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

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Characterization of the $\alpha$-Galactosidase Gene from Leuconostoc mesenteroides SY1

  • KIM JONG HWAN;PARK JAE-YONG;JEONG SEON-JU;CHUN JIYEON;LEE JONG HOON;CHUNGZ DAE KYUN;KIM JEONG HWAN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.800-808
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    • 2005
  • Leuconostoc mesenteroides SY1, an isolate from kimchi, was able to ferment $\alpha$-galactosides, such as melibiose and raffinose. $\alpha$-Galactosidase ($\alpha$-Gal) activity was higher in cells grown on melibiose and raffinose than cells grown on galactose, sucrose, and fructose. $\alpha$-Gal activity was not detected in cells grown on glucose, indicating the operation of carbon catabolite repression (CCR). A 6 kb DNA fragment was PCR amplified using a primer set based on the nucleotide sequence of a putative $\alpha$-galactosidase gene (aga) from L. mesenteroides ATCC 8293. Nucleotide sequencing of the 6 kb fragment confirmed the presence of aga and other genes involved in the galactosides utilization, and the gene order was galR (transcriptional regulator)-aga-gaIK (galactokinase)-gaIT (galactose-1-phosphate uridylyltransferase). Northern blotting experiment showed that aga, gaIK, and gaIT constituted the same operon, that the transcription was induced by galactosides, such as melibiose and raffinose, whereas gaIR was independently transcribed as a monocistronic gene, and that the level of transcription was fairly constant. The aga was overexpressed in E. coli BL21 (DE3) using pET26b(+) vector, and $\alpha$-Gal was accumulated in E. coli as an inclusion body.

Proteome Analysis of Paenibacillus polymyxa E681 Affected by Barley

  • Seul, Keyung-Jo;Park, Seung-Hwan;Ryu, Choong-Min;Lee, Yong-Hyun;Ghim, Sa-Youl
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.934-944
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    • 2007
  • Paenibacillus polymyxa E681 is known to be able to suppress plant diseases by producing antimicrobial compounds and to promote plant growth by producing phytohormones, and secreting diverse degrading enzymes. In spite of these capabilities, little is known regarding the flow of information from the bacterial strain to the barley roots. In an attempt to determine the flow of information from the bacterial strain to barley roots, the strain was grown in the presence and absence of barley, and two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and MALDI-TOF mass spectrometry were used. 2D-PAGE detected approximately 1,000 spots in the cell and 1,100 spots in the supernatant at a pH 4-10 gradient. Interestingly, about 80 spots from each sample showed quantitative variations. Fifty-three spots from these were analyzed by MALDI-TOF mass spectrometry and 28 proteins were identified. Most of the cytosolic proteins expressed at higher levels were found in P. polymyxa E681 cells grown in the presence of barley rather than in the absence of barley. Proteins detected at a lower level in the surpernatant of P. polymyxa E68l cells grown in the presence of barley were lipoprotein, glucose-6-phosphate 1-dehydrogenase, heat-shock protein HtpG, spermidine synthase, OrfZ, ribonuclease PH, and coenzyme PQQ synthesis protein, and flagellar hook-associated protein 2 whereas proteins detected at a higher level in the surpernatant of P. polymyxa E681 cells grown in the presence of barley included D-alanyl-D-alanine ligase A, isopentenyl-diphosphate delta-isomerase, ABC transporter ATP-binding protein Uup, lipase. Many of the proteins belonging to plant-induced stimulons are associated with biosynthetic metabolism and metabolites of proteins and transport. Some of these proteins would be expected to be induced by environmental changes resulting from the accumulation of plant-secreted substances.

Effects of Osmolarity and Vitamins on the In Vitro Development of Bovine Embryos in a Chemically Defined, Protein-free Culture Medium (무단백 한정배양액에서 삼투압 및 비타민이 소 수정란의 체외발생에 미치는 영향)

  • 김종홍;이상찬;김병기
    • Korean Journal of Animal Reproduction
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    • v.20 no.1
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    • pp.77-84
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    • 1996
  • The purpose of this study was to evaluate effects of osmolarity and vitamins on the in vitro development of embryos. Bovine embryo that had been matured and fertilized in vitro were cultured in simple, chemically defined, protein free medium (mTLP-PVA). When the osmolarity of the medium supplemented 0.35 mM phos-phate and 19 amino acid was changed by NaCI concentration, significantly(p<0.05) higher pro-portion of 1-cell embryos in the medium of 265 or 290 mOsm developed to the morula (32 ~ 35%) and blastocyst(24 ~ 28%) stage. When embryos were transferred to fresh medium containing 5.56mM glucose at 120hrs post-insemination, the highest proportion of embryos developed to mor-ula(40%) and blastocyst(32%) stages at 290 mOsm(p<0.05), although the value in morulae was not significantly different with that(35%) at 315 mOsm. Vitamins in presence of glutamine and amino acids had no beneficial effects on the development of 1-cell embryos to the blastocyst.

