• Title/Summary/Keyword: L-glutamine

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T-Cell Death-Associated Gene 51 Is a Novel Negative Regulator of PPARγ That Inhibits PPARγ-RXRα Heterodimer Formation in Adipogenesis

  • Kim, Sumi;Lee, Nari;Park, Eui-Soon;Yun, Hyeongseok;Ha, Tae-Uk;Jeon, Hyoeun;Yu, Jiyeon;Choi, Seunga;Shin, Bongjin;Yu, Jungeun;Rhee, Sang Dal;Choi, Yongwon;Rho, Jaerang
    • Molecules and Cells
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    • v.44 no.1
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    • pp.1-12
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    • 2021
  • The nuclear receptor peroxisome proliferator-activated receptor γ (PPARγ) is the master transcriptional regulator in adipogenesis. PPARγ forms a heterodimer with another nuclear receptor, retinoid X receptor (RXR), to form an active transcriptional complex, and their transcriptional activity is tightly regulated by the association with either coactivators or corepressors. In this study, we identified T-cell death-associated gene 51 (TDAG51) as a novel corepressor of PPARγ-mediated transcriptional regulation. We showed that TDAG51 expression is abundantly maintained in the early stage of adipogenic differentiation. Forced expression of TDAG51 inhibited adipocyte differentiation in 3T3-L1 cells. We found that TDAG51 physically interacts with PPARγ in a ligand-independent manner. In deletion mutant analyses, large portions of the TDAG51 domains, including the pleckstrin homology-like, glutamine repeat and proline-glutamine repeat domains but not the proline-histidine repeat domain, are involved in the interaction with the region between residues 140 and 506, including the DNA binding domain, hinge, ligand binding domain and activation function-2 domain, in PPARγ. The heterodimer formation of PPARγ-RXRα was competitively inhibited in a ligand-independent manner by TDAG51 binding to PPARγ. Thus, our data suggest that TDAG51, which could determine adipogenic cell fate, acts as a novel negative regulator of PPARγ by blocking RXRα recruitment to the PPARγ-RXRα heterodimer complex in adipogenesis.

Analysis of Mineral, Amino Acid and Vitamin Contents of Fruiting Body of Sparassis crispa. (꽃송이버섯의 미네랄, 아미노산, 비타민 함량분석)

  • Shin, Hyun-Jae;Oh, Deuk-Sil;Lee, Hee-Duck;Kang, Hyeong-Bong;Lee, Chul-Won;Cha, Wol-Suk
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1290-1293
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    • 2007
  • The nutritional composition of fruiting body of Sparassis crispa has been analyzed for medicinal and edible uses. Minerals in S. crispa were found to be as follows; potassium (1,299.44 mg), phosphorus (104.73 mg), sodium (98.21 mg), magnesium (54.86 mg), calcium (8.39 mg), iron (7.61 mg), zinc (6.37 mg), copper (1.31 mg) and manganese (0.63 mg) based on 100 g of mushroom dry weight. In 20 kinds of total amino acids found in S. crispa, sum of glutamic acid and glutamine content was the highest (1,960 mg/l00 g) and sum of aspartic acid and asparagine, tryptophan, leucine and alanine were followed. Concerning free amino acids, glutamic acid, tryptophan, glutamine and aspartic acid were dominant. Among 8 vitamins detected, the vitamin E content was the highest (408.5 mg) based on 100 g of mushroom dry weight, then vitamin C, niacin and pantothenic acid were followed.

Sulfur Deficiency Effects on Nitrate Uptake and Assimilatory Enzyme Activities in Rape Plants (유채에서 황 결핍이 질산염의 흡수 및 동화관련 효소활력에 미치는 영향)

