• BMB Reports
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• v.45 no.12
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• pp.707-712
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• 2012

Human glutamate dehydrogenase isozymes (hGDH1 and hGDH2) have been known to be inhibited by palmitoyl-CoA with a high affinity. In this study, we have performed the cassette mutagenesis at six different Cys residues (Cys59, Cys93, Cys119, Cys201, Cys274, and Cys323) to identify palmitoyl-CoA binding sites within hGDH2. Four cysteine residues at positions of C59, C93, C201, or C274 may be involved, at least in part, in the inhibition of hGDH2 by palmitoyl-CoA. There was a biphasic relationship, depending on the levels of palmitoyl-CoA, between the binding of palmitoyl-CoA and the loss of enzyme activity during the inactivation process. The inhibition of hGDH2 by palmitoyl-CoA was not affected by the allosteric inhibitor GTP. Multiple mutagenesis studies on the hGDH2 are in progress to identify the amino acid residues fully responsible for the inhibition by palmitoyl-CoA.

• Korean Journal of Plant Resources
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• v.10 no.4
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• pp.305-313
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• 1997

The effect of acid rain on plant isozyme response was studied with the treatment of simulated acid rain to Pinus densiflora, P. nigida and P. koraiensis for 9 months. The isozyme pattern of $\alpha$-Esterase($\alpha$-Est), Peroxidase(POD) and Glutamate dehydrogenase(GDH) were observed in the control (pH 5.6) and simulated acid rain (pH 3.5) treatment. No changes of isozyme pattern in $\alpha$-Est was observed in P. densiflora after 9 month treatment of simulated acid rain, but, two new isozymes were activated in P. nigida in the same treatment. In P. koraiensis, two new isozyme were activated but five isozymes were not activated. P. densiflora did not show any difference in POD and GDH after the treatment of simulated acid rain. P. nigida showed activation of eight and two isozymes in POD and GDH, respectively. P. koraiensis showed inactivation of 4 isozymes in POD but showed no changes In GDH.

• Journal of Microbiology and Biotechnology
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• v.24 no.11
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• pp.1516-1524
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• 2014

Flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH) can utilize a variety of external electron acceptors and also has stricter substrate specificity than any other glucose oxidoreductases, which makes it the ideal diagnostic enzyme in the field of glucose biosensors. A gene coding for a hypothetical protein, similar to glucose oxidase and derived from Aspergillus terreus NIH2624, was overexpressed in Pichia pastoris GS115 under the control of an AOX1 promoter with a level of 260,000 U/l in the culture supernatant after fed-batch cultivation for 84 h. After a three-step purification protocol that included isopropanol precipitation, affinity chromatography, and a second isopropanol precipitation, recombinant FAD-GDH was purified with a recovery of 65%. This is the first time that isopropanol precipitation has been used to concentrate a fermentation supernatant and exchange buffers after affinity chromatography purification. The purified FAD-GDH exhibited a broad and diffuse band between 83 and 150 kDa. The recombinant FAD-GDH was stable across a wide pH range (3.5 to 9.0) with maximum activity at pH 7.5 and $55^{\circ}C$. In addition, it displayed very high thermal stability, with a half-life of 82 min at $60^{\circ}C$. These characteristics indicate that FAD-GDH will be useful in the field of glucose biosensors.

• Journal of Life Science
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• v.18 no.6
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• pp.878-883
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• 2008

Aeromonas hydrophila DA33 was isolated from cultivated soils as a bacteria having high abilities to solubilize inorganic phosphate. Glucose dehydrogenase gene (gdh) was cloned from Escherichia coli. The recombinant plasmid, pGHS containing glucose dehydrogenase gene was introduced into A. hydrophila DA33 in order to improve the activity of phosphate-solubilizing. The transformant harboring the gdh gene, A. hydrophila pGHS/DA33 increased enzyme activity. The strain also increased the gluconic acid generation that was effective for phosphate solubilization. It was possible that the strain containing pGHS produced higher solubilized phosphate with tri-calcium phosphate as the unique (P) source, in comparison with that of wild type without plasmid. These results suggest that the strain, A. hydrophila pGHS/DA33 is expected as effective biofertilizer for phosphate solubilization.

