Ibraheem, Omodele;Adewale, Isaac Olusanjo;Afolayan, Adeyinka
BMB Reports
/
v.38
no.5
/
pp.584-590
/
2005
Glucose 6-phosphate dehydrogenase (EC 1.1.1.49) was purified from Aspergillus aculeatus, a filamentous fungus previously isolated from infected tongue of a patient. The enzyme, apparently homogeneous, had a specific activity of $220\;units\;mg^{-1}$/, a molecular weight of $105,000{\pm}5,000$ Dal by gel filtration and subunit size of $52,000{\pm}1,100$ Dal by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The substrate specificity was extremely strict, with glucose 6-phosphate (G6P) being oxidized by nicotinamide adenine dinucleotide phosphate (NADP) only. At assay pH of 7.5, the enzyme had $K_m$ values of $6\;{\mu}m$ and $75\;{\mu}m$ for NADP and G6P respectively. The $k_{cat}$ was $83\;s^{-1}$. Steady-state kinetics at pH 7.5 produced converging linear Lineweaver-Burk plots as expected for ternary-complex mechanism. The patterns of product and dead-end inhibition suggested that the enzyme can bind NADP and G6P separately to form a binary complex, indicating a random-order mechanism. The enzyme was irreversibly inactivated by heat in a linear fashion, with G6P providing a degree of protection. Phosphoenolpyruvate (PEP), adenosinetriphosphate (ATP), and fructose 6-phosphate (F6P), in decreasing order, are effective inhibitors. Zinc and Cobalt ions were effective inhibitors although cobalt ion was more potent; the two divalent metals were competitive inhibitors with respect to G6P, with $K_i$ values of $6.6\;{\mu}m$ and $4.7\;{\mu}m$ respectively. It is proposed that inhibition by divalent metal ions, at low NADPH /NADP ratio, is another means of controlling pentosephosphate pathway.
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.
For mass production of poly(3-hydroxybutyrate) (PHB), high cell density cultures of Ralstonia eutropha were carried out in 2.5-1 and 60-1 fermentors by two fed-batch culture techniques of nitrogen and phosphate limitation. When the nitrogen limitation technique was employed using both an on-line glucose monitoring and control system, a high concentration level of PHB (121g/l) was obtained in the small-scale fermentor of 2.5 1. However, the PHB concentration obtained in a large-scale fermentor of 60 1 only turned out to be 60g/l. In contrast, when another fed-batch culture technique of the phosphate-limitation employing dissolved oxygen (DO) stat glucose feeding was used, a large amount of PHB was successfully produced in both 60-1 and 2.5-1 fermentors. In a 2.5-1 fermentor, concentrations of PHB and cells obtained in 58 h were 175 and 210 g/l, respectively, which corresponded to the PHB productivity level of 3.02 g/l/h. In a 60-1 fermentor, a final cell concentration of 221 g/l and a PHB concentration of 180 g/l with PHB productivity level of 3.75 g/l/h were obtained in 48h. PHB content and yield from glucose were 81% and 0.38g PHB/g glucose, respectively. These data suggest that the phosphate limitation technique is more effective compared to nitrogen limitation in the mass production of PHB by R. eutropha of a large scale.
Cho, Eunae Sandra;Cha, Yong Hoon;Kim, Hyun Sil;Kim, Nam Hee;Yook, Jong In
Biomolecules & Therapeutics
/
v.26
no.1
/
pp.29-38
/
2018
During cancer progression, cancer cells are repeatedly exposed to metabolic stress conditions in a resource-limited environment which they must escape. Increasing evidence indicates the importance of nicotinamide adenine dinucleotide phosphate (NADPH) homeostasis in the survival of cancer cells under metabolic stress conditions, such as metabolic resource limitation and therapeutic intervention. NADPH is essential for scavenging of reactive oxygen species (ROS) mainly derived from oxidative phosphorylation required for ATP generation. Thus, metabolic reprogramming of NADPH homeostasis is an important step in cancer progression as well as in combinational therapeutic approaches. In mammalian, the pentose phosphate pathway (PPP) and one-carbon metabolism are major sources of NADPH production. In this review, we focus on the importance of glucose flux control towards PPP regulated by oncogenic pathways and the potential therein for metabolic targeting as a cancer therapy. We also summarize the role of Snail (Snai1), an important regulator of the epithelial mesenchymal transition (EMT), in controlling glucose flux towards PPP and thus potentiating cancer cell survival under oxidative and metabolic stress.
Journal of the Korean Applied Science and Technology
/
v.37
no.4
/
pp.682-687
/
2020
This study was performed to investigate the antidiabetic effect of ethanol extract of Phelladrindron Amurense Rupr (P.A) in Streptozotocin(STZ) induced diabetic rats. Diabetes was induced by intravenous injection of STZ at a dose of 45mg/kg,b.w. dissolved in citrate buffer. The ethanol extract of P. A was orally administrated once a day for 7 days at a dose of 1,000mg/kg. The content of serum glucose, was significantly decreased in P.A treated group compared to the those of STZ-control group. The content of hepatic glycogen and activities of glucokinase(GK) and glucose-6-phosphate dehydrogenase(G-6-PDH) were significantly increased(p<0.05), and activity of glucose-6-phoshatase(G-6-Pase) was significantly decreased(p<0.05) in P.A treated group compared to those of STZ-control group, These results indicated that ethanol extract of P.A have antidiabetic effect in STZ-induced diabetic rats.
