• Title/Summary/Keyword: L-glutamine

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Fortification of Amino Acids to Improve Hybridoma Cell Growth and Monoclonal Antibody Production in Perfusion Culture (Perfusion배양시 세포성장 및 항체생산 향상을 위한 아미노산의 보강)

  • 이수영;최병욱;오한규;윤정원;전복환;변태호;박송용
    • KSBB Journal
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    • v.14 no.2
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    • pp.188-191
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    • 1999
  • We have investigated the fortifying effect of amino acids on the cell growth and productivity during the perfusion culture of hybridoma vR8 cells in serum-free media. Through the quantitative analysis of amino acids and metabolites in perfusion culture, we found that many amino acids(glutamine, histidine, arginine, methionine, isoleucine, leucine, phenylalanine, tryptophane) were heavily consumed at cell density of $1.06{\times}10^7$cells/mL. Due to amino acid depletion, cells died suddenly. So we supplemented the media with those amino acids by 30-170%. As a result, were could increase maximum cell density by 270%, average specific productivity by 175%, and average volumetric productivity by 560% in this fortified media, GC-HY-S2.

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Iron Binding Peptides from Casein Hydrolysates Produced by Alcalase (Casein으로부터 Alcalase에 의해 생성된 철분결합 Peptide)

  • Choi, In-Wook;Kim, Kee-Sung;Lim, Sang-Dong;Lim, Sin-Won
    • Korean Journal of Food Science and Technology
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    • v.30 no.1
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    • pp.218-223
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    • 1998
  • Casein was hydrolyzed by alcalase to produce iron binding peptide (IBP). IBP was effectively separated from casein hydrolysates by immobilized $Fe^{3+}$ affinity chromatography and further purified by reverse phase chromatography. $25,\;50\;and\;100\;{\mu}g/mL$ of IBP solubilized $4.2,\;5.7\;and\;7.1\;{\mu}g$ of ferric at duodenum condition $(pH\;6,\;37^{\circ}C)$, respectively. According to the result of MALDI analysis, molecular weight of IBP was determined to 2,175 dalton. IBP was mainly composed of proline (24.5 mol%), lysine (15.7 mol%), and glutamine or glutamic acid (14.9 mol%) and its N-terminal sequence was Met-Ala-Pro-Lys-His. According to the information obtained from molecular weight, amino acids composition and N-terminal sequence of IBP, it was evident that IBP was from f102-119 of ${\beta}-casein$.

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Purification and Properties of $\gamma$-Glutamyl Transpeptidase from Bacillus sp. KUN-17

  • Hwang, Se-Young;Ryang, Jun-Hwan;Lim, Wang-Jin;Yoo, Ick-Dong;Kunio Oishi
    • Journal of Microbiology and Biotechnology
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    • v.6 no.4
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    • pp.238-244
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    • 1996
  • $\gamma$-Glutamyl transpeptidase ($\gamma$-GTP; EC 2.3.2.2) present in the culture filtrate of Bacillus sp. KUN-17 was purified 400-fold through a consecutive procedure including organic precipitation and column chromatography. The enzyme has an estimated molecular weight of 70, 000 and consists of hetero-subunits with molecular weights of 42, 000 and 22, 000. In vitro optimal conditions for those transfer and hydrolysis reactions appeared to be pH 7.0 at $50^{\circ}C$ and pH 8.4 at $40^{\circ}C$, respectively. The denatured enzyme recovered most of its $\gamma$-GTP activity by removing detergents such as sodium dodecyl sulfate (SDS) or urea with dialysis. The enzyme showed higher affinities against a number of amino acids as $\gamma$-glutamyl acceptors than glycylglycine in the following order: L-valine, L-methionine, L-glutamic acid or L-as-paragine, L-alanine. Also, it was shown that L-glutamine was the most suitable $\gamma$-glutamyl donor for the transfer reaction among those tested. Amino acids generally inhibited the enzyme activity for the transfer reaction, but not for the hydrolysis reaction.

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N-Acyl Amino Acid Surfactant(15) Synthesis and Properties of Sodium N-(2-Dodecyl Succinoyl) l-Glutamate (N-아실아미노산계 계면활성제 (제15보) Sodium N-(2-Dodecyl Succinoyl) l-Glutamate의 합성 및 계면성)

  • Kwack, Kwang-Soo;Yoon, Young-Kyoon;Jeong, Noh-Hee;Kim, Duck-Gwon;Nam, Ki-Dae
    • Journal of the Korean Applied Science and Technology
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    • v.18 no.1
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    • pp.55-59
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    • 2001
  • These N-acyl amino acid surfactants is normally produced by reaction of acid anhydride with sodium ${\ell}-glutamate$ hydrolysates under Schotten-Baumann condition i.e., in alkaline aqueous medium. To avoid using fatty acid chlorides, acylations were also carried out with the fatty acids themselves or with their methyl esters, but unfortunately these methods cannot be used in practice, dodecenyl succinic anhydride, was to be studied for their suitability as acylating agents the production if acylated glutamine hydrolysates. The surface activities including surface tension forming power, forming stability and emulsifying power were measured. The experimental results revealed that the products have a good emulsifying power. Thus, there derivatives will be expected to be used an emulsifying agent for O/W type cosmetic emulsion.

