• Journal of Microbiology and Biotechnology
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• v.26 no.12
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• pp.2076-2086
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• 2016

Fungal cytochrome P450 (CYP) enzymes catalyze versatile monooxygenase reactions and play a major role in fungal adaptations owing to their essential roles in the production avoid metabolites critical for pathogenesis, detoxification of xenobiotics, and exploitation avoid substrates. Although fungal CYP-dependent biotransformation for the selective oxidation avoid organic compounds in yeast system is advantageous, it often suffers from a shortage avoid intracellular NADPH. In this study, we aimed to investigate the use of bacterial glucose dehydrogenase (GDH) for the intracellular electron regeneration of fungal CYP monooxygenase in a yeast reconstituted system. The benzoate hydroxylase FoCYP53A19 and its homologous redox partner FoCPR from Fusarium oxysporum were co-expressed with the BsGDH from Bacillus subtilis in Saccharomyces cerevisiae for heterologous expression and biotransformations. We attempted to optimize several bottlenecks concerning the efficiency of fungal CYP-mediated whole-cell-biotransformation to enhance the conversion. The catalytic performance of the intracellular NADPH regeneration system facilitated the hydroxylation of benzoic acid to 4-hydroxybenzoic acid with high conversion in the resting-cell reaction. The FoCYP53A19+FoCPR+BsGDH reconstituted system produced 0.47 mM 4-hydroxybenzoic acid (94% conversion) in the resting-cell biotransformations performed in 50 mM phosphate buffer (pH 6.0) containing 0.5 mM benzoic acid and 0.25% glucose for 24 h at $30^{\circ}C$. The "coupled-enzyme" system can certainly improve the overall performance of NADPH-dependent whole-cell biotransformations in a yeast system.

• Journal of Microbiology and Biotechnology
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• v.23 no.6
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• pp.785-793
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• 2013

We studied the effects of 2 different dosages of high-molecular-weight poly-${\gamma}$-glutamic acid (hm ${\gamma}$-PGA) derived from Bacillus subtilis chungkookjang on lipid metabolism in a high-fructose diet-induced hypertriglyceridemic animal model. For 4 weeks, rats were fed either AIN-93 diet (normal control, NC; n = 10) or modified AIN-93 diet in which cornstarch was substituted with 63% fructose (n = 30) to induce hypertriglyceridemia. After 4 weeks, the hypertriglyceridemic rats were treated with daily oral doses of 0 mg (hypertriglyceridemic control, HC), 2.5 mg (hypertriglyceridemic, low hm ${\gamma}$-PGA, HL), or 5 $mg{\cdot}kg{\cdot}bw^{-1}{\cdot}d^{-1}$ (hypertriglyceridemic, high hm ${\gamma}$-PGA, HH) hm ${\gamma}$-PGA for 4 weeks. The HL and HH groups exhibited significantly lower levels of serum triglyceride, total cholesterol, LDL cholesterol, and free fatty acids than the HC group. The administration of hm ${\gamma}$-PGA reduced serum ALT and AST levels. The activities of lipogenic enzymes such as hepatic malic enzyme and glucose-6-phosphate dehydrogenase as well as glucose-6-phosphate dehydrogenase mRNA expression were significantly decreased by hm ${\gamma}$-PGA administration (p < 0.05). These results indicate that hm ${\gamma}$-PGA has an anti-hypertriglyceridemic effect in high-fructose diet-induced hypertriglyceridemic rats.

• Journal of Ginseng Research
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• v.17 no.1
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• pp.13-21
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• 1993

We attempted in this study to understand the hypoglycemic action of the fat soluble fraction of red ginseng roots in streptozotocin injected diabetic rats, through its actions on several enzymes relating to carbohydrate metabolism of the 1eve1 to compare with those of ginsenosides in streptozotocin injected diabetic rats. It was realized that the increased level of glucose, ketone bodies, lactate, nonesterified fatty acids and triacylglycerol in blood was significantly decreased and the decreased liver glycogen content of streptozotocin injected rats were appreciably moderated by intraperitoneal injection of the fat soluble fraction of red ginseng roots as shown in the saponin injected diabetic rats. The deceased activities of liver enzymes relating to carbohydrate metabolism such as phosphofructokinase, glucokinase, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and acetyl CoA carboxylase of streptozotocin induced diabetic rats were also sufficiently modified by the intraperitoneal injection of the above fat soluble fraction as shown in the ginsenoside injected streptozotocin induced rats.

