• Title/Summary/Keyword: Glucose-1-phosphate

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Antidiabetic effect of ethanol extract on Codonopsis lanceolata root in streptozotocin induced diabetic rats (더덕 뿌리 에탄올 추출물이 streptozotocin으로 유발된 흰쥐의 항 당뇨효과)

  • Kim, Ok-Kyung
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.2
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    • pp.271-277
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    • 2016
  • This study was done to investigate the antidiabetic effect of ethanol extract from Codonopsis lanceolata root in Streptozotocin (STZ)-induced diabetic rats. Diabetes was induced by intravenous injection of STZ at a dose 45mg/kg.b.w. dissolved in citrate buffer(pH4.5). The ethanol extract of Codonopsis lanceolata root was orally administrated once a day for 7 days. The contents of serum glucose, triglyceride(TG) and total cholesterol were significantly decreased(p<0.05) in Codonopsis lanceolata root treated group compared to the those of STZ-control group. Also the content of hepatic glycogen and activities of glucose-phosphate dehydrogenase(G-6-PDH) and glucokinase(GK) were significamtly increased(p<0.05). These results indicated that ethanol extract of Codonopsis lanceolata root would have antidiabetic effect in STZ-induced diabetic rats.

Cloning, Expression, and Characterization of UDP-glucose Pyrophosphorylase from Sphingomonas chungbukensis DJ77

  • Yoon, Moon-Young;Lee, Kyoung-Jin;Park, Hea-Chul;Park, Sung-Ha;Kim, Sang-Gon;Kim, Sung-Kun;Choi, Jung-Do
    • Bulletin of the Korean Chemical Society
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    • v.30 no.6
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    • pp.1360-1364
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    • 2009
  • The bacterium Sphingomonas chungbukensis DJ77 produces the extracellular polysaccharide gellan in high yield. Gellan produced by this bacterium is widely used as a gelling agent, and the enzyme UDP-glucose pyrophosphorylase (UGP) is thought to play a key role in the gellan biosynthetic pathway. The UGP gene has been successfully cloned and over-expressed in E. coli. The expressed enzyme was purified with a molecular weight of approximately 32 kDa, as determined by a SDS-polyacrylamide gel, but the enzyme appears as ca. 63 kDa on a native gel, suggesting that the enzyme is present in a homodimer. Kinetic analysis of UDP-glucose for UGP indicates $K_m$ = 1.14 mM and $V_{max}$ = 10.09 mM/min/mg at pH 8.0, which was determined to be the optimal pH for UGP catalytic activity. Amino acid sequence alignment against other bacteria suggests that the UGP contains two conserved domains: An activator binding site and a glucose-1-phosphate binding site. Site-directed mutagenesis of Lys194, located within the glucose-1-phosphate binding site, indicates that substitution of the charge-reversible residue Asp for Lys194 dramatically impairs the UGP activity, supporting the hypothesis that Lys194 plays a critical role in the catalysis.

Cloning and Idendification of dTDP-L-Rhamnose Biosynthetic Gene Cluster from Thermus caldophilus GK24

  • Kim, Ki-Chan;Lee, Seung-Don;Han, Ju-Hee;Sohng, Jae-Kyung;Liou, Kwang-Kyoung
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.749-754
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    • 2000
  • PCR primers were designed based on consensus sequences of dTDP-D-glucose 4,6-dehydratase, one of the enzymes involved in the biosynthesis of deoxysugar. The PCR product (360 bp) was obtained from Thermus caldophilus GK24. Colony hybridization was carried out to the cosmid library constructed from T. caldophilus GK24 genomic DNA by the PCR product DNA fragment. We isolated a cosmid clone (pSMTC-1) that was subcloned to call pKCB series plasmid (BamHI fragments), partially sequenced and analyzed. pKCB80 (4.2 kb-BamHI DNA fragment) of them showed ORFs that was orfA, orfB, orfC and orfD. The orfABCD gene cluster is the deosysugar biosynthetic gene ; orfA (glucose-1-phosphate thymidylytransferase), orfB (dTDP-D-glucose 4,6-dehydratase), orfC (dTDP-4-keto-L-rhamnose reductase) and orfD (dTDP-4-keto-6-deoxy-D-glucose 3,5-epimerase). The gene cluster that was related in biosynthesis of dTDP-L-rhamnose was also identified by computer analysis, and we proposed that the biosynthetic pathway of deoxysugar analyzed from DNA sequencing of pKCB80 is from D-glucose-1-phosphate, dTDP-D-glucose, dTDP-4-keto-6-deoxy-D-glucose via dTDP-4-keto-L-rhamnose to dTDP-L-rhamnose.