  • Li, Lu-Shen;Jin, Yu-Lan;Lee, Bok-Rye;Kim, Tae-Hwan
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.29 no.2
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    • pp.103-110
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    • 2009
  • Rape plants, especially forage type, are known as one of high S-demanding plants. Their productivity and quality have often limited under S-deficient condition. To investigate the effect of S-deficiency on N uptake and its assimilation, $NO_3^-$ absorption, nitrate reducatse (NR) and glutamine synthetase (GS) activity in leaf and root tissues as affected by different S-supplied level was determined. $NO_3^-$ uptake was not significant between control and S-deficient treated plants, while significantly depressed in S-deprived plants for the early 8 h. NR activity decreased as S-availability decreased, especially in young and middle leaves, representing more than 35% of decrease in S-deficient and 70% in S-deprived plants when compared with control. In roots, a significant decrease (-29%) in NR was observed only in S-deprived plants. Relatively higher GS activity was found in young leaves for three all treatments. As a whole leaf tissue, S-limited conditions resulted in a reduction of GS activity. In root which showed the lowest activity, a significant decrease (-30%) was observed only in S-deprived plants.

Changes of Nitrogen Uptake, Growth and Activities of Nitrogen Metabolizing Enzymes by Different Source of Nitrogen in Tobacco (담배에서 질소 형태에 따른 흡수 양상 및 생육과 질소대사 효소의 활성 변화)

  • 이상각;심상인;강병화;배길관
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.42 no.5
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    • pp.515-521
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    • 1997
  • Tobacco plant was grown for 40 days hydroponically in nutrient solutions composed of different forms of nitrogen, like NO$_3$$^{[-10]}$ -N, NH$_4$$^{+}$-N, and a mixed formulation of NO$_3$$^{[-10]}$ -N and NH$_4$$^{+}$-N. Uptake response, nitrate reductase, and glutamine synthetase activity at growth stage were investigated to understand the basic knowledge of nitrogen metabolism. The better growth of shoot and root was observed in the mixed nutrient solution than NO$_3$$^{[-10]}$ -N or NH$_4$$^{+}$-N, alone. The plant growth in NH$_4$$^{+}$-N nutrient solution was poor due to ammonium toxicity. The pH of nutrient solution containing NO$_3$$^{[-10]}$ -N increased up to 40 days after transplanting. But the pH of solution containing NO$_3$$^{[-10]}$ -N decreased drastically to 3.42 at 20 days after transplant. The pH in the mixed formulation dropped to pH 3.64 at 30 days after transplant and showed re-increase. It is assumed that nitrogen of NH$_4$$^{+}$-N form was taken up preferentially at early stage and NO$_3$$^{[-10]}$ -N form was taken up preferentially at middle stage in the treatment with the mixed solution. The result indicates that the relative proportion of nitrogen forms affected the uptake patterns at each growth stages. The contents of chlorophyll and soluble protein were high with the mixed solution. Total nitrogen content was the highest in NH$_4$$^{+}$-N solution and the content also increased by the application of the mixed type of nitrogen. The amount of nitrate in leaves was high in NO$_3$$^{[-10]}$ -N treatment and the amount of ammonium was high in NH$_4$$^{+}$-N treatment. The activity of nitrate reductase or glutamine synthetase was highest in the leaves grown in mixed nutrient solution than in those with any other single of nitrogen form.

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Selection of Lactococcus lactis HY7803 for Glutamic Acid Production Based on Comparative Genomic Analysis

  • Lee, Jungmin;Heo, Sojeong;Choi, Jihoon;Kim, Minsoo;Pyo, Eunji;Lee, Myounghee;Shin, Sangick;Lee, Jaehwan;Sim, Jaehun;Jeong, Do-Won
    • Journal of Microbiology and Biotechnology
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    • v.31 no.2
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    • pp.298-303
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    • 2021
  • Comparative genomic analysis was performed on eight species of lactic acid bacteria (LAB)-Lactococcus (L.) lactis, Lactobacillus (Lb.) plantarum, Lb. casei, Lb. brevis, Leuconostoc (Leu.) mesenteroides, Lb. fermentum, Lb. buchneri, and Lb. curvatus-to assess their glutamic acid production pathways. Glutamic acid is important for umami taste in foods. The only genes for glutamic acid production identified in the eight LAB were for conversion from glutamine in L. lactis and Leu. mesenteroides, and from glucose via citrate in L. lactis. Thus, L. lactis was considered to be potentially the best of the species for glutamic acid production. By biochemical analyses, L. lactis HY7803 was selected for glutamic acid production from among 17 L. lactis strains. Strain HY7803 produced 83.16 pmol/μl glutamic acid from glucose, and exogenous supplementation of citrate increased this to 108.42 pmol/μl. Including glutamic acid, strain HY7803 produced more of 10 free amino acids than L. lactis reference strains IL1403 and ATCC 7962 in the presence of exogenous citrate. The differences in the amino acid profiles of the strains were illuminated by principal component analysis. Our results indicate that L. lactis HY7803 may be a good starter strain for glutamic acid production.