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Dietary ε-Polylysine Decreased Serum and Liver Lipid Contents by Enhancing Fecal Lipid Excretion Irrespective of Increased Hepatic Fatty Acid Biosynthesis-Related Enzymes Activities in Rats

  • Hosomi, Ryota;Yamamoto, Daiki;Otsuka, Ren;Nishiyama, Toshimasa;Yoshida, Munehiro;Fukunaga, Kenji
    • Preventive Nutrition and Food Science
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    • v.20 no.1
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    • pp.43-51
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    • 2015
  • ${\varepsilon}$-Polylysine (EPL) is used as a natural preservative in food. However, few studies have been conducted to assess the beneficial functions of dietary EPL. The purpose of this study was to elucidate the mechanism underlying the inhibition of neutral and acidic sterol absorption and hepatic enzyme activity-related fatty acid biosynthesis following EPL intake. EPL digest prepared using an in vitro digestion model had lower lipase activity and micellar lipid solubility and higher bile acid binding capacity than casein digest. Male Wistar rats were fed an AIN-93G diet containing 1% (wt/wt) EPL or L-lysine. After 4 weeks of feeding these diets, the marked decrease in serum and liver triacylglycerol contents by the EPL diet was partly attributed to increased fecal fatty acid excretion. The activities of hepatic acetyl-coenzyme A carboxylase and glucose-6-phosphate dehydrogenase, which are key enzymes of fatty acid biosynthesis, were enhanced in rats fed EPL diet. The increased fatty acid biosynthesis activity due to dietary EPL may be prevented by the enhancement of fecal fatty acid excretion. The hypocholesterolemic effect of EPL was mediated by increased fecal neutral and acidic sterol excretions due to the EPL digest suppressing micellar lipid solubility and high bile acid binding capacity. These results show that dietary EPL has beneficial effects that could help prevent lifestyle-related diseases such as hyperlipidemia and atherosclerosis.

Pentachlorophenol impact assessment of haematological parameters in olive flounder, Paralichthys olivaceus

  • Jee, Jung-Hoon;Min, Eun-Young;Park, Hee-Ju;Lee, Ok-Hyun;Keum, Yoo-Hwa;Kim, Su-Mi;Woo, Sung-Ho;Park, Soo-Il;Kang, Ju-Chan
    • Journal of fish pathology
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    • v.18 no.1
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    • pp.81-89
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    • 2005
  • The toxic effects of pentachlorophenol (PCP) on haematological parameters in olive flounder (Paralichthys olivaceus) after chronic exposure to dietary PCP (0.1, 0.5 and 1.0 mg/kg diet) for 2, 4 and 6 weeks were studied. A significant decrease in total RBC count, haemoglobin concentration and hematocrit value was noted in fish exposed to PCP compared to the non-exposed fish. The PCP treatment group showed significantly lower concentration of serum total protein and albumin, and significantly higher serum chloride, magnesium and total bilirubin levels compared with those in the control group. However, PCP had no major effects on serum glucose, total cholesterol, phosphate and calcium ions in flounder. These results demonstrated that PCP have induced adverse haematological impacts in the olive flounder, Paralichthys olivaceus. Because we found damages in blood-forming function and disruption in blood homeostasis due to chronic exposure to PCP, it is needed to develop further experimental studies for the risk assessment of this environmental pollutant.

Neuroprotective effects of Angelicae Acutilobae Radix water extract against ischemia·reperfusion-induced apoptosis in SK-N-SH neuronal cells (허혈·재관류 유도 신경세포사멸에 대한 일당귀 물추출물의 신경보호효과 연구)