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Physiological studies on cell division by the technique of synchronous culture of chlorella (I) (클로렐라의 동조배양법에 의한 세포분열의 생리학적 연구 1)

  • 이영녹;이종삼
    • Korean Journal of Microbiology
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    • v.7 no.1
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    • pp.1-9
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    • 1969
  • Changes in the phosphorylation of Chlorella cells during the life cycle the aulotrophic and micotrophic synchronous culture were followed under the light and dark. 1. In the autotrophic culture of Chlorella the amounts of esterified phosphate compounds of the algal cell under the light increased during the growing period and decreased strikingly in the ripening period showing a peak at the $L_1$ i/-cell stage. 2. TRhe amount of total esterified phosphate compounds of the cell under the dark, however, decreased during the growing period and then kept fairly constnat during the ripening nad division periods showing the greates activity of the oxidative phosphorylation in the early growing stage. 3. It is presumed that the energy requirement of the dividing algal cell in the autotrophic culture is fulfilled prior to the nuclear division mostly by the photosynthetic phosphorylation. 4. In the mixotrophic culture, the amount of esterified phosphate compounds of the algal cells under the light increased during the growing period and decreased during the late ripening and early division periods showing a peak in the $L_2$-cell stage as in the case of the phosphorylation under the dark. 5. The phosphorylation of the fell grown in the glucose medium is more active under the dark than under the light in the stages of the growing and early ripening periods. 6. It is considered that the excess glucose in the algal cell not only promotes the oxidative phosphorylation but also inhibits the photosynthetic phosphorylation of the cell. 7. It is presumed that the energy requirement of the dividing algal cell in the glucose medium is fulfilled prior to the nuclear division by the combined action of oxidative and photosynthetic phosphorylation, mostly by the oxidative phosphorylation.

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Media Optimization of Corynebacterium glutamicum for Succinate Production Under Oxygen-Deprived Condition

  • Jeon, Jong-Min;Thangamani, Rajesh;Song, Eunjung;Lee, Hyuk-Won;Lee, Hong-Weon;Yang, Yung-Hun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.2
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    • pp.211-217
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    • 2013
  • Corynebacterium glutamicum is one of the well-studied industrial strain that is used for the production of nucleotides and amino acids. Recently, it has also been studied as a possible producer of organic acids such as succinic acid, based on its ability to produce organic acids under an oxygen deprivation condition. In this study, we conducted the optimization of medium components for improved succinate production from C. glutamicum under an oxygen deprivation condition by Plackett-Burman design and applied a response surface methodology. A Plackett-Burman design for ten factors such as glucose, ammonium sulfate, magnesium sulfate, potassium phosphate ($K_2HPO_4$ and $KH_2PO_4$), iron sulfate, manganese sulfate, biotin, thiamine, and sodium bicarbonate was applied to evaluate the effects on succinate production. Glucose, ammonium sulfate, magnesium sulfate, and dipotassium phosphate were found to have significant influence on succinate production, and the optimal concentrations of these four factors were sequentially investigated by the response surface methodology using a Box-Behnken design. The optimal medium components obtained for achieving maximum concentration of succinic acid were as follows: glucose 10 g/l, magnesium sulfate 0.5 g/l, dipotassium phosphate ($K_2HPO_4$) 0.75 g/l, potassium dihydrogen phosphate ($KH_2PO_4$) 0.5 g/l, iron sulfate 6 mg/l, manganese sulfate 4.2 mg/l, biotin 0.2 mg/l, thiamine 0.2 mg/l, and sodium bicarbonate 100 mM. The parameters that differed from a normal BT medium were glucose changed from 40 g/l to 10 g/l, dipotassium phosphate ($K_2HPO_4$) 0.5 g/l changed to 0.75 g/l, and ammonium sulfate ($(NH_4)_2SO_4$) 7 g/l changed to 0 g/l. Under these conditions, the final succinic acid concentration was 16.3 mM, which is about 1.46 fold higher than the original medium (11.1 mM) at 24 h. This work showed the improvement of succinate production by a simple change of media components deduced from sequential optimization.

Biosynthetic Regulation and Enzymatic Properties of $\beta$-Glucosidase from Cellulomonas sp. CS 1-1 (Cellulomonas sp. CS1-1으로 부터의 $\beta$-Glucosidase의 합성조절과 그의 효소학적 성질)

  • Lee, Hee-Soon;Min, Kyung-Hee;Bae, Moo
    • Microbiology and Biotechnology Letters
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    • v.16 no.2
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    • pp.119-125
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    • 1988
  • $\beta$-Glucosidase of Cellulomonas sp. CS1-1 in cellular compartment was localized with cell-bound form while Avicelase and carboxymethylcellulase (CMCase) were appeared with extracellular enzyme. Cell growth on cellulose or CMC minimal broth was increased by glucose addition. $\beta$-Glucosidase production on cellobiose or CMC minimal broth was repressed by the addition of glucose. However, on CMC minimal broth, the enzyme production was specially stimulated by cellobiose addition. $\beta$-Glucosidase production was also induced by CMC, starcth and maltose compared with glycerol, arabinose, xylose and trehalose. From the above results, it was concluded that glucose effect on $\beta$-glucosidase biosynthesis showed catabolite repression, but enzyme production was induced by cellobiose, CMC, and starch, indicating that $\beta$-glucosidase is inducible enzyme. Yeast extract stimulated $\beta$-glucosidase production more than peptone and ammonium sulfate. $\beta$-Glucosidase activity was increased with 50mM MgCl$_2$in 10mM potassium phosphate buffer (pH 7.0). Optimum conditions for enzyme activities were pH 6.0 and 42$^{\circ}C$, Km value of $\beta$-glucosidase for p-nitrophenyl-$\beta$-D-glucosidase was 0.256mM and Ki for $\beta$-D(+)-glucose was 9.0mM.