• Korean Journal of Microbiology
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• v.14 no.4
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• pp.176-185
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• 1976

An improved procedure for the rapid purification of glucose-6-phosphate dehydrogenase from extracts of Saccharomyces cerevisiae was developed by using affinity chromatography. Among six affinty media tested, NADP$^{+}$-agarose and Affi-gel Blue were more effective than others (i.e., Affi-gel Red, AMP-agarose, ATP-agarose, and NAD$^{+}$-agarose). Conditions to desorb the enzyme bound to the affinity media were examined to increase the purity as well as yield. The best result was obtained when the column was developed with a linear gradient of KCl (0-1.0M). In case of Affi-gel Blue, introduction of NAD$^{+}$ (15mM) washing step prior to the salt gradient was most effective to remove NAD$^{+}$-binding proteins. For a large scale preparation of G-6-P dehydrogenase higher recovery was obtained by Affi-gel Blue than NADP$^{+}$-agarose, however, the purity of the enzyme was decreased by 10 times if the former was used as the affinity medium. The capacity of Affi-gel Blue for G-6-P dehydrogenase was found to be 5 times higher than that of NADP$^{+}$-agarose. Furthermore Affi-gel Blue could be reused repeatedly and its preparation is relatively easier and less expensive than NADP$^{+}$-agarose.X> +/-agarose.

• Reproductive and Developmental Biology
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• v.39 no.4
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• pp.97-104
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• 2015

Glucose is universal and essential fuel of energy metabolism and in the synthesis pathways of all mammalian cells. Glucose is the one of the major precursors of lactose synthesis using glycolysis result in producing milk fat and protein. During the milk fat synthesis, lipoprotein lipase (LPL) and CD36 are required for glucose uptake. Various morecules such as acyl-CoA synthetase 1 (ACSL1) activity of acetyl-CoA synthetase 2 (ACSS2), ACACA, FASN AGPAT6, GPAM, LPIN1 are closely related with milk fat synthesis. Additionally, glucose plays a major role for synthesizing lactose. Activations of lactose synthesize enzymes such as membranebound enzyme, beta-1,4-galactosyl transferase (B4GALT), glucose-6-phosphate dehydrogenase (G6PD) are changed by concentration of glucose in blood resulting change of amount of lactose production. Glucose transporters are a wide group of membrane proteins that facilitate the transport of glucose over a plasma membrane. There are 2 types of glucose transporters which consisted facilitative glucose transporters (GLUT); and sodium-dependent transport, mediated by the Na+/glucose cotransporters (SGLT). Among them, GLUT1, GLUT8, GLUT12, SGLT1, SGLT2 are main glucose transporters which involved in mammary gland development and milk synthesis. However, more studies are required for revealing clear mechanism and function of other unknown genes and transporters. Therefore, understanding of the mechanisms of glucose usage and its regulation in mammary gland is very essential for enhancing the glucose utilization in the mammary gland and improving dairy productivity and efficiency.

• Journal of Ginseng Research
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• v.13 no.1
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• pp.92-97
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• 1989

This study investigated the effects of high light intensity (100 KLw) and high temperature (45 ℃, dark) on enzyme (glucose-6-phosphate dehydrogenase, acid phosphatase, catalase, peroxidase, and proteinase) activities and characteristics of Panax ginseng C.A. Meyer leaves. Enzyme activity and protein content decreased rapidly under treatment with high light intensity In P ginseng the thermal stabilities of catalase and peroxidase were high (above 70%), and the coagulation rates of soluble proteins were low (below 17%). Therefore, the decrease in enzyme activity and protein content was not caused by increase in leaf temperature due to the high light intensity, but by increase in proteolytic activities. The photochemical formation rate of superoxide radical (O-2) was higher in the P ginseng leaf extracts than in Solanum nigmm, and was accelerated by addition of crude saponin to the buffer extracts.