Changes in Chemical Compositions of Pumpkin(Cucurbita moschata DUCH.) Seed Sprouts (호박(Cucurbita moschata DUCH.)종실의 발아 성장 과정 중 성분 변화)

  • 이병진;장희순;이규희;오만진
    • Food Science and Preservation
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    • v.10 no.4
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    • pp.527-533
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    • 2003
  • This study was performed for increasing the consumption and developing the function of pumpkin(Cucurbita moschata DUCH.) seed. The changes of the contents of general chemical compositions, fatty acids, amino acids, ascorbic acid and ${\beta}$-carotene during sprouting were analyzed. Also, the bitter taste, which was produced during sprouting, were purified by using thin layer chromatography and preparative high pressure liquid chromatography. The purified bitter compound was identified by mass spectrum and nuclear magnetic resonance($^1$H '||'&'||' $\^$13/C-NMR). Weight of pumpkin seed sprout was increased to 348.4% and the length of stem was dramatically increased at 8 days. In each head and stem parts of the pumpkin seed sprout, the contents of protein and lipid were decreased, however, the contents of fiber, ash and soluble inorganic nitrogen were increased. The fatty acids of the pumpkin seed sprout were mainly represented as linoleic acid, oleic acid, palmitic acid and stearic acid. During sprouting, palmitic acid was gradually increased, reversely, linoleic acid was gradually decreased. The general amino acids of head part in the pumpkin seed sprout grown at 23$^{\circ}C$ during 8 days were orderly more contained glycine, alanine, arginine, cystein and proline. Those of free amino acids were orderly more contained arginine, threonine, alanine and glutamine. The contents of L-ascorbic acid and ${\beta}$-camtene of the pumpkin seed sprout were gradually increased with increasing sprouting days. The bitter taste material of head part of the pumpkin seed sprout was detected at Rf value 0.72 on silicagel TLC plate and separuted as one peak by HPLC. The chemical structure of the puified bitter compound was identified as a cucurbitacin glycoside by MS and NMR. The content of bitter compound at 8 days was contained 42.2 mg per 1kg sprout head.

Development of Bialaphos-Resistant Transgenic Rice Using Agrobacterium tumefaciens (Agrobacterium tumefaciens를 이용한 bialaphos 저항성 형질전환 벼의 개발)

  • 이효연;이춘환;김호일;한원동;최지은;김진호;임용표
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.4
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    • pp.283-288
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    • 1998
  • The bialaphos is a potent inhibitor of glutamine synthease in higher plants and is used as a non-selective herbicide. We have used the bialaphos resistant gene(Bar) encoding for an acetyltransferase isolated from Streptomyces hygroscopicus SF1293. Callus derived from mature seeds of rice(Oryza sativa L. cv. Dong Jin) were co-cultivated with Agrobacterium tumefaciens EHA101 carring a plasmid pGPTV-HB containing genes for hygromycin resistance (HygR) and Bar. Transgenic plants showing in vitro resistance to 50 mg/L hygromycin and 10 mg/L bialaphos were obtained by using a two-step selection/regeneration procedure. Transformation efficiency of rice was about 30% which was as high as reported in other dicotyledons. Progenies ($\textrm{T}_{1}$ generation) derived from primary transformant of 17 lines were segregated with a 3 resistant : 1 sensitive ratio in medium containing hygromycin and bialaphos. Stable integration of Bar gene into chromosomal DNA was proven by Southern blot analysis of genomic DNA isolated from $\textrm{T}_{2}$ progenies. Transgenic plants ($\textrm{T}_{3}$) grown in the field were resistant to bialaphos (Basta) at a dosage lethal to wild type plants.