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The Effect of Glucose and Glucose Transporter on Regulation of Lactation in Dairy Cow

  • Heo, Young-Tae;Park, Joung-Jun;Song, Hyuk
    • 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.

Short-Cut Pathway to Synthesize Cellulose of Encysting Acanthamoeba

  • Moon, Eun-Kyung;Kong, Hyun-Hee
    • Parasites, Hosts and Diseases
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    • v.50 no.4
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    • pp.361-364
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    • 2012
  • The mature cyst of Acanthamoeba is highly resistant to various antibiotics and therapeutic agents. Cyst wall of Acanthamoeba are composed of cellulose, acid-resistant proteins, lipids, and unidentified materials. Because cellulose is one of the primary components of the inner cyst wall, cellulose synthesis is essential to the process of cyst formation in Acanthamoeba. In this study, we hypothesized the key and short-step process in synthesis of cellulose from glycogen in encysting Acanthamoeba castellanii, and confirmed it by comparing the expression pattern of enzymes involving glycogenolysis and cellulose synthesis. The genes of 3 enzymes, glycogen phosphorylase, UDP-glucose pyrophosphorylase, and cellulose synthase, which are involved in the cellulose synthesis, were expressed high at the 1st and 2nd day of encystation. However, the phosphoglucomutase that facilitates the interconversion of glucose 1-phosphate and glucose 6-phosphate expressed low during encystation. This report identified the short-cut pathway of cellulose synthesis required for construction of the cyst wall during the encystation process in Acanthamoeba. This study provides important information to understand cyst wall formation in encysting Acanthamoeba.

The Optimum Culture Condition for the Increasement of Biosurfactant Produced by Pseudomonas aeruginosa F722 (생물계면활성제 생산증가를 위한 Pseudomonas aeruginosa F722의 최적배양조건)

  • 오경택;강창민;정선용
    • KSBB Journal
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    • v.18 no.2
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    • pp.145-148
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    • 2003
  • The hydrogen-producting bacterium was isolated from fresh water and identified as Enterbacter cloacae. The isolated was named Enterobacter cloacae YJ-1. In batch culture, The optimum cultivation temperature and pH of strain YJ-1 was 35$^{\circ}C$ and 7.5, respectively. All of the added glucose was consumed completely during fermentation even though pH was not controlled. Amount of hydrogen produced on each condition of 2% glucose, 4% sucrose and 5% fructose was 950, 1000 and 948 mL/L, respectively and resulted in increasing hydrogen production approximately 2.5-times more than controlled condition. The macimum hydrogen production was obtained when 50mM phosphate was added. was obtained when 50mM phosphate was added. In repeated0batch culture, yeast extract, but the production amount was not changed on the condition of over 0.5%, Most of the organic acides produced during the fermentation were formic and acetic acid, and propionic acid was moiety also generated.

Phosphate Solubilizing Activity of Pseudomonas sp. CL-1 and Kluyvera sp. CL-2 (Pseudomonas sp. CL-1 및 Kluyvera sp. CL-2 균주의 인산가용화 특성)

  • Kwon, Jang-Sik;Suh, Jang-Sun;Weon, Hang-Yeon;Kim, Wan-Gyu;Noh, Hyung-Jun
    • Korean Journal of Soil Science and Fertilizer
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    • v.40 no.6
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    • pp.442-446
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    • 2007
  • From the rhizoplane and rhizosphere of pepper, tomato, lettuce, pasture, and grass, unsoluble inorganic phosphate solubilizing bacterial strains were isolated using plate base assay on Pikovskaya's medium. Two strains, CL-1 and CL-2, which produced largest halo on plates (indicative of phosphate solubilization)were selected for further studies. Based on these biochemical and 16S rRNA analysis strains CL-1, CL-2 were found to be as species of Pseudomonas sp. and Kluyvera sp., respectively. In broth assay Pseudomonas sp. CL-1 and Kluyvera sp. CL-2 solubilized insoluble phosphate by 193.4 mg and $493.6P\;mg\;L^{-1}$, respectively after $3^{rd}$ day inoculation. These effecient phosphate solubilizing bacteria have a potential to be developed as microbial based fertilizer in future.