  • Oh, Tae-Woo;Park, Ki-Ho;Lee, Mi-Young;Choi, Go-Ya;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.26 no.4
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    • pp.67-74
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    • 2011
  • Objectives : The purpose of the study is to determine the neuroprotective effects of the water extract of Angelicae Acutilobae Radix(AA) on ischemia reperfusion-induced apoptosis in SK-N-SH human brain neuronal cells. Methods: SK-N-SH cells were treated with different concentrations of AA water extract (0.1, 0.2, 0.5 and 1.0 mg/ml) for 2 hr and then stimulated with Dulbecco's phosphate-buffered saline containing CI-DPBS: 3mM sodium azide and 10 mM 2-deoxy-D-glucose for 45 min, reperfused with growth medium, and incubated for 24 h. Cell viability was determined by WST-1 assay, and ATP/ADP levels were measured by ADP/ATP ratio assay kit. The levels of caspase-3 protein were determined by Western blot and apoptotic body was observed by Hoechst 33258 staining. Results : AA extract significantly inhibited decreasing the cell viability in ischemia-induced SK-N-SH cells. AA also increased the ratio of ADP/ATP in ischemia-induced neuronal cells and decreased the expression levels of apoptotic protein, caspase-3 and apoptotic DNA damage. Conclusions : Our results suggest that AA extract has a neuroprotective property via suppressing the apoptosis and increasing the energy levels in neuronal cells, suggesting that AA extract may has a therapeutic potential in the treatment of ischemic brain injury.

Effect on Endurance Improving of Mixture of the Extracts of Watermelon and Octacosanol (Octacosanol과 박과식물 추출물을 주성분으로 하는 혼합물의 지구력 증진 효과)

  • Lim, Hyun-Woo;Kim, Sung-Kyu;Lee, Min-Won
    • YAKHAK HOEJI
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    • v.49 no.6
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    • pp.465-470
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    • 2005
  • Octacosanol is known to enhance endurance activities, control cholesterol in body and improve the function of cardiopulmonary. Citrulline, which is main compound of watermelon, is known to improve angiectasia through stimulating production of nitric oxide. To improve endurance activity swimming test on rats was carried out using four samples such as 1$\%$ octacosanol, citrulline, the extracts of barks of watermelon and products, mixture of 1$\%$ octacosanol and the extracts of barks of watermelon (6 : 4). Biochemical assays on the liver and serum of tested rats were also performed using commercial analysis kits. In result, it was shown that swimming time of III group increased by 26$\%$ and that of V group was increased by 22$\%$ at the swimming test. As a result of biological assays on the liver and serum of tested rats it was possible to confirm stability of toxicity When compared with creatine kinase of control group (549.11$\pm$39.15 U/l) citrulline (644.11 $\pm$50.67 U/l) and products group (646.00$\pm$46.99 U/l) were largely increased. When compared with inorganic phosphate of control group (12.01$\pm$0.75 mg/이), citrulline (13.03$\pm$0.94 mg/dl) and products group (12.90$\pm$0.55 mg/dl) showed similar results. Also, when compared with lactic acid and glucose of control group (152.91 $\pm$ 13.45, 103.00$\pm$ 8.69 mg/dl), citrulline (125.53$\pm$15.54, 83.75$\pm$7.29 mg/dl) and products group (135.26$\pm$11.50, 78.57$\pm$9.79 mg/dl) were largely decreased. As these test results, it was determined that 1$\%$ octacosanol and extracts of barks of watermelon had some effect of improving endurance activity. Futhermore, it was thought that it could be used as source of functional food.

Seasons affect the phosphorylation of pork sarcoplasmic proteins related to meat quality

  • Zeng, Xianming;Li, Xiao;Li, Chunbao
    • Animal Bioscience
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    • v.35 no.1
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    • pp.96-104
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    • 2022
  • Objective: Sarcoplasmic proteins include proteins that play critical roles in biological processes of living organisms. How seasons influence biological processes and meat quality of postmortem muscles through the regulation of protein phosphorylation remain to be investigated. In this study, the phosphorylation of sarcoplasmic proteins in pork longissimus muscle was investigated in four seasons. Methods: Sarcoplasmic proteins were extracted from 40 pork carcasses (10 for each season) and analyzed through ProQ Diamond staining for phosphorylation labeling and Sypro Ruby staining for total protein labeling. The pH of muscle, contents of glycogen and ATP were measured at 45 min, 3 h, and 9 h postmortem and the water (P2b, P21, and P22) was measured at 3 h and 9 h. Results: A total of 21 bands were detected. Band 8 (heat shock cognate 71 kDa protein; heat shock 70 kDa protein 1B) had higher phosphorylation level in summer than that in other seasons at 45 min postmortem. The phosphorylation levels of 3 Bands were significantly different between fast and normal pH decline groups (p<0.05). The phosphorylation levels of 4 bands showed negative associations with immobilized water (P21) and positive association with free water (P22). Conclusion: The phosphorylation levels of sarcoplasmic proteins involved in energy metabolism and heat stress response at early postmortem time differed depending on the seasons. These proteins include heat shock protein 70, pyruvate kinase, phosphoglucomutase-1, glucose-6-phosphate isomerase, and carbonic anhydrase 3. High temperatures in summer might result in the phosphorylation of those proteins, leading to pH decline and low water holding capacity.

