• Korean Journal of Microbiology
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• v.26 no.3
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• pp.197-206
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• 1988

An outer membrane-associated 2-furaldehyde dehydrogenase, catalyzing the oxidation of 2-furaldehyde to 2-furoic acid from Klebsiella pneumoniae was purified to homogeneity and characterized. The enzyme showed its highly specific dependency on $\beta$-$NAD^{+}$. Enzyme activity was monitored during purification by using substrate 2-furaldehyde and coenzyme $\beta$-$NAD^{+}$ by means of high performance liquid chromatography. The outer membrane was successfully collected by the methods of Percoll density gradient ultracentrifugation and ultracentrifugation after preferential solubilization of the membrane with $Mg^{2+}$ and Triton X-100. The enzyme was purified by the series of procedures including extraction of outer membrane protein with EDTA and lysozume, and fractionation by column chromatography on QAE-Sephades Q-50, and subsequently Sephadex G-100. The enzume showed its optimal activity at $85^{\circ}C$, pH 9.5, and in the presence of 1.5% (vol/vol) Triton X-100. The enzyme exhibited a native molecular size of 88,000 by nondenaturing polyacrylamide gel electrophoresis and had an apparent Km of 4.72mM for 2-furaldehyde.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.27 no.1
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• pp.83-88
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• 1994

For the purpose of genetic stock indentification of three species of salmonid fishs and their hybrid, lactate dehydrogenase(LDH), malate dehydrogenase(MDH), isocitrate dehydrogenase(IDH), a-gylycerophosphate dehydrogenase(a-GPDH), malic enzyme(ME), 6-phospho-gluconate dehydrogenase(6-PGD), phosphoglucose isomerase(PGI) and phospho-glucomutase(PGM) from skeletal muscle, liver, heart and gill tissues in all three species were analyzed. Chum and masu salmon showed no polymorphic patterns in all isozyme loci, however rainbow trout were found to have polymorphic patterns at MDH-B, LDH and IDH loci. Especially, significant differences were found at MDH-B loci between the three species and the IDH patterns of rainbow trout were also different from the other two species. These loci therefore can be utilized as efficient genetic markers for the identification of hybrids and improve the efficiency of fish breeding. There was no difference except PGI between diploid and triploid isozyme patterns but PGI showed some potential as a marker for triploid in masu salmon.

• BMB Reports
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• v.29 no.6
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• pp.493-499
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• 1996

The biochemical properties of purified D-hydruxyisovalerate dehydrogenase from Fusarium sambucinum was elucidated. D-Hydroxyisovalerate dehydrogenase produced solely D-hydroxyisovalerate from 2-ketoisovalerate. The isoelectric point of the purified enzyme was 7.0. The enzyme was highly specific with 2-ketoisovalerate ($K_{m}=0.188$ mM, $V_{max}=8.814$ mmol/min mg) and 2-keto-3-methyl-n-valerate ($K_{m}=0.4$ mM, $V_{max}=1.851$ mmol/min mg) for the reductive reaction. This was also seen by comparing D-hydroxyisovalerate ($K_{m}=1.667$ mM, $V_{max}=0.407$ mmol/min mg) and D-hydroxy-3-methyl-n-valerate ($K_{m}=6.7$ mM, $V_{max}=0.648$ mmol/min mg) for the oxidative reaction. Thiol blocking reagents, such as iodoacetamide, N-ethylmaleimide and p-chloromecuribenzoate inhibited about 80% of enzyme activity at 0.02 mM, 50 mM and 50 mM, respectively. The enzyme activity was also inhibited by the addition of 0.1 mM of various metal ions, such as $Fe^{2+}$ (67%), $Cu^{2+}$ (88%), $Zn^{2+}$ t (76%) and $Mg^{2+}$ (9%). The enzyme was stable over three months in 50 mM potassium phosphate buffer (pH 5~7) at $-80^{\circ}C$. However the purified enzyme lost 30% of its activity in the same buffer after 24 h at $4^{\circ}C$. The studies about thermal inactivation of D-hydroxyisovalerate dehydrogenase exhibit 209.2 kJ/M of activation enthalpy and 0.35 kJ/mol K of activation entropy.

