• Title/Summary/Keyword: Amino acid ester

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Ionomycin Recovers Taurine Transporter Activity in Cyclosporin A Treated macrophages

  • Kim, Ha-Won;Lee, Eun-Jin;Kim, Won-Bae;Hyun, Jin -Won;Kim, Byung-Kak
    • Preventive Nutrition and Food Science
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    • v.4 no.4
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    • pp.265-269
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    • 1999
  • Taurine is a major $\beta$-amino acid in various tissues. Taurine transporter (TAUT) is responsible for the transportation of taurine in the cell. The transporter is affected by various stimuli to maintain its cell volume. Macrophage cell volume varies in its activated states. In our experiment, it was found that the murine macrophage cell line, RAW264.7, expressed TAUT protein in its membrane. Its transportation activities could be blocked by a $\beta$-amino acid such as $\beta$-alanine, but not by $\alpha$-amino acids in this cell line. When assessed in RAW264.7 under the influence of immunosuppressive reagents, the activity of the TAUT was decreased by the treatment of rapamycin (RM) or cyclosporin A (CsA). However when ionomycin (IM) was added to this system, TAUT activity was recovered only in CsA-treated cells in a concentration-dependent manner. In order to inhibit the voltage gated {TEX}$Ca^{+2}${/TEX} channel, calmidazolium was added to the RAW264.7 cell line. Treatment of the cell with calmidazolium completely blocked TAUT. Furthermore, addition of IM to this system recovered the activity of TAUT again. When we added phorbol myristate acetate (PMA) to the cell line, secretion of nitric oxide (NO) was increased 4-fold and the TAUT activity was decreased 5-fold. However, the addition of N-nitro L-arginine methyl ester (L-NAME), an inducible NO synthase (iNOS) inhibitor, to the PMA-treated cells, resulted in the recovery of TAUT activity. These results showed that the activity of TAUT was sensitive to the intracellular concentrations of both {TEX}$Ca^{+2}${/TEX} and NO.

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Food Components of Wild and Cultured Fresh Water Fishes (천연 및 양식산 담수어의 식품성분)

  • KIM Kyung-Sam;LEE Eung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.19 no.3
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    • pp.195-211
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    • 1986
  • The object of this study is to obtain fundamental data on cultured fishes produced in Korea to improve their food components. For this purpose, the food components of cultured fresh water fishes such as eel, Anguilla japonica, snakehead, Channa argus, and common carp, Cyprinus carpio, were investigated and compared with those of the wild ones. The results obtained are summarized as follows: 1. Common characteristics in the proximate composition were that wild fish was higher in crude protein content and lower in crude lipid content than those of cultured one. 2. Among the 9 kinds of minerals analyzed in all the samples, sodium, potassium, calcium and magnesium contents were absolutely predominant being more than $99.52\%$. These four elements in feedstuff also occupied $99.68{\sim}99.92%$ of total minerals. 3. The neutral lipids of wild and cultured eel, snakehead and common carp occupied $55.7{\sim}95.8%$ of lipid fractions, while the content of the phospholipids in snakehead was particularly higher than those of others. 4. The neutral lipids of wild and cultured eel, snakehead and common carp mainly consisted of triglycerides ($85{\sim}95%$), and a little quantity of diglycerides, monoglycerides, free sterol ester and hydrocarbon were also identified in the neutral lipid. 5. The phospolipids of eel and common carp were mainly occupied by phosphatidyl choline ($71.3{\sim}83.9%$), followed by phosphatidyl ethanolamine ($12.1{\sim}23.5%$) and phosphatidyl serine ($7.5{\sim}13.8%$). The phospholipids of snakhead consisted of phosphatidyl choline ($50.7{\sim}64.5%$), phosphatidyl ethanolamine ($28.0{\sim}35.5%$) and phosphatidyl serine ($7.5{\sim}13.8%$). Generally, phosphatidyl choline content was higher in wild fish than in cultured one, while phosphatidyl ethanolamine and phosphatidyl serine contents were higher in cultured one. 6. The major fatty acids in total lipid of wild eel, snakehead and common carp were $C_{16:0}\;and\;C_{20:5}$, while those in cultured ones were $C_{18:1},\;C_{18:2}\;and\;C_{22:6}$. The fatty acid composition of neutral lipids showed similar tendency to that of total lipid, and the main fatty acids in phospholipids of cultured fishes were $C_{18:1}\;and\;C_{18:2}$. In glycolipids, $C_{20:5}\;and\;C_{22:6}$ were higher in wild fishes, while $C_{18:2}$ were higher in cultured ones. 7. Total amino acids contents of wild and cultured eel were nearly the same, being $16.65\%$ ana $15.99\%$ respectively. The major amino acids of wild and cultured fish were glutamic acid, leucine, aspartic acid and lysine in order. In snakehead, the contents of aspartic acid and proline in cultured fish were higher than those in wild one, while the contents of glutamic acid, alanine, glycine were higher in the wild one. Total amino acid content of cultured common carp was $21.7\%$ compared with $17.08\%$ in wild one. The contents of glutamic acid, aspartic acid, glycine, proline and alanine occupied higher quantities in cultured common carp compared with those in wild one while the other amino acids revealed no significant difference. 8. Aspartic acid in free amino acids of cultured eel held $1.0\%$ of total free amino acids, while that in wild eel held $2.9\%$. Histidine, arginine and tyrosine content of cultured fish were two times higher than those of wild one. But free amino acid composition of samples seemed to be no marked differences according to cultured places. The contents of arginine, aspartic acid, glutamic acid, methionine and phenylalanine of snakehead ware higher in wild one than in cultured one, while the contents of lysine, histidine, glycine, and alanine ware higher in cultured one. In free amino acids content of wild common carp, histidine, glycine and lysine occupied $76.9\%$ of total free amino acids. Lysine, histidine, aspartic acid, alanine, valine and leucine were higher in wild one compared with those of cultured one, while glycine and tyrosine contents were higher in cultured fish.