Partial Desiccation of Embryogenic Calli Improves Plant Regeneration in Sugarcane (Saccharum Spp.)

  • Desai Neetin Shivajirao;Suprasanna Penna;Bapat Viswas Ananat
    • Journal of Plant Biotechnology
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    • v.6 no.4
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    • pp.229-233
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    • 2004
  • Partial desiccation of embryogenic calli cultures or somatic embryos leads to different physiological changes and maturation of somatic embryos, leading to improved plant regeneration. Embryogenic calli was induced from immature inflorescence segments and young leaf rolls of sugarcane (Saccharum officinarum hybrids CoC-671) on Murashige and Skoog's basal medium enriched with different concentrations of 2,4-D ($1-4\;\cal{mg/l}$), L-glutamine ($100\cal{mg/l}$), malt extract ($100\cal{mg/l}$), casein hydrolysate ($1000\;\cal{mg/l}$) and coconut milk ($5\%$) and solidified with $0.2\%$ gel rite. The embryogenic calli were subjected to desiccation for 1-8 h. Desiccation of the calli for 6-7 h resulted in enhancement of plant regeneration frequency ($83-96\%$) as compared to control ($12\%$). Plantlets exhibited vigorous growth to maturity in the greenhouse. Partial desiccation of embryogenic calli offers as a simple method for improving plant regeneration frequency in sugarcane.

Bioavailability and functions of L-glutamic acid (글루탐산의 생체이용률과 기능성)

  • Lee, Kwang-Won;Chun, Su-Hyun;Kim, HeeEun
    • Food Science and Industry
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    • v.50 no.3
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    • pp.93-104
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    • 2017
  • monosodium L-glutamate (MSG), composed of 88% L-glutamic acid (GLU) and 12% Na, is considered as Generally Recognized as Safe. GLU is accounting for 15% of the total amino acid content in our body. Daily GLU intake is large 3.5-10.6 g from food and 1.3 g from food additive. The daily formed GLU in the body, via degradation of the total protein, is 48-50 g (70 kg of male) and the maintained GLU in the body is total bound GLU (1.4-2.0 kg) and free GLU (10 g). About 88% GLU are consumed at digestive tract, and only 12% GLU enter the blood stream. GLU is generally known to precursors of N-acetyl glutamate and glutamine, substrates for protein synthesis, a neurotransmitter (GABA), and the active site of the enzyme. In addition to protein synthesis, GLU has these key functions within the body, thus this amino acid is critical for healthy body maintenance and function.

Interactions between Oxidative Pentose Phosphate Pathway and Enzymes of Nitrate Assimilation "Nitrate Reductase, Nitrite Reductase, Glutamine Synthetase$_1$" and Ammonium Reassimilation "Glutamine Synthetase$_2$" as affected by $No_3$-Concentration ($No_3$-수준이 Oxidative Pentose Phosphate Pathway와 질산동화작용 효소"Nitrate Reductase, Nitrite Reductase, Glutamine Synthetase$_1$" 및 암모늄재동화작용 주요효소"Glutamine Synthetase$_2$"활성도의 상호관계에 미치는 영향)

  • Sohn, Sang-Mok;Michael James Emes
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.37 no.5
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    • pp.468-475
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    • 1992
  • In order to understand more clearly the integration between N-assmilation and C-metabolism in relation to N fertilization, a pot experiment with 5 different level of N fertilization(0, 5, 10, 25, 50 mM NO$_3$$_{[-10]}$ ) was conducted in Manchester, U.K. The peas (Pisum sativum L., cv. Early Onward) were sown in vermiculate (5 cm depth) and cultivated for 6 days under temperature controlled dark room conditions ($25^{\circ}C$). The plants received frequent irrigation with a nutrient solution: it was fertilized every 2 days, 3 times a day at 10h, 13h, 16h respectively. Elevated NO$_3$$^{[-10]}$ concentration, the activity levels of NR, NiR, total GS(crude extract), GS$_2$(plastid) in both root and shoot were increased and reached the peak in 5~25 mM, except NiR specific activity which increased its activity continually until 50 mM NO$_3$$^{[-10]}$ treatment. Total activities of GS (crude extract) in both root and shoot became higher than those of GS$_2$(Plastid), and the activity ratios of total GS in the crude extract and GS$_2$ in the plastids were 3.0 to 4.3 in root, but 3.2 to 10.6 in shoot. It was concluded that the reductants and A TP from OPPP itself should be enough to achieve the high rate of NR, NiR, GS$_1$, GS$_2$ in plant root and shoot for reduction or assimilation of nitrogen, but these enzyme activities might be inhibited by an excess of NO$_3$$^{[-10]}$ influx over the reduction capacity.

