• Title/Summary/Keyword: GLC analysis

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Functional Characterization and Application of the HpOCH2 Gene, Encoding an Initiating $\alpha$l,6-Mannosyltransferase, for N-glycan Engineering in the Methylotrophic Yeast Hansenula polymorpha

  • Kim, Moo-Woong;Kim, Eun-Jung;Kim, Jeong-Yoon;Rhee, Sang-Ki;Kang, Hyun-Ah
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2004.06a
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    • pp.278-281
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    • 2004
  • The $\alpha$1,6-mannosyltransferase encoded by Saccharomyces cerevisiae OCH1 plays a key role for the outer chain initiation of the N-linked oligosaccharides. A search for Hansenula polymorpha genes homologous to S. cerevisiae OCHI (ScOCH1) has revealed seven open reading frames (ORF100, ORF142, ORF168, ORF288, ORF379, ORF576, ORF580). All of the seven ORFs are predicted to be a type II integral membrane protein containing a transmembrane domain near the amino-terminal region and has a DXD motif, which has been found in the active site of many glycosyltransferases. Among this seven-membered OCH1 gene family of H. polymorpha, we have carried out a functional analysis of H. polymorpha ORF168 (HpOCH2) showing the highest identity to ScOCH1. Inactivation of this protein by disruption of corresponding gene resulted in several phenotypes suggestive of cell wall defects, including hypersensitivity to hygromycin B and sodium deoxycholate. The structural analysis of N-glycans synthesized in HpOCH2-disrupted strain (Hpoch2Δ) and the in vitro $\alpha$1,6-mannosyltransferase activity assay strongly indicate that HpOch2p is a key enzyme adding the first $\alpha$1,6-mannose residue on the core glycan Man$_{8}$GlcNAc$_2$. The Hpoch2Δ was further genetically engineered to synthesize a recombinant glycoprotein with the human compatible N-linked oligosaccharide, Man$_{5}$GlcNAc$_2$, by overexpression of the Aspergillus saitoi $\alpha$1,2-mannosidase with the 'HDEL” ER retention signal.gnal.

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A Study on Korean Green Tea(II) -Analysis of free Amino Acid and Mineral- (한국산(韓國産) 녹다(緣茶)에 대(對)한 연구(硏究) (제 II 보) -Free amino Acid와 무기성분(無機成分)에 대(對)하여-)

  • Jung, Jai-Kie;Yu, Choon-Hie;Chung, Tai-Young;La, Sang-Moo
    • Journal of Nutrition and Health
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    • v.6 no.3
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    • pp.17-20
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    • 1973
  • Quantitative analysis was achieved by gas-liquid chromatographic method (GLC) with a single column system of OV-17 for 16 of free amino acids in Korean green tea and the contents of mineral in it was determined by atomic absorption flame emission. The results are summarized as follows: 1) Korean green tea contained Mn 1% or over out of the total ash content and $0.05{\sim}0.20%$ in the water extraction, as the major mineral. 2) Ba, Cr, Ni, Pb and V were analyzed also by small quantities relatively and Co, Tin and Ywere not detected in the water extraction. 3) GLC indicated the presence of 16 components in free ammo acids. 4) The quantities of free amino acids in Korean green tea were determined $2.96{\sim}6.61%$ Alanine, $1.01{\sim}l.89mg%$ Glycine, $2.07{\sim}7.81mg%$ Valine, $1.27{\sim}8.76mg%$ Leucine and Isoleucine, $94.31{\sim}316.27mg%$ Threonine, $9.10{\sim}39.91mg%$ Serine, $2.18{\sim}36.76mg%$ Hydroxyproline, $2.72{\sim}5.90mg%$ Proline, $39.64{\sim}70.02mg%$ Aspartic Acid, $25.93{\sim}101.28mg%$ Glutamic Acid and Lysine, $8.32{\sim}18.30mg%$ Phenylalanine and Tyrosine in trace amount. 5) The total free amino acid contents in Korean green tea ranged from 207.24mg% to 516.06mg% and Moo-Deoung tea contained outstandingly high, 516 mg% or over.

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Studies on Analysis of Sterols in Mayonnaise by GLC with Packed and Capillary Column (충전컬럼 및 모세관컬럼 가스크로마토그라프에 의한 마요네즈 중의 스테롤 분석연구)

  • Kim, Hyean-Wee;Jeong, So-Young;Jeong, Chang-Ki;Yoon, Hyeong-Sik;Park, Ki-Moon;Ahn, Peong-Ug;Choi, Chun-Un
    • Korean Journal of Food Science and Technology
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    • v.24 no.1
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    • pp.82-85
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    • 1992
  • Sterols in mayonnaise were determined by the gas liquid chromatographic(GLC) analysis using two different columns, packed column(silicone SE30) and fused silica capillary column(CBPl). Four kinds of sterol(cholesterol, campesterol, stigmasterol and ${\beta}-sitosterol$) were detected in mayonnaise. The method using capillary column proved to be superior in rapidity, reproducerbility and separability for the determination of sterols. The ranges of concentration of egg york in mayonnaise predicted from cholesterol content using packed and capillary columns were $2.69{\sim}7.11%\;and\;2.42{\sim}6.08%$, respectively. Analyzing the composition of campesterol, stigmasterol and (${\beta}-sitosterol$ in mayonnaise, it could be known that soybean oil and cottonseed oil are commonly used for the manufacture of mayonnaise.