• Journal of Ginseng Research
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• v.22 no.1
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• pp.51-59
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• 1998

In this study, rat hepatocytes known to have active glucose metabolism were obtained to investigate the hypoglycemic action of fat soluble fraction of red ginseng by using the liver perfusion technique and incubated in two different media-one containing insulin and glucagon (control group), and the other containing glucagon only The activities of main regulating enzymes, such as glucokinase, glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenate, and glucose 6-phosphatase, related to metabolic pathways of glucose in these two kinds of hepatocytes were compared between these two groups and the effects of addition of fat soluble fraction ($10^1$~$10^4$%) from red ginseng to these two groups on these enzymes were also detected. The results were as follows. The specific activity of enzymes such as glucokinase, flucorse 6-phosphate dehydrogenase, and 6-phosphogluconate dehydrogenase related to glucose-consuming pathways of insulin-deficient group was much less than control one. However, their decreased activity was recovered after the addition of fat-soluble fraction at all range of concentrations. The specific activity of these enzymes after the addition of ginseng components to the control group was also increased. On the other hand, the specific activity of glucose 6-phosphatase related to glucose-producing pathway of insulin-deficient group was much higher than control one, but their increased activity was decreased obviously after the addition of fat soluble fraction at all range of concentrations. The same results were observed after the addition of fat-soluble fraction to the control group. These results suggest that the red ginseng saponin components might be effective on diabetic hyperglycemia by regulating the activity of enzymes related to glucose metabolism directly and/or indirectly. The effects of fat-soluble fraction ($10^2$%) and ginsenosides (mixture, $Rb_1$ and $Rg_1$, $10^4$%) on hypoglycemic action were compared. As a result, they showed considerable effect on hyperglycemia, but the best eff ect on the activities of glucokinase and glucose 6-phosphate dehydrogenase was appeared by ginsenoside $Rb_1$ and that of 6-phosphogluconate dehydrogenase and glucose 6-phosphatase was by ginsenoside mixture.

• Applied Biological Chemistry
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• v.40 no.5
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• pp.447-450
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• 1997

Eighteen N-substituted phenyl 1, 3, 5-trimethylpyrazole-4-carboxamides were synthesized to screen for their mycelial growth inhibition activity against Rhizoctonia solani $K{\ddot{u}}hn$ $(pEC_{50})$ and to measure enzymatic inhibition activity of these compounds $(pI_{50})$ against succinate dehydrogenase (SDH) isolated from Rhizoctonia solani $K{\ddot{u}}hn$ A structure-activity relationship formulated by regression analysis showed that 79% of the variance in mycelial growth inhibition activity can be explained with SDH inhibition activity and chromatographic capacity factor $(\acute{k})$ as a hydrophobic parameter related to the penetration and transport processes in the biological system.

• Journal of Microbiology
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• v.38 no.4
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• pp.275-280
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• 2000

Pseudomonas sp. S-47 is capable of catabolizing 4-chlorobenzoate (4CBA) as rarbon and energy sources under aerobic conditions via the mesa-cleavage pathway. 4CBA-dioxygenase and 4CBA-dihydrodiol dehydrogenase (4CBA-DD) catalyzed the degradation af 4CBA to produce 4-chlorocatechol in the pathway. In this study, the xylL gene encoding 4CBA-DD was cloned from the chromosomal DNA of Pseudomonas sp. S-47 and its nucleotide sequence was analyzed. The xylL gene was found to be composed of 777 nucleotide pairs and to encode a polypeptide of 28 kDa with 258 amino acid residues. The deduced amino acid sequence of the dehydrogenase (XylL) from strain S-47 exhibited 98% and 60% homologies with these of the corresponding enzymes, Pseudomonas putida mt-2 (XyIL) and Acinetobacter calcoaceticus (BenD), respectively. However, the amino arid sequences show 30% or less homology with those of Pseudomonas putida (BnzE), Pseudomonas putida Fl (TodD), Pseudomonas pseudoalcaligenes KF707 (BphB), and Pseudomonas sp. C18 (NahB). Therefore, the 4CBA-dihydrodiol dehdrogenase of strain S-47 belongs to the group I dehydrogenase involved in the degradation of mono-aryls with a carboxyl group.