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Peptide Synthesis with Polymer Bound Active Ester III. The Effect of Spacer Arm in Peptide Synthesis with 1-Phenyl-3-methyl-4-oximino pyrazole Resin (고분자에 결합된 활성 에스테르에 의한 펩티드합성 III. 1-페닐-3-메틸-4-옥시미노피라졸레진을 이용한 펩티드 합성에서 Spacer Arm의 효과)

  • Wang, Young;Lee, Yoon-Sik
    • Applied Chemistry for Engineering
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    • v.4 no.1
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    • pp.132-143
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    • 1993
  • Aminomethyl polystyrene resins were prepared either from chloromethyl-resin(Merrifield resin) or from direct amidoalkylation of polystyrene resin. Two kinds of aminomethlyl resin were lengthened with spacer arms via sequential coupling of five ${\varepsilon}$-aminocaproic acids(ACA) respectively. In case of the resin prepared from the Merrifield resin, the amounts of free amino group of the resin were reduced by 25~30% after each coupling of ACA. But the one from direct amidoalkylation showed 3~5% loss after each coupling of ACA. 4-Nitroso-5-aminopyrazole resin was made by reacting ACA spacer arm resin, which was made from direct amidoalkylated resin, with 5-phenyl-7-methylpyrazole [4,3-c][1,2,4]oxadiazin-3-one. Several polymeric active esters of N-blocked amino acids were prepared from the 4-nitroso-5-aminopyrazole bound resins. In anchoring step of the amino acid derivatives on the resin, no substantial effect of bulkiness was found. 4-Nitroso-5-aminopyrazole bound active ester resins were found to be very reactive in N-acylation, The resulting peptides were obtained with 90~95% yield and characterized by NMR and other physical methods.

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Studies on Lao-Chao Culture Filtrate for a Flavoring Agent in a Yogurt-Like Product

  • Liu, Yi-Chung;Chen, Ming-Ju;Lin, Chin-Wen
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.4
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    • pp.602-609
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    • 2002
  • Lao-chao is a traditional Chinese fermented rice product with a sweet and fruity flavor, containing high levels of glucose, a little alcohol and milk-clotting characteristics. In order to optimize commercial production of lao-chao, Rhizopus javanicus and Saccharomyces cerevisiae were selected as the mold and yeast starter, respectively. A commercial mixed starter (chiu-yao) was used as control. Fermentation of the experimental combination revealed a sharp drop in pH (to 4.5) on the fourth day, remaining constant thereafter. Content of reducing sugars gradually decreased throughout the entire fermentation period. Of the free amino acids, higher quantities of alanine, leucine, proline, glutamic acid, glutamine and $NH_3$ were noted. For sugars, glucose revealed the highest concentration, while organic acid levels, including those for oxalic, lactic, citric and pyroglutamic acid, increased throughout the fermentation period. Twenty-one compounds were identified by gas chromatography from aroma concentrates of the lao-chao culture filtrate, prepared using the headspace method. For the flavor components, higher quantities of ethanol, fusel oil and ester were determined in both culture filtrates. In regard to the evaluation of yogurt-like product, there were significant differences in alcoholic smell, texture and curd firmness.