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Influence of $C_5$-Precursors on $\delta$-Aminolevlinic Acid Biosynthesis in Rhodocyclus gelatinosus KUP-74 (Rhodocyclus gelatinosus KUP-74에 의한 $\delta$-Aminolevulinic acid 생합성의 $C_5$-전구물질의 영향)

  • 최경민;임왕진;황세영
    • Microbiology and Biotechnology Letters
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    • v.21 no.6
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    • pp.527-533
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    • 1993
  • Aminolevulinic acid(ALA) was shown to be synthesized via active pathways of either C4 or C5 ALA biosynthesis in cells of a photosynthetic bacterium, Rhodocyclus gelatinosus KUP-74, where the C5 pathway was appeared to be preferntially expressed in the cells. It was strongly suggested that L-glutamine might be utilized more effectively than L-glutamate to synthesize ALA via C5 pathway in this bacterium from the fact of relationship between the cellular uptake rates of glutamate and its Gamma-derivaties and corresponded ALA productivities in vitro and in vivo.

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Chemical Compositions & Texture of Allium senescens (두메부추의 화학적 조성 및 조직감 특성)

  • 정미숙;이미순
    • Korean journal of food and cookery science
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    • v.17 no.1
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    • pp.60-64
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    • 2001
  • Chemical compositions and texture of Allium senescens that is one of Korean indigenous culinary herbs were analyzed. The contents of carbohydrate and calorie were higher than those of Chinese chive. Proline, glutamine, valine, leucine and threonine were the major total amino acids in white and green portions of Allium senescens leaves. Methionine and cysteine were identified in white and green portions of Allium senescens leaves. The values of sodium and iron in Allium senescens were lower than in Chinese chive. Trace amount of selenium was also detected in Allium senescens leaves. The C$_{20:5}$ and C$_{20:6}$ were identified in leaves of Allium senescens. As a result of Rheometer analysis and sensory evaluation, texture of Allium senescens was more tender than that of green portion of Chinese chive leaves.

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Efficient plant regeneration from immature embryo cultures of Jatropha curcas, a biodiesel plant

  • Varshney, Alok;Johnson, T. Sudhakar
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.139-148
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    • 2010
  • Jatropha curcas L. (Physic nut) is a commercially important non-edible oil seed crop known for its use as an alternate source of biodiesel. In order to investigate the morphogenic potential of immature embryo, explants from four developmental stages were cultured on medium supplemented with combinations of auxins and cytokinins. It was found that the size of embryo is critical for the establishment of callus. Immature embryos (1.1-1.5 cm) obtained from the fruits 6 weeks after pollination showed a good response of morphogenic callus induction (85.7%) and subsequent plant regeneration (70%) with the maximum number of plantlets (4.7/explant) on Murashige and Skoog's (MS) medium supplemented with IBA (0.5 $mg\;l^{-1}$) and BA (1.0 $mg\;l^{-1}$). The above medium when supplemented with growth adjuvants such as 100 $mg\;l^{-1}$ casein hydrolysate + 200 $mg\;l^{-1}$ L-glutamine + 8.0 $mg\;l^{-1}$ $CuSO_4$ resulted in an even higher frequency of callus induction (100%). Plant regeneration (90%) with the maximum number of plantlets (10/explant) was achieved on MS medium supplemented with 500 $mg\;l^{-1}$ polyvinyl pyrrolidone + 30 $mg\;l^{-1}$ citric acid + 1 $mg\;l^{-1}$ BA + 0.5 $mg\;l^{-1}$ Kn + 0.25 $mg\;l^{-1}$ IBA. It was observed that plantlet regeneration could occur either through organogenesis of morphogenic callus or via multiplication of pre-existing meristem in immature embryos. The age of immature embryos and addition of a combination of growth adjuvants to the culture medium appear to be critical for obtaining high regeneration rates. Well-developed shoots rooted on half-halfstrength MS medium supplemented with 0.5 $mg\;l^{-1}$ IBA and 342 $mg\;l^{-1}$ trehalose. The rooted plants after acclimatization were successfully transferred to the field in different agro-climatic zones in India. This protocol has been successfully evaluated on five elite lines of J. curcas.