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Nutritional and Bioactive Compounds of Yellow Cherry Tomato (황색 방울토마토의 영양성분 및 생리활성 물질분석)

  • Choi, Suk-Hyun
    • The Journal of the Korea Contents Association
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    • v.20 no.2
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    • pp.451-461
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    • 2020
  • This study was carried out to investigate the value of yellow cherry tomato as a food material. Contents of nutritional and bioactive compound were determined from yellow cherry tomato. 17 of amino acids occurred. L-Glutamic acid (45.15%), L-glutamine (22%) and L-aspartic acid (11%) were the main amino acids. 8 of essential amino acids were found except tryptophan. γ-aminobutyric acid (GABA) was found in high contentration (258.58 mg/100g). Contents of Lycopene and β-carotene were 2.18 mg/100 g and 9.90 mg/100 g, respectively. Yellow cherry tomato contained naringenin chalcone, quercetin-3-rutinoside(rutin), 5-caffeoylquinic acid, 3-caffeoylquinic acid, and quercetin-3-apiosylrutinoside that have various bioactivities. These results revealed that yellow cherry tomato would be very useful and valuable food material.

Anticancer Activity of Extremely Effective Recombinant L-Asparaginase from Burkholderia pseudomallei

  • Darwesh, Doaa B.;Al-Awthan, Yahya S.;Elfaki, Imadeldin;Habib, Salem A.;Alnour, Tarig M.;Darwish, Ahmed B.;Youssef, Magdy M.
    • Journal of Microbiology and Biotechnology
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    • v.32 no.5
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    • pp.551-563
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    • 2022
  • L-asparaginase (E.C. 3.5.1.1) purified from bacterial cells is widely used in the food industry, as well as in the treatment of childhood acute lymphoblastic leukemia. In the present study, the Burkholderia pseudomallei L-asparaginase gene was cloned into the pGEX-2T DNA plasmid, expressed in E. coli BL21 (DE3) pLysS, and purified to homogeneity using Glutathione Sepharose chromatography with 7.26 purification fold and 16.01% recovery. The purified enzyme exhibited a molecular weight of ~33.6 kDa with SDS-PAGE and showed maximal activity at 50℃ and pH 8.0. It retained 95.1, 89.6%, and 70.2% initial activity after 60 min at 30℃, 40℃, and 50℃, respectively. The enzyme reserved its activity at 30℃ and 37℃ up to 24 h. The enzyme had optimum pH of 8 and reserved 50% activity up to 24 h. The recombinant enzyme showed the highest substrate specificity towards L-asparaginase substrate, while no detectable specificity was observed for L-glutamine, urea, and acrylamide at 10 mM concentration. THP-1, a human leukemia cell line, displayed significant morphological alterations after being treated with recombinant L-asparaginase and the IC50 of the purified enzyme was recorded as 0.8 IU. Furthermore, the purified recombinant Lasparaginase improved cytotoxicity in liver cancer HepG2 and breast cancer MCF-7 cell lines, with IC50 values of 1.53 and 18 IU, respectively.

Characterization of a Serine Protease from Neungee [Sarcodon aspratus(Berk, ) S. Ito] (능이[Sarcodon aspratus(Berk, ) S. Ito]에서 분리한 단백질 가수분해 효소의 특성)

  • 엄태붕;유관성;김미경;류재수;손희숙;이태규
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.20 no.1
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    • pp.35-39
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    • 1991
  • Properties of a protease purified from Neungee[Sarcodon aspratus(Berk, ) S. Ito] have been investigated. The enzyme displays a glycosylated serine protease. The enzyme is able to hydrolyze alanine glycine methionine glutamine and cysteine of N-CBZ and N-t-BOC-L-amino acid derivatibes relatively strongly but splits valine proline and isoleucine derivatives with low affinity which means the enzyme has the broad substrate spectrum toward the amino acids. Interestingly the enzyme was inhibited by bromelain inhibitor. That is the active site environ-ment of the enzyme is believed to be similar to that of bromelain However peptide mapping studies show that the two enzymes have distinct different cleavage sites.

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