Kim, Young-Ok;Park, In-Suk;Nam, Bo-Hye;Kong, Hee-Jeong;Kim, Woo-Jin;Lee, Sang-Jun;Kim, Kyung-Kil
Fisheries and Aquatic Sciences
/
v.13
no.1
/
pp.49-55
/
2010
A bacterial strain with phytase and glucose-1-phosphatase activity was isolated from seawater. The colony was identified as an Enterobacter cloacae strain and named E. cloacae B11. A gene, agpEnB11, coding for an intracellular acid glucose phosphatase was cloned from the strain and sequenced. It comprised 1,242 nucleotides and encoded a polypeptide of 413 amino acids. Recombinant glucose-1-phosphatase (AgpEn) was overexpressed in Escherichia coli and purified using Ni-NTA column under native conditions. Purified protein displayed a single band of 47 kDa on SDS-PAGE. AgpEn hydrolyzed a wide variety of phosphorylated compounds, with high activity for glucose-1-phosphate and glucose-6-phosphate. Optimum pH and temperature for enzyme activity were pH 5.0 and $50^{\circ}C$, respectively. Enzyme activity was stimulated by $Ca^{2+}$ and $Co^{2+}$, and inhibited by $Cu^{2+}$.
Kim, Ik-Sang;Hong, Tae-Yee;Lee, Woo-Kon;Chang, Woo-Hyun
The Journal of the Korean Society for Microbiology
/
v.20
no.1
/
pp.55-63
/
1985
Phosphate, ammonia, glucosamine, glucose, pyruvate, succinate, fumarate, malate and acetate were examined for their ability to control the heat-stable enterotoxin (ST) production in succinate salts medium or in M9 medium. The results obtained were summerized as follows. 1. When the initial phosphate concentration was adjusted to 1.0mM, ST production was decreased to 80u/ml or less. But when the initial phosphate concentration was adjusted to 64mM or 100mM, enterotoxin production was 320u/ml. 2. When the initial ammonia concentration in the medium was adjusted to 1.0mM, no ST production and cell growth were observed. But when ammonia concentration was adjusted to 10mM, 19mM, 38mM or 76mM, enterotoxin production was 320u/ml. 3. Among carbon sources, glucosamine, glucose, pyruvate, succinate, fumarate, malate and acetate, acetate supported the highest specific production (928 unit/O.D.) of heat-stable enterotoxin. From this results, we could assume that heat-stable enterotoxin production is controlled by stringent control mechanism. 4. When the pH of the succinate salts medium was kept between 6.2 to 6.5, no heat-stable enterotoxin production was observed, but when the pH of the medium was kept between pH 6.2 to 6.5, 267 unit/O.D. of heat-stable enterotoxin was produced. 5. Glucose inhibited the heat-stable enterotoxin production and the mechanism was assumed due to its capacity to lower the pH of the medium during catabolysis and its high metabolic energy.
Nucleic acid and protein biosynthese of the glucose-bleached Chlorella cells in relation to the process of the chloroplast reformation were traced, by measuring the changes in the amounts of cell constituents and nuclease activities of the cells during the greening process. The contents of RNA and protein of the glucose-bleached cells decreased significantly, shile the contents of nucleotides and amino acids of the cells increased to compared with those of the control, showing that the biosynthetic activities of RNA and protein of the cells were inhibited severely in the glucose-bleaching process. In the early greening process of the glucose-bleached Chlorella cells the contents of RNA and protein of the cells increased significantly, while the contents of nucleotides nad amino acids of the cells increased to compared with those of the control, showing that the biosynthetic activities of RNA and protein of the cells were inhibited severely in the glucose-bleaching process. In the early greening process of the glucose-bleached Chlorella cells the contents of RNA and protein of the cells increased significantly wihout any increase in the chlorophyll contents showing that the massive biosynthese of RNA and protein proceed prior to the chlorophyll bioynthesis in the cells. The phosphate contents in the DNA fraction of the glucose-bleached cells decreased, but the contents of acid-insoluble polyphosphate increased to compared with those of the control in the early greening porcess, exhibiting that the incorporation of the phosphorus from acid-insoluble polyphosphate into DNA was retarded. In the greening process of the glucose-bleached cells the ribonuclease nad deoxyribonuclease activities of the cells decreased to compared with those of the control, although the initial activities of the both enzymes in the cell were far great compared with the control. Although the initial phosphate contents in the lipid fraction of the glucose-bleached Chlorella cells were more great than the control, the phosphate contents in the lipid fraction of the cells decreased in the early greening process to compared with control, and then increased in the late developmental stages in which massive chlorophyll biosynthesis occured.
RhodopseudolllOllas palustris P4 was studied for $H_2$ production from glucose in batch culture. Important conditions studied include phosphate concentration, initial pH, temperature, glucose concentration, and gas-phase replacement. Optimal $H_2$ production was observed at 60 - 300 mM of phosphate and 7.8 - 8.6 of initial pH. The effect of culture temperature was negligible When glucose concentration increased from 0.1 to 5% (w/v), $H_2$ production increased up to 2% and remained constant thereafter. Intermittent purging of the reaction botlle with Ar gas stimulated the Hl production by alleviating the inhibition by $H_2$. The maximum productivity was 111.1 ml $H_2$/h-1.
In this study Bacillus subtilis PTCC 1023 was used for the production of protopectinase using soybean based media. The use of isolated soybean protein (ISP) and soybean flour resulted in similar protopectinase production and growth rates. The effect of medium composition on protopectinase production was studied using central composite design (CCD) methodology. The change in the concentration of ISP (1-7%), glucose (0-10%), and phosphate (0.1-0.3 M) was found to affect the protopectinase activity (response variable) after 24 hr of cultivation. In the range studied, ISP and glucose had a negative effect on the response variable, whereas phosphate had a positive effect. A statistically significant interaction was identified between phosphate and ISP, suggesting that correct optimization of medium formulation in this case can only be obtained using factorial design of experiments. Protopectinase activity exceeding 215 U/mL was obtained in a medium containing 4% ISP, 0.3M phosphate, and no added sugar.
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