• The Korean Journal of Zoology
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• v.12 no.2
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• pp.50-56
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• 1969

The present investigation involves a comparative study of enzymatic activities in various animals. The levels of the liver protein of rat (22.0$\pm$0.01mg/ml and pigeon (22.0$\pm$ 0.16) are twice as high as that of crucian carp (13.0$\pm$ 0.09)(p < 0.01). Generally, the specific acitivity (3.77 $\pm$0.18 unit/mg) of rat glutamic pyruvic transaminase (GPT) is highest, pigeon intermediate (1.93 $\pm$0.01), and crucian carp lowest (0.71$\pm$0.07). On the other hand, the specific activity (8.23$\pm$0.09 unit/mg)of rat glutamic oxaloacetic transaminase (GOT) is highest, pigeon intermediate (3.95$\pm$0.09), and crucian carp lowest (0.92$\pm$0.01) (p < 0.01). Ratios of GOT activity to GPT activity appear no remarkable difference from the levels of various animals. Specific activity of glutamic dehydrogenase(GDH) in pigeon tissue exceeds those of rat and crucian carp. In liver, rat GOT specific activity is greater than crucian carp and pigeon. On the other hand, pigeon GDH specific activity is greater than those of rat and crucian carp. This wouls seem to be in accord with protein metabolic intensity. The patterns for GDH isozyme were remarkably appeared in various animals. Glutamic dehydrogenase isozymes gave different electrophoretic mobilities in various animals. It is interesting that crucian carp, pigeon, and rat would show this difference, which may be indicative of an evolutional pattern. The fact that livers in various animals show quite different enzyme activities would suggest the existence of such a general phylogenetic relationship.

• Journal of Nutrition and Health
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• v.4 no.1
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• pp.21-28
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• 1971

1. The effects of chloroform to the tissue lactic dehydrogenase (LDH) activities and its isozymes and to the tissue glutamic dehydrogenase (GDH) activities and its isoaymes are studied using the experimental albino male adult rats in this paper. The tissues studies are liver, kidney, heart, and brain. Besides the control group, two experimental groups are studied providing succeedingly 4 days interpariental administrations of chloroform, 0.0025ml and 0.025ml per day respectively. The changes of body weights, weights of organs, activities of GDH and LDH and their isozymes of each tissues, are analysed. 2. The body weights of rats are decreased due to the chloroform administration. 3. There are no significant differences of weights of organs due to the chloroform administration. 4. The significant decreases of tissue GDH activities and the significant changes in percent distribution of the GDH isozymes are found due to the chloroform administration. This weight be interpretated that chloroform effects to the protein and amino acid metabolism of rats. 5. Due to the chloroform administration, the significant changes in tissue LDH activities and in percent distribution of tissue LDH isozymes indicating the decreases of $LDH_1$ which is the aerobic heart type and the increase of $LDH_5$ which is the anaerobic muscle type, are observed. This could be estimated that chloroform effects to the carbohydrate metabolism, particularly to the anaerobic glycolysis of rats.

• Molecules and Cells
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• v.19 no.1
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• pp.97-103
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• 2005

The role of residue C323 in catalysis by human glutamate dehydrogenase isozymes (hGDH1 and hGDH2) was examined by substituting Arg, Gly, Leu, Met, or Tyr at C323 by cassette mutagenesis using synthetic human GDH isozyme genes. As a result, the $K_m$ of the enzyme for NADH and ${\alpha}-ketoglutarate$ increased up to 1.6-fold and 1.1-fold, respectively. It seems likely that C323 is not responsible for substrate-binding or coenzyme-binding. The efficiency ($k_{cat}/K_m$) of the mutant enzymes was only 11-14% of that of the wild-type isozymes, mainly due to a decrease in $k_{cat}$ values. There was a linear relationship between incorporation of [$^{14}C$]p-chloromercuribenzoic acid and loss of enzyme activity that extrapolated to a stoichiometry of one mol of [$^{14}C$] incorporated per mol of monomer for wild type hGDHs. No incorporation of [$^{14}C$]p-chloromercuribenzoic acid was observed with the C323 mutants. ADP and GTP had no effect on the binding of p-chloromercuribenzoic acid, suggesting that C323 is not directly involved in allosteric regulation. There were no differences between the two hGDH isozymes in sensitivities to mutagenesis at C323. Our results suggest that C323 plays an important role in catalysis by human GDH isozymes.

