• KSBB Journal
• /
• v.26 no.5
• /
• pp.386-392
• /
• 2011

Human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig), an immunosuppressive agent, was expressed in transgenic rice cells using RAmy3D promoter and RAmy1A signal peptide for the inducible production and secretion into culture media by sugar depletion. In this study, sodium butyrate was used as a small molecular enhancer (SME) to enhance the production of hCTLA4Ig in transgenic rice cell suspension cultures. When 1 mM sodium butyrate was added in sugar-free media, relative viability was not reduced, while the productivity was improved 1.3-fold. In addition, by supplementing 87 mM sodium pyruvate as an alternative energy source during the production phase, death rate of the cells was decreased. When sodium pyruvate was not added, most cells became dead at day 6. However, by adding sodium pyruvate, 18% of viability can be maintained until day 10 and the production of hCTLA4Ig was enhanced 1.4-fold. When the combination of sodium pyruvate and sodium butyrate at optimum concentrations was added, the highest viability and hCTLA4Ig production could be obtained. The highest level of hCTLA4Ig reached up to 35 mg/L at day 10.

• Proceedings of the KSAR Conference
• /
• 2003.06a
• /
• pp.64-64
• /
• 2003

본 연구는 돼지의 미성숙란을 체외에서 성숙, 수정시킨 후 생산된 체외수정란을 NCSU23 체외배양액에 sodium pyruvate, hemicalcium lactate 및 glucose를 첨가해 돼지 체외수정란의 체외발육에 미치는 영향을 검토하였다. NCSU23 체외배양액에 Glucose 0 mM, 2.78 mM, 5.56 mM 및 11.12 mM을 처리하여 체외배양한 결과 분할율은 glucose처리구가 무처리구에 비해 통계적으로 유의하게 높은 성적을 나타냈으며(P<0.05), 상실배까지 체외발육률 또한 glucose처리구가 무처리구에 비해 통계적으로 유의하게 높은 성적을 나타냈다 (P<0.05). 한편 배반포까지의 체외발육율은 5.56 mM 처리구가 다른 처리구에 비해 높은 경향을 나타냈지만 통계적 유의성은 인정되지 않았다. Glucose와 0.4mM sodium pyruvate, 2mM hemicalcium lactate 공동배양 처리구와 glucose를 첨가하지 않은 처리구의 분할율은 처리구간 커다란 차이를 나타내지 않았으며, 상실배기 이상 발육된 체외 발육율도 처리구간 커다란 차이를 나타내지 않았다. 또한, 각 처리구간 분할율은 hemicalcium lactate와 glucose공동배양 처리구가 가장 좋은 성적을 나타냈지만 통계적 유의성은 인정되지 않았으며, 상실배기 이상 발육된 체외 발육율은 hemicalcium lactate만 첨가한 처리구가 가장 좋은 성적을 나타냈지만 통계적 유의성은 인정되지 않았다. Sodium pyruvate를 체외수정 후 8시간, 48시간, 96시간, 120시간에 제거하였을 때 분할율은 sodium pyruvate를 제거하는 시기가 늦어질수록 분할율이 향상되는 경향을 나타냈지만 통계적 유의성은 인정되지 않았다. 또한, 상실배기 이상 발육된 체외 발육율은 제거하는 시기가 늦어질수록 체외 발육율이 향상되는 경향을 나타냈지만 통계적 유의성은 인정되지 않았다. 본 연구의 결과 돼지 체외수정란의 체외배양시 glucose의 첨가는 체외발육을 증진시키는 결과가 있었으며, glucose와 sodium pyruvate, hemicalcium lactate 공동배양은 체외배양에 별다른 영향을 미치지 못하는 것으로 나타났다.