• Current Research on Agriculture and Life Sciences
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• v.5
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• pp.68-74
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• 1987

The growth of A calcoaceticus C10 in CL medium was not increased by the addition of 0.5% E-capralactone or succinate, but increased by 0.2% adipate, xylose or even glucose which was not metabolized as a carbon source. The addition of 0.2% glucose after culture in CL medium for 6 hours increased the growth of A. colcoaceticus C10 twice as much as that in CL medium after culture for further 10 hours. Biosynthesis of cyclohexanol dehydrogenase in A. calcoaceticus C10 was not repressed by ${\varepsilon}$-caprolactone, succinate, xylose and glucose, but repressed by adipate which is endproduct of cyclohexanol metabolism. The induction of dehydrogenase by cyc1ohexanol in CL medium was not repressed completely by 0.1% adipate, but repressed almost completely by 0.2% adipate in A. calcoaceticus C10.

• Toxicological Research
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• v.35 no.1
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• pp.13-24
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• 2019

Numerous ethnomedicinal uses have been attributed to different parts of Annona senegalensis (ASE), including its uses as food and food additives. The present study investigated toxicological and antioxidant effects of 28 days administration of ethanol extracts of ASE stem bark to Wistar strain albino rats. Acute toxicity test was done to determine lethal dose in Wistar rats while sub-acute toxicity test was conducted on rats divided into four groups (A - control, B - 50 mg/kg, C - 100 mg/kg, D - 150 mg/kg, respectively and treated for 28 days. Oxidative stress markers in liver and kidney as well as hepatic succinate dehydrogenase activity in the mitochondrial and post mitochondrial fractions (PMF) were evaluated. The $LD_{50}$ value of ASE was > 2,000 mg/kg. White blood cell counts gradually increased, but red blood cell counts and haematocrits level decreased significantly (p < 0.05) by about 50%. Liver enzymes in the serum and mitochondrial succinate dehydrogenase activity increased significantly (p < 0.05). Superoxide dismutase and catalase activities also increased in liver mitochondria and PMF while malondialdehyde (MDA) and reduced glutathione levels increased only in the PMF. Furthermore, only MDA levels increased significantly in the kidney after 28 days extract administration. Histopathological examination showed hepatic necrosis and no obvious signs of nephrotoxicity. Anona senegalensis is relatively safe, but prolonged ingestion could induce oxidative stress and impair ATP synthesis through the modulation of the activity of mitochondrial succinate dehydrogenase.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.10
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• pp.1599-1605
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• 2005

The object of this study is to develop the method for differentiating fresh meat from frozen meat by using the measurement of the mitochondrial malate dehydrogenase in the Korean native cattle. The principle of this experiment is based on the fact that the enzyme proteins associated with mitochondrial membrane could be released by freezing. The methods of differentiating fresh meat from thawed, frozen meat were studied by measurements of mitochondrial malate dehydrogenase activity of meat press juice. Fresh and frozen beef were stored at 4, -4, -18 and -77$^{\circ}C$ for 15-day storage period. A meat press machine using air pressure was manufactured especially for these experiments, and sufficient amount of drip (about 0.15 mL/g) from 1.5 g of beef sample was efficiently obtained under a pressure of 8 kg/$cm^{2}$ generated by the meat pressing machine. The mitochondrial malate dehydrogenase activities of frozen meat drip i년ices stored at -18 and -77$^{\circ}C$ were significantly higher than those of fresh and frozen meat samples at -4$^{\circ}C$ (p < 0.05) during 10-min reaction period. However, the enzyme activities of the frozen meat drip juices (-18 and -77$^{\circ}C$) disappeared after 5 minutes of the reaction, which was not observed from the fresh and -4$^{\circ}C$ frozen meats. The enzyme activity maintained until 12 minutes for the fresh and -4$^{\circ}C$ frozen meats. From these results, the mitochondrial malate dehydrogenase could be considered as an indicator to differentiate fresh beef from frozen one.

• Journal of Microbiology and Biotechnology
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• v.6 no.2
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• pp.142-144
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• 1996

To clone the gene coding for steroid $\Delta^1$-dehydrogenase of Arthrobacter simplex, its genomic library was constructed with a , $\lambda$gt11 expression vector and immunoscreened with antiserum against the enzyme. One positive clone was found to carry a 1.6-kb EcoR I restriction endonuclease fragment of A. simplex DNA. The restriction map of the 1.6-kb EcoR I fragment was determined after cloning of the DNA into pBS vector.