Development of Soluble Epoxide Hydrolase Inhibitor Screening Methods for Discovery of Drug Candidate in Cardiovascular Diseases (심혈관계 질환 치료제 후보물질 발굴을 위한 Soluble Epoxide Hydrolase 억제평가 방법 개발)

  • Lee, Gwan-Ho;Kim, Bong-Hee;Kim, Sang-Kyum
    • YAKHAK HOEJI
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    • v.56 no.1
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    • pp.42-47
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    • 2012
  • Soluble epoxide hydrolase (sEH) is a metabolic regulator of epoxyeicosatrienoic acids (EETs). EETs have many beneficial effects, vasodilation, anti-diabetes, anti-inflammation, cardiovascular protection, renal protection. Therefore, selective sEH inhibitors have a potential for treating these diseases. In the present study, screening methods for sEH inhibitors using PHOME ((3-phenyl-oxiranyl)-acetic acid cyano-(6-methoxynaphthalen-2-yl)-methyl ester) and 14-15-EET as substrates were established. To determine selectivity, microsomal epoxide hydrolase (mEH) inhibition assay was also developed using styrene oxide as a substrate of microsomal epoxide hydrolase. Our results obtained from 12-[[(tricyclo[3.3.1.13,7]dec-1-ylamino)carbonyl]amino]-dodecanoic acid (AUDA) used as a positive sEH inhibitor and valpromide used as a positive mEH inhibitor showed that these methods are useful for discovery of drug candidates.

Quality characteristics of fermented soybean products produced by lactic acid bacteria isolated from traditional soybean paste (전통 장류 유래 유산균을 이용한 콩 발효물의 품질특성)

  • Lee, Sun Young;Seo, Bo Young;Eom, Jeong Seon;Choi, Hye Sun
    • Food Science and Preservation
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    • v.24 no.2
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    • pp.187-195
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    • 2017
  • This study evaluated quality characteristics of soybean fermented by selected lactic acid bacteria, which were the enzyme strains with high antimicrobial activities isolated from traditionally prepared soybean paste. We determined total aerobic and lactic acid bacteria counts, protease and amylase activities, reducing sugar and amino-type nitrogen contents, and the amounts of amino acids, organic acids, and aroma-compounds. The total aerobic bacteria counts in soybean fermented with strain I13 ($7.75{\times}10^9CFU/mL$) were the highest among all the strains analyzed. Lactic acid bacteria numbers were $2.85{\times}10^9$ to $4.35{\times}10^9CFU/mL$ in soybean fermented with isolates. Amylase and protease activities of the JSB22 sample were the highest among all sample. Reducing sugar and amino-type nitrogen contents of soybean fermented with JSB22 (1.23%, 94.52 mg%) were highest. Total amino acid content of the samples was 15.88-17.62%, and glutamic acid, aspartic acid, leucine, lysine, and arginine were the major amino acids. Lactic acid (0.82-3.65 g/100 g), oxalic acid (22.74-63.57 mg/100 g), and fumaric acid (2.88-6.33 mg/100 g) were predominant organic acids. A total of 39 volatile aroma-compounds were identified, including 2 esters, 5 ketones, 7 alcohols, 14 hydrocarbons, 2 heterocyclic compounds, 4 acids, and 5 miscellaneous compounds. These results represent useful information for the development of a starter (single or complex) and will be used for production of functional fermented soybean foods.

Potential Utilization of Concentrated Oyster Cooker Effluent for Seafood Flavoring Agent (수산식품 조미제 개발을 위한 굴 자숙 농축액의 이용)

  • KIM Jeong Han
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.1
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    • pp.79-85
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    • 2000
  • Proximate composition of concentrated oyster cooker effluent (COCE) was $70{\%}$ moisture, $8.55{\%}$ total nitrogen, and $18.6{\%}$ ash. Optimum conditions for enzymatic hydrolysis of COCE ($200 ml$) weve $pH 7.0{\~}7.5\;and 50{\~}60{\circ}C$ for 8 h reaction time with 0.128 U of Aspergillus oryzae PE protease(AOP), Hydrolysis of COCE led to an increase in free amino acids to 1.41 fold, with taurine comprising about $17.62{\%}$ of the total free amino acid. Fifty volatile flavor components were identified in COCE and 63 in enzyme treated COCE (HCOCE). Volatile flayer compounds we increased significantly in HCOCE when compared with untreated COCE. Results of this study might help to alleviate the current wastewater disposal problem caused by oyster cooker effluent.