• Reproductive and Developmental Biology
• /
• v.30 no.1
• /
• pp.7-11
• /
• 2006

This study was conducted to examine the effects of hormone and sodium pyruvate on in vitro maturation of canine oocytes. Canine oocytes were collected from the ovaries of dogs and cultured in NCSU-37 medium with hormones and sodium pyruvate for 72 hr. Oocytes matured to the metaphase II (MII) stage were observed only from estradiol $17{\beta}\;(E_2)$, and the presence of gonadotropin did not improve the nuclear maturation. No oocytes were developed to the MII stage when $E_2$ was added to medium during the first 6 and 24 hrs of culture period. The presence of $E_2$ during the whole culture period enhanced the nuclear maturation to the MII stage (6.0%, P<0.05). High concentration of sodium pyruvate (2.5 mM) slightly enhanced the nuclear maturation to the metapahse I (HMI) stage, but not the MII stage. the result of the present study shows that the presence of $ E_2$ during the whole culture period of 72 hr enhances the maturation of canine oocytes to the M stage, but sodium pyruvate does not affect the nuclear maturation of the canine oocytes.

• The Korean Journal of Pharmacology
• /
• v.10 no.1 s.15
• /
• pp.21-39
• /
• 1974

Certain metabolic aspects of halothane's cardiac depressant action on the contractility of the myocardium were elucidated from a sudy of the effect of pyruvate on halothane-depressed rat atria. Approximately 6 mg% halothane was required to maintain a 50% depression of the contractility of rat atria suspended in a modified Krebs-Ringer bicarbonate glucose medium, pH 7.4, $30^{\circ}C$ for a 2 hr. period. Pyruvate was found to restore partially the contractility of halothane-depressed atria. The maximally effective concentration of pyruvate was 2.5 mM. There was minimal pyruvate effect on the force of contraction of control atria. The effect of pyruvate on halothane-depressed atria was shown to be due to the pyruvate and not the sodium ion of the sodium pyruvate. Pyruvate was found to produce no increase in the contractility of atria depressed by hypertonic medium, but caused a further depression. Selected aspects regarding the action of halothane on glucose metabolism in myocardial cells are discussed. The results are consistent with the hypothesis that at least a part of the negative inotropic action of halothane is due to an inhibition of glucose uptake or utilization in the glycolytic pathway.

• The Korean Journal of Pharmacology
• /
• v.28 no.1
• /
• pp.41-48
• /
• 1992

A concentration of 0.01 mM bupivacaine was necessary to maintain approximately 50% depression of contractility of rat atria suspended in a modified Krebs-Ringer bicarbonate glucose medium, pH 7.4 at $30^{\circ}C$. Sodium pyruvate, sodium acetate, and fructose partially restored the contractility of the bupivacaine-depressed atria. However, 20 mM glucose had no effect on the bupivacaine-depressed atria, although this concentration of glucose markedly increased the contractility of normal atria not to be exposed to bupivacaine. Contractility of normal atria was not significantly influenced by sodium pyruvate, sodium acetate, and fructose. The results suggested that at least part of the negative inotropic action of bupivacaine is the result of inhibition of glucose uptake or utilization in the glycolytic pathway, and further pinpoint the blockade as an early step in the glycolytic sequence prior to the phosphofructokinase step.

• Applied Biological Chemistry
• /
• v.35 no.1
• /
• pp.6-9
• /
• 1992

The browing, pyruvate content of sliced garlic during the drying in terms of drying temperature and sulfiting treatment were investigated. Sulfiting reduced the browning of garlic slices and prevented the reduction of pyruvate content against heat during the drying. Contents of pyruvate were decreased as the increase of drying temperature. Sulfiting treatment killed the microorganisms of sliced garlic products during the drying.

• Archives of Pharmacal Research
• /
• v.19 no.6
• /
• pp.502-506
• /
• 1996

Protein X is one of the subunits of pyruvate dehydrogenase complex. The biological role of this protein has not been fully elucidated, mainly because of the difficulty in its dissociation from the tightly bound dihydrolipoamide acetyltransferase-protein X subcomplex. We have found that the detachment of protein X from acetyltransferase subunit can be easily accomplished by the cycles of freezing and thawing proces. Several lines of evidence including sodium dodecyl sulfate-polyacrylamide gel electrophoresis, N-terminal amino acid sequence analysis and acetylation with $[2^{14}C]$ pyruvate confirmed that the purified protein is protein X. The purified intact form of protein X was acetylated by $[2^{14}C]$ pyruvate in the presence of py-ruvate dehydrogenase subunit.The acetylation efficiency of this protein was lower than that of acetyltransferase and was not affected by the presence of acetyltransferase.