Effects of prolonged photoperiod on growth performance, serum lipids and meat quality of Jinjiang cattle in winter

  • Yu, Yan;Qiu, Jingyun;Cao, Jincheng;Guo, Yingying;Bai, Hui;Wei, Shengjuan;Yan, Peishi
    • Animal Bioscience
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    • v.34 no.9
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    • pp.1569-1578
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    • 2021
  • Objective: This study was conducted to investigate the potential effects of prolonged photoperiod on the serum lipids, carcass traits, and meat quality of Jinjiang cattle during winter. Methods: Thirty-four Jinjiang bulls aged between 14 and 16 months were randomly assigned to two groups that were alternatively subjected to either natural daylight +4 h supplemental light (long photoperiod, LP) or natural daylight (natural photoperiod, NP) for 96 days. The potential effects on the levels of serum lipids, carcass traits, meat quality, and genes regulating lipid metabolism in the intramuscular fat (IMF) of the cattle were evaluated. Results: Jinjiang cattle kept under LP showed significant increase in both dry matter intake and backfat thickness. the serum glucose and the plasma leptin levels were significantly reduced, while that of melatonin and insulin were observed to be increased. The crude fat contents of biceps femoris muscle and longissimus dorsi muscle were higher in LP than in NP group. In longissimus dorsi muscle, the proportions of C17:0 and C18:0 were significantly higher but that of the C16:1 was found to be significantly lower in LP group. The relative mRNA expressions in IMF of longissimus dorsi muscle, the lipid synthesis genes (proliferator-activated receptor gamma, fatty acid-binding protein) and the fatty acid synthesis genes (acetyl-coa carboxylase, fatty acid synthetase, 1-acylglycerol-3-phosphate acyltransferase) were significantly up-regulated in LP group (p<0.05); whereas the hormone-sensitive lipase and stearoyl-CoA desaturase 1 were significantly down-regulated in LP than in NP group. Conclusion: Prolonged photoperiod significantly altered the growth performance, hormonal levels, gene expression and fat deposition in Jinjiang cattle. It suggested that the LP improved the fat deposition by regulating the levels of different hormones and genes related to lipid metabolism, thereby improving the fattening of Jinjiang cattle during winter.

Effects of Green Tea Extract on Intestinal Mucosal Esterification of $^{14}C$-Oleic Acid in Rats (녹차 추출물이 흰쥐 소장세포의 지방 에스테르화 과정에 미치는 영향)

  • Seo, Yun-Jung;Noh, Sang-K.
    • Food Science and Preservation
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    • v.15 no.3
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    • pp.450-455
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    • 2008
  • Previously, we have shown that green tea extract lowers the intestinal absorption of cholesterol, fat, and other fat-soluble compounds. We conducted this study to determine whether green tea extract affects the rate of $^{14}C$-oleic acid esterification into various lipids in the intestinal mucosa of rats. Male Sprague-Dawley ruts were had free access to a nutritionally adequate AIN-93G diet and deionized water. Initially, the rat's mucosal content of total lipids was measured following 1 mL olive oil administration with (green tea group) or without (control group) 100 mg green tea extract powder. At 1 h and 5 h, intestinal segments were extracted for total lipid analysis. Secondly, to measure mucosal esterification rates of lipids, an abdominal incision was made along the midline, and a 10-cm long jejunal segment of the small intestine was ligated in situ. Then, micellar solutions with or without green tea extract were injected into the ligated jejunal segments and incubated for 10 mill. The micellar solution contained $200.0\;{\mu}$ Ci $^{14}C$-oleic acid, $200.1\;{\mu}mol$ unlabelled oleic acid, $66.7\;{\mu}mol$ 2-monooleoylglycerol, $66.7\;{\mu}mol$ palmitoyl-sn-glycero-3-phosphocholine, 2.2 mmol glucose, $50.0\;{\mu}mol$ albumin, and 16.5 mmol Na-taurocholate per L of phosphate buffered saline (pH, 6.3) with or without 8.87 g green tea extract powder. At 10 min, each rat was sacrificed by cervical dislocation under anesthesia and the segment was removed for lipid analysis. Significant differences were observed in mucosal triglyceride content at 1 h and 5 h in ruts given green tea extract. Significant differences in the rate of $^{14}C$-oleic acid esterification into triglycerides and phospholipids fractions were observed between control and green tea groups. However, There were no significant differences in other lipid fractions. These results indicate that the lowered esterification rates of $^{14}C$-oleic acid into triglycerides and phospholipids fractions is attributable to presence of green tea extract. This may be associated with an inhibitory effect of green tea catechin on the mucosal processes of lipids, leading to the inhibition of intestinal absorption of lipids.