogenase

Plant Pathology Diagnosed by X-Ray Microanalysis

1987 ◽  
Vol 45 ◽  
pp. 974-975
Author(s):  
J. H. Resau ◽  
N. Howell ◽  
S. H. Chang
Keyword(s):  
Electron Microscopy ◽  
Plant Pathology ◽  
Biochemical Characterization ◽  
Nutrient Deficiency ◽  
Green Leaf ◽  
Parasitic Infestation ◽  
X Ray

Spinach grown in Texas developed “yellow spotting” on the peripheral portions of the leaves. The exact cause of the discoloration could not be determined as there was no evidence of viral or parasitic infestation of the plants and biochemical characterization of the plants did not indicate any significant differences between the yellow and green leaf portions of the spinach. The present study was undertaken using electron microscopy (EM) to determine if a micro-nutrient deficiency was the cause for the discoloration.Green leaf spinach was collected from the field and sent by express mail to the EM laboratory. The yellow and equivalent green portions of the leaves were isolated and dried in a Denton evaporator at 10-5 Torr for 24 hrs. The leaf specimens were then examined using a JEOL 100 CX analytical microscope. TEM specimens were prepared according to the methods of Trump et al.

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