• BMB Reports
• /
• v.28 no.2
• /
• pp.118-123
• /
• 1995

Biodegradative threonine dehydratase was purified to homogeneity from Serratia marcescens ATCC 25419 by streptomycin sulfate treatment, Sephadex G-200 gel filtration chromatography followed by AMP-Sepharose 4B affinity chromatography. The molecular weight of the purified enzyme was 118,000 by fast protein liquid chromatography using superose 6-HR. The enzyme was determined to be a homotetrameric protein with subunit molecular weights of 30,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme was inhibited by ${\alpha}-Keto$ acids and their derivatives such as ${\alpha}-ketobutyrate$, pyruvate, glyoxlyate, and phosphoenol pyruvate, but not by ${\alpha}-aminobutyrate$ and ${\alpha}-hydroxybutyrate$. The inhibition of the enzyme by pyruvate and glyoxylate was observed in the presence of AMP. The inhibitory effect of glyoxylate was decreased at high enzyme concentration, whereas the inhibition by pyruvate was independent of the enzyme concentration. The kinetics of inhibition of the enzyme by pyruvate and glyoxylate revealed a noncompetitive and mixed-type inhibition by the two inhibitors with respect to L-threonine and AMP, respectively.

• Journal of Life Science
• /
• v.14 no.5
• /
• pp.868-873
• /
• 2004

The efficient fermentative production of L-threonine fermentation was achieved by using Escherichia coli MT201, transformed a plasmid carrying pyruvate carboxylase gene. It is an attempt to supply oxaloacetate to the L-threonine biosynthetic pathway. In order to improve the L-threonine productivity of E. coli MT201, a plasmid pPYC which is an expression vector of the pyruvate carboxylase gene of Coryne-bacterium glutamicum, was introduced. When E. coli MT/pPYC was incubated with medium containing only glucose as a carbon source, both the cell growth and L-threonine production were reduced, compared to the results from fermentation of E. coli MT201. In order to circumvent this effect, we attempted the addition of a mixed carbon source, composed of glucose and sodium citrate at a ratio of 1.5:3.5. It was shown that L-threonine production and cell growth (OD660) with E. coli MT/pPYC reached up to 75.7 g/l and 48, respectively, at incubation for 75 hr under fed-batch fermentation conditions. It is assumed that overproduction of L-threonine by anaplerotic pathway leads unbalance of TCA cycle and sodium citrate might playa role to recover normal TCA cycle.

• Microbiology and Biotechnology Letters
• /
• v.24 no.1
• /
• pp.44-49
• /
• 1996

By using the ${\beta}$-tyrosinase of Citrobacter freundii KCT2006, which was cloned and overexpressed in Escherichia coli, 3,4-dihydroxy phenyl-L-alanine (L-DOPA) was synthesized efficiently from pyrocatechol, sodium pyruvate, and ammonium acetate. Optimal temperature and pH for the reaction were determined to be about 18$^{\circ}C$ and 8.5, respectively. The effects of substrate concentrations were also examined at different concentrations of ammonium acetate, sodium pyruvate, and pyrocatechol. Ammoniumacetate and sodium pyruvate increased the reaction rate until the concentrations reached to 300mM and 50mM, respectively. Although pyrocatechol showed the optimal concentration at 20mM, it was controlled between 20mM and 50mM to avoid the depletion of substrate during the enzymatic synthesis. Meanwhile the synthetic rate was improved about 20% when ethanol was included in the reaction solution. Based on above results, a reaction medium for the productin of L-DOPA was prepared and incubated with 1 unit/ml of ${\beta}$-tyrosinase. Pyrocatechol and sodium pyruvate was added to the reaction solutin intermittently to avoid the substrate depletion during the enzymatic reaction. After 24 hour of reaction, 31.6g/l of L-DOPA was accumulated in the reaction solution as soluble and precipitated ones and the conversion yield was about 85.2%.