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A Cloning of Novel Esterase from a Metagenomic Library

  • Yoon, Sang-Young;Kim, Seung-Bum;Ryu, Yeon-Woo
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.243-246
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    • 2005
  • A novel esterase showing high enantioselectivity to (S)-ketoprofen ethyl ester was selected from fosmid environmental DNA library which is provided by Microbial Genomic & Applications Center. As a result of Blast search, the gene wasn't registerated in Gene Bank yet. And as we know, conserved domain region of esterase , G-X-S-X-G, wasn't discovered.$^{4)}$ And it is similar to Beta-lactamase. The DNA sequence of cloned esterase include an open reading frame consisting of 1170 bp, designated as EST-Y29, encoding a protein of 389 amino acids with a molecular mass of about 42.8 kDa. And amino acid sequence analysis revealed only a few identity (28%) to tile known esterases/lipases in the databases containing the conserved sequence motifs of esterases/lipases. when being comparison to other esterase revealed , this enzyme seems to be classified as a new member of esterase family. EST-Y29 was functionally overexpressed in a soluble form in E. coli with maximum conversion yield of (S)-ketoprofen at $65^{\circ}C$. This study demonstrates that functional screening combined with the sequential uses of restriction enzymes to exclude already known enzymes is a useful approach for isolating novel enzyme from a metagenome.

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Synthesis of Insulin A (1-21) Chain and Their Assembly on a Polymer-Bound ${\alpha}$-Methylphenacylester Linkage (2-브로모프로피오닐화된 수지를 이용한 인슐린 A (1-21) 사슬의 합성)

  • Soon Uoong Koock;Nam-Joo Hong
    • Journal of the Korean Chemical Society
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    • v.31 no.5
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    • pp.457-463
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    • 1987
  • The total synthesis of insulin A chain (1-21) with properly protected sulfhdryl groups of three cysteins for the correct intra and inter disulfide bond formation has been accomplished on 2-bromopropionylated 2% DVB-styreneresin support employing manually operated rotary vessel. The sulfhydryl groups of cysteins were protected with acetamidomethyl, benzyl, and benzhydryl respectively. Glutamine and asparagine were attached to the peptide chain by active ester coupling, all other amino acids were coupled with DCC/HOBT. The synthesized peptide was purified by DEAE Sephadex A-25 and gel filtration Sephadex LH-20. The final product was found to be homogeneous by HPLC, electrophoresis, and amino acid analysis. The overall yield of the pure isolated peptide was 6%.

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Investigation into the Efficacy of Val-SN-38, a Valine-Ester Prodrug of the Anti-Cancer Agent SN-38

  • Kwak, Eun-Young;Choi, Min-Koo;Yang, Su-Geun;Shim, Chang-Koo;Shim, Won-Sik
    • Biomolecules & Therapeutics
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    • v.20 no.3
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    • pp.326-331
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    • 2012
  • We recently reported that Val-SN-38, a novel valine ester prodrug of SN-38, had greatly improved the intracellular accumulation of SN-38 in MCF-7 cell line, probably through enhanced uptake via amino acid transporters. In the present study, the efficacy of Val-SN-38 was further investigated both in vitro and in vivo. It was found that the in vitro cytotoxic effect of Val-SN-38 was similar to that of SN-38. Moreover, Val-SN-38 exhibited an equal potency to that of SN-38 in survival experiments in vivo. Because these results seemed to be contrary to the previous finding, further investigation was performed to find out the underlying cause of the contradiction. As only the lactone form is known to have cytotoxic activity, the proportion of lactone in Val-SN-38 and SN-38 was determined, but no differences were found. However, it turned out that Val-SN-38 had poor stability compared with SN-38, which resulted in a decrease in beneficial efficacy for Val-SN-38. Overall, the present study showed that a valine-added prodrug approach could be advantageous provided that the stability of the compound can be ensured. We believe this is a noteworthy study that unravels the discrepancy between intracellular accumulation and efficacy of valine-added prodrug.