• The Korean Journal of Ecology
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• v.14 no.1
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• pp.75-85
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• 1991

Changes in the activities of isocitrate dehydrogenase (IDH) and glutamate dehydrogenase (GDH) and changes in free amino acids in the cytoplasm of Saccharomyces cerevisiae have been studied under heat shock condition. Heat shock conditions led to a significant decrease of NAD-IDH and NAD-GDH, It was shown appeared that the meaningful patterns of increase of NADP-IDH and NADP-GDH. It suggested that heat shock in yeast leads to a splitting of the TCA cycle and that glutamate synthesis takes place through the coupling of the NADP-linked isocirate and glutamate dehydrogenase. It was shown that about 14% of total free amino acids of yeast cells was decreased by heat shock. Especially heat shock condition resulted in the marked decreases of serine family amino acids such as serine, glycine and cysteine, and in the considerable increases of the rates of methionine, alanine, glutamin.

• Journal of the Korean Electrochemical Society
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• v.17 no.1
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• pp.30-36
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• 2014

Redox complexes to transport electrons from enzyme to electrodes are very important part in glucose sensor. Pentacyanoferrate-bound aniline ($Fe(CN)_5$-aminopyridine), was prepared as a potential redox mediator in a glucose dehydrogenase (GDH)-glucose sensor. The synthesized pyridyl-$NH_2$ to pentacyanoferrate was characterized by the electrochemical and spectroscopic methods. A amperometric enzyme-linked electrode was developed based on GDH, which catalyses the oxidation of glucose. Glucose was detected using GDH that was co-immobilized with an $Fe(CN)_5$-aminopyridine and gold nano-particles (AuNPs) on ITO electrodes. The $Fe(CN)_5$-aminopyridine and AuNPs immobilized onto ITO electrodes provided about a two times higher electrochemical response compared to that of a bare ITO electrode. As glucose was catalyzed by wired GDH, the electrical signal was monitored at 0.4 V versus Ag/AgCl by cyclic voltammetry. The anode currents was linearly increased in proportion to the glucose concentration over the 0~10 mM range.

• Journal of Korean Society of Forest Science
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• v.66 no.1
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• pp.109-117
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• 1984

Allozyme study for open pollinated seeds of forty nine pitch pine families in an $F_1$-hybrid seed orchard demonstrated that allozyme variants in aspartate aminotransferase(AAT), glutamate dehydrogenase(GDH) and leucine aminopeptidase(LAP) systems are encoded by at Least eight loci; five for AAT, one for GDH and two fur LAP. Allozyme variations showed polymorphisms at seven of the eight loci, except GDH. Average number of alleles examined over six loci were 2.33 and 2.67 for maternal and progeny groups, respectively. Average heterozygosity and genetic diversity computed over six loci were, respectively, 0.235 and 5.409 for maternal tree group, 0.238 and 5.569 for progeny group. The proportion of $F_1$-hybrid seeds estimated by allozyme analysis was 0.77%. The estimated proportion of $F_1$ hybrid seeds by allozyme study is in good agreement with the value 0.73% estimated by morphological study for the proportion of pitch ${\times}$ Loblolly $F_1$ hybrid seedlings at a nursery. Indications for Wahlund effect, high levels of self-fertilization and for non-random matings in the $F_1$ hybrid seed orchard call for cautions in estimating allele frequency changes and mating probabilities for the parental and progeny groups.