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Correlation of saponarin content with biosynthesis-related gene expression in hulled and hulless barley (Hordeum vulgare L.) cultivars

  • Lee, HanGyeol;Park, Jae-Hyeok;Yoon, A Mi;Kim, Young-Cheon;Park, Chul Soo;Yang, Ji Yeong;Woo, So-Yeun;Seo, Woo Duck;Lee, Jeong Hwan
    • Journal of Plant Biotechnology
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    • v.48 no.1
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    • pp.12-17
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    • 2021
  • Saponarin found in young barley sprouts has a variety of beneficial biological and pharmacological properties, including antioxidant, hypoglycemic, antimicrobial, and hepatoprotective activities. Our previous work demonstrated that saponarin content was correlated with the expression levels of three biosynthetic pathway genes [chalcone synthase (HvCHS1), chalcone isomerase (HvCHI), and UDP-Glc:isovitexin 7-O-glucosyltransferase (HvOGT1)] in young barley seedlings under various abiotic stress conditions. In this study, we investigated the saponarin content and expression levels of three saponarin biosynthetic pathway genes in hulled and hulless domestic barley cultivars. In the early developmental stages, some hulled barley cultivars (Kunalbori1 and Heukdahyang) had much higher saponarin contents than did the hulless barley cultivars. An RNA expression analysis showed that in most barley cultivars, decreased saponarin content correlated with reduced expression of HvCHS1 and HvCHI, but not HvOGT1. Heat map analysis revealed both specific increases in HvCHS1 expression in certain hulled and hulless barley cultivars, as well as general changes that occurred during the different developmental stages of each barley cultivar. In summary, our results provide a molecular genetic basis for the metabolic engineering of barley plants to enhance their saponarin content.

Studies on the Triglyceride Composition of Some Vegetable Oils -IV. Triglyceride Composition of Rice Bran Oil- (식물유(植物油)의 Triglyceride조성(組成)에 관한 연구(硏究) -제(第) 4 보(報) : 쌀겨기름의 Triglyceride조성(組成)-)

  • Choi, Su-An;Park, Yeung-Ho
    • Korean Journal of Food Science and Technology
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    • v.15 no.2
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    • pp.108-111
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    • 1983
  • High performance liquid chromatography (HPLC) was applied to the analysis of triglycerides of rice bran oil. The triglycerides were clearly separated in five peaks by HPLC on a column packed with ${\mu}-Bondapack$ C18 using methanol-chloroform mixture as a solvent. Compositions of the triglyceride and fatty acid of the fraction was also analyzed by gas liquid chromatography (GLC). Each of these collected fractions gave three to four peaks in the GLC chromatograms according to the carbon number of the triglyceride. The fitty acid compositions of these triglycerides were mainly composed of C16:0, C18:1 and C18:2 fatty acids. The major triglycerides of the rice bran oil were found to be those of (C16:0, C18:1, C18:2;16.64%), $(2{\times}C18:1,\;C18:2;16.18%)$, $(3{\times}C18:1;13.7%)$, $(C16:0,\;2{\times}C18:1;12.77%)$,$(C18:1,\;2{\times}C18:2;9.16%)$ and $(C16:0,\;2{\times}C18:2;6.42%)$

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Tandem Mass Spectrometry of N-linked Glycans from Human Immunoglobulin G (다중 질량 분석법을 이용한 인체 면역글로불린 G의 N-연결 글라이칸 분석)

  • Joo, Hwang-Soo;Kim, Yun-Gon;Jang, Kyoung-Soon;Kim, Byung-Gee
    • KSBB Journal
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    • v.22 no.4
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    • pp.234-238
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    • 2007
  • We used electrospary ionization ion trap tandem mass spectrometry (ESI-IT tandem MS) to structural elucidation of three different biantennary-type glycans having zero, one, two galactoses (G0, G1, G2). The highest fragment ion in the MS/MS spectra of three glycans was produced by 0,2-ring cleavage of fucose-linked N-acetylglucosamine (GlcNAc) in reducing end. The fragment ions both from precursor ions and 0,2-ring cleaved ions ($^{0.2}An$; n=5 for G0, n=6 for G1 and G2) were not overlapped each other. As results of $MS^n$ analyses, tandem fragmentation trees of each glycans were generated and 2,4-ring cleavages ($^{2.4}A_6$) were occurred in GlcNAc linked to reducing end GlcNAc. This structural elucidation and fragmentation study of N-linked glycans by tandem mass spectrometry can be applied to structural analysis of more complicated glycans.