• Journal of Nutrition and Health
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• v.30 no.4
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• pp.386-393
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• 1997

This experiment was performed to investigate the effect of different dietary lipids on the risk factors of coronary vascular disease(CVD) in ovariectomized rats. female rats of Sprague-Dawley stram were divided into sham-operated(sham) and ovariectomized(ovx) groups and then each group was divided into a beef fallow group, a soy bean oil group and a fish oil group. After 16 weeks of feeding on experimental diets, animals were sacrificed and blood, liver, kidney and perirenal fat pad were obtained. Food intake and weight gain of fish oil group were significantly lower than other dietary lipid groups. food intake and weight gain tended to be higher in ovx groups than in sham groups. The weight Index(g/100g body weight) of liver and kidney was higher in the fish oil group than the other groups and weight index was lower in ovx groups compared to sham groups. The weight of the perirenal fat pad was the highest in the beef tallow group and the lowest in the fish oil group. The fish oil group showed the lowest total cholesterol(TC) and triglyceride (TG) levels in serum. Serum TG levels were lower in all ovx groups than in sham groups, but serum TC levels were not influenced by ovariectomy. fatty acid composition of serum reflects the recent dietary Intake of fat. Linoleic acid content was tile highest in soy bean oil group and eicosapentaenoic acid(EPA) and docosahexaenoic acid(DHA) contents were the highest in fish oil group. fatty acid composition of adipose tissue, especially EPA and DHA contents in perirenal fat pad, was highest in the fish oil group. Saturated fatty acid(SFA) and monounsaturated fatty acid(MUFA) in serum and adipose tissue did not reflect fatty acid intake. The activities of glucose-6-phosphate dehydrogenase, a lipogenic enzyme, in the blood of the beef tallow and soybean oil groups showed the tendency to be high and that of the fish oil group to be low in ovx. Carnitine acetyltransferase, a lipolytic enzyme, showed the highest activity in the liver of the fish oil group and was least active in the soy bean oil group.

• Journal of the Korean Applied Science and Technology
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• v.30 no.3
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• pp.394-399
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• 2013

This study was performed to investigate the antidiabetic study effect in ethanol extract of Ailanthicortex(A.C) radici in Streptozotocin(STZ)-induced diabetic rats. Diabetes was induced by intravenous injection of STZ at a dose of 45mg/kg dissolved in citrate buffer. The ethanol extract of A.C radici was orally administrated once a day for 7 days at a dose of 500mg/kg. The content of serum glucose, triglyceride(T.G), total cholesterol were significantly decreased in treated group compared to the those of STZ-control group. The content of hepatic glycogen and activity of glucokinase(GK) were significantly increased in treated group compared to the those of STZ-control group. but activity of glucose-6-phosphate dehydrogenase(G-6-PDH) was significantly increased. These results indicated that ethanol extract of A.C would have antidiabetic effect in STZ-induced diabetic rats.

• Food Science and Biotechnology
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• v.17 no.2
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• pp.425-429
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• 2008

The anti-obesity effect of chokong, Rhynchosia nolubilis seeds pickled in vinegars for 2 weeks at $4^{\circ}C$, was investigated. During the differentiation of 3T3-L1 adipocytes, the addition of ethanolic extracts of chokongs lowered the cellular triglyceride content by 8.1-9.0%, and glucose content by 12.2-27.6%, depending on the kinds of vinegar used. The activity of glycerol-3-phosphate dehydrogenase also decreased up to 56.0-59.3% by supplying those extracts. In addition, vinegars were superior to acetic acid, citric acid, and hydrochloric acid solutions, and distilled water in anti-obesity of the pickled seeds.

• Journal of Ginseng Research
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• v.7 no.1
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• pp.68-73
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• 1983

E. coli (K-12 W1485) was grown in M9 minimal medium containing ginseng saponin (10-3%-2%) and found that the cells grew most rapidly in the presence of 10-1% saponin. The cells, harvested at the early exponential phase, were transferred to the minimal medium containing 10-1% saponin plus 14C-labelled saponin (0.03 ${\mu}$Ci) and the incubation was continued at 37$^{\circ}C$ for 20 minutes and the cells were fractionated into the outermembrane, innermembrane and cytosol fraction. Radioactivity data showed that the most radioactivity was detected in the innermembrane. The activity of succinate dehydrogenase of the cells grown in the above saponin medium was significantly higher than that of the cells grown in ordinary minimal medium. No significant difference of the glucose 6-phosphate dehydrogenase activity was observed between the two groups. It was also found that the saponin stimulated glucose uptake and biosynthisis of lipids and proteins of the cells. Incorporation of 14C-leucine into the protein fraction of the cell was also accelerated.