Lipids and Fatty Acid Composition of Herba Capsellae (Capgella bursa-Pastoris Linne) (냉이의 지방질(脂肪質) 및 지방산(脂肪酸) 조성에 관한 연구(硏究))

  • Bae, Man-Jong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.16 no.2
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    • pp.83-86
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    • 1987
  • Lipids extracted and purified from Herbs Capsellae leaf ana root were studied, To investigate those, silicic acid column chromatography, TLC, GLC was used for analysis of lipids and Fatty acids composition. Following results were obtained. 1. From result of total lipid classes in Herba Capsellae leaf and root, NL were obtained 62.6%, 58.5% respectly. GL 20.8%, 17,8%. PL 16.6%, 23.7%, respectively. 2. Among the NL in Herba capsellae leaf and root, TG were predominant 77.5%, 80.3%, respectly. SE 9.2%, 8.4%. FFA 4.8%, 3.4%. 1.2-DG 4.1%, 3.6%. FS 2.2%, 2.2%. and 1, 3-DG 1.3%, 1.0%, in turn. 3. The major fatty acids in the lipid classes were linoic acid, palmitic acid, oleid acid, in all. And total unsaturated fatty acid was NL 78.0%, GL 66.6%, and PL 75.2% in Herbs capsellae leaf, and NL 66.1% GL 63.6% and PL 61.2% in Herbs capsellae root.

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Evaluation of Organochlorine Pesticide Residues in Soil by Steam Distillation (수증기(水蒸氣) 증류법(蒸溜法)에 의(依)한 토양중(土壤中) 유기염소계(有機鹽素系) 살충제의 잔류분(殘留分) 평가(評價))

  • Suh, Yong-Tack;Shim, Jae-Han;Park, Ro-Dong
    • Korean Journal of Environmental Agriculture
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    • v.3 no.2
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    • pp.23-29
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    • 1984
  • Agricultural soil samples collected from 121 sites (plastic film house 59, up-land 30 and orchard 32 sites) were evaluated by steam distillation and GLC-ECD analysis for organochlorine pesticides (${\gamma}-BHC$, heptachlor, p,p'-DDE, dieldrin, p,p'-DDD, and p,p'-DDT) in Jeollanam-Do area. Total residues of organochlorine pesticide in orchard, plastic film house and up-land were 0.415, 0.234 and 0.156ppm, respectively. Ninety-four percent of total residue was p,p'-DDT and its homologues (p,p'-DDE and p,p'-DDD). ${\gamma}-BHC$ was detected in all soil samples at $trace{\sim}0.050ppm$ range. Residue levels of organochlorine pesticide increased in the order of p,p'-DDE, ${\gamma}-BHC$, heptachlor, dieldrin, p,p'-DDT and p,p'-DDD.

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Studies on the Residues of Chlorinated Organic Insecticides -2. BHC Residues in or on Commercial Vegetables- (유기염소계(有機鹽素系) 살충제(殺蟲劑)의 잔유분(殘留分)에 관(關)한 연구(硏究) -2. 시판채소중(市販菜蔬中) BHC 잔유분(殘留分)에 관(關)하여-)

  • Park, Chang-Kyu;Lee, Kyu-Seung;Yoo, Jai-Yoo
    • Applied Biological Chemistry
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    • v.17 no.3
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    • pp.177-183
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    • 1974
  • One hundred and twenty-five samples comprising thirteen vegetables harvested in 1971 were collected and subjected to GLC analysis, for ${\alpha}-and\;{\gamma}-BHC$ residues, using ECD. Heavy dependence on the technical BHC, instead of Lindane, was amply reflected on the BHC residues-residues of ${\alpha}-BHC$, depending on the vegetables, amounted to $2{\sim}7$ times that of ${\gamma}-BHC$. In spite of extensive contamination by the BHC isomers, residue levels of ${\gamma}-BHC$ were less than 0.015 ppm in all vegetables and were far less than the tolerence limit of $0{\sim}3.0ppm$ recommended jointly by the FAO and WHO.

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Applications of Tandem Mass Spectrometry in the Structure Determination of Permethylated Sialic Acid-containing Oligosaccharides

  • Yoo, Eun-Sun;Yoon, In-Mo
    • Bulletin of the Korean Chemical Society
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    • v.26 no.9
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    • pp.1347-1353
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    • 2005
  • Sets of sialic acid-containing trisaccharides having different internal and terminal linkages have been synthesized to develop a sensitive method for analysis of the reducing terminal linkage positions. The trisaccharides, sialyl($\alpha$ 2-3)Gal($\beta$ 1-3)GalNAc and sialyl($\alpha$ 2-3)Gal($\beta$ 1-X)GlcNAc where X=3, 4 and 6, were synthesized and examined using electrospray ionization (ESI)-collision induced dissociation (CID) tandem mass spectrometry (MS/MS). The compounds chosen for this study are related to terminal groups likely to be found on polylactosamine-like glycoproteins and glycolipids which occur on the surface of mammalian cells. The purpose of this study is to develop tandem mass spectrometral methods to determine detailed carbohydrate structures on permethylated or partially methylated oligosaccharides for future applications on biologically active glycoconjugates and to exploit a faster method of synthesizing a series of structural isomeric oligosaccharides to be used for further mass spectrometry and instrumental analysis.