• Title/Summary/Keyword: Mass Identity

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Comparison of Anthocyanin Content in Seed Coats of Black Soybean [Glycine max(L.) Merr.] Cultivars Using Liquid Chromatography Coupled to Tandem Mass Spectrometry

  • Shin, Sung-Chul;Lee, Soo-Jung;Lee, Sung-Joong;Chung, Jong-Il;Bae, Dong-Won;Kim, Soo-Taek;Sung, Nak-Ju
    • Food Science and Biotechnology
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    • v.18 no.6
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    • pp.1470-1475
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    • 2009
  • The seed coat of the black soybean contains 3 main anthocyanins such as delphinidin-3-O-$\beta$-glucoside, cyanidin-3-O-$\beta$-glucoside, and petunidin-3-O-$\beta$-glucoside. As a part of our effort on discovering and breeding new black soybean cultivars which possesses specific anthocyanin component rich, we determined the anthocyanin profiles of the 2 cultivars recently developed soybean cv. Gaechuck #1 and cv. Gyeongsang #1, using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and compared their content and identity with those of previously known 10 cultivar controls. The Cosmosil-$5C_{18}$-AR-II column were selected for the analysis because of the best peak separation. The column temperature was set up at $35^{\circ}C$. The mobile phase consisting of water containing 0.5%(v/v) formic acid and methanol gave good separation between the 3 anthocyanin analytes and internal standard (quercetin 3-O-$\beta$-rutinoside) and peaks with suppressed tail. The MS/MS spectra of each individual anthocyanin standard were detected in positive electron spray ionization (ESI) modes. It was disclosed that the anthocyanin contents of the soybean cv. Gaechuck#1 and cv. Gyeongsang#1 are roughly higher than those of the 10 controls.

Functional Identification and Expression of Indole-3-Pyruvate Decarboxylase from Paenibacillus polymyxa E681

  • Phi, Quyet-Tien;Park, Yu-Mi;Ryu, Choong-Min;Park, Seung-Hwan;Ghim, Sa-Youl
    • Journal of Microbiology and Biotechnology
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    • v.18 no.7
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    • pp.1235-1244
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    • 2008
  • Indole-3-acetic acid (IAA) is produced commonly by plants and many bacteria, however, little is known about the genetic basis involving the key enzymes of IAA biosynthetic pathways from Bacillus spp. IAA intermediates from the Gram-positive spore-forming bacterium Paenibacillus polymyxa E681 were investigated, which showed the existence of only an indole-3-pyruvic acid (IPA) pathway for IAA biosynthesis from the bacterium. Four open reading frames (ORFs) encoding indole-3-pyruvate decarboxylase-like proteins and putative indole-3-pyruvate decarboxylase (IPDC), a key enzyme in the IPA synthetic pathway, were found on the genome sequence database of P. polymyxa and cloned in Escherichia coli DH5$\alpha$. One of the ORFs, PP2_01257, was assigned as probable indole-3-pyruvate decarboxylase. The ORF consisted of 1,743 nucleotides encoding 581 amino acids with a deduced molecular mass of 63,380 Da. Alignment studies of the deduced amino acid sequence of the ORF with known IPDC sequences revealed conservation of several amino acids in PP2_01257, essential for substrate and cofactor binding. Recombinant protein, gene product of the ORF PP2_01257 from P. polymyxa E681, was expressed in E. coli BL21 (DE3) as a glutathione S-transferase (GST)-fusion protein and purified to homogeneity using affinity chromatography. The molecular mass of the purified enzyme showed about 63 kDa, corresponding closely to the expected molecular mass of IPDC. The indole-3-pyruvate decarboxylase activity of the recombinant protein, detected by HPLC, using IPA substrate in the enzyme reaction confirmed the identity and functionality of the enzyme IPDC from the E681 strain.

Structural Characteristics of the Putative Protein Encoded by Arabidopsis AtMTN3 Gene

  • Cheong, Jong-Joo;Kwon, Hawk-Bin;Kim, Minkyun
    • Journal of Applied Biological Chemistry
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    • v.44 no.3
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    • pp.125-130
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    • 2001
  • A putative protein encoded by Arabidopsis AtMTN3 gene, a homologue of Medicago truncatula MTN3, consists of 285 amino acid residues, and has a predicted molecular mass of 31.5 kDa and a calculated pI of 9.1. Primary amino acid sequence analyses have revealed that the protein contains seven putative transmembrane regions with N-terminus oriented to the outside of the membrane. The AtMTN3 protein shows overall 16.4% of amino acid identity with the rat GALR3 protein, known to be a G-protein-coupled receptor. The gene is present as a single copy in the Arabidopsis genome, and expressed in aerial parts but not in roots of Arabidopsis. Therefore, AtMTN3 appears not to be specifically involved in Rhizobium-induced nodule development, as was predicted for the MTN3 gene. These proteins possibly mediate signal transmission through G-protein-coupled pathways during general interactions between plants and symbiotic or pathogenic microbes.

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Cloning and Expression of a Farnesyl Diphosphate Synthase in Centella asiatica (L.) Urban

  • Kim, Ok Tae;Ahn, Jun Cheul;Hwang, Sung Jin;Hwang, Baik
    • Molecules and Cells
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    • v.19 no.2
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    • pp.294-299
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    • 2005
  • A cDNA encoding farnesyl diphosphate synthase (FPS; EC2.5.1.1/EC2.5.1.10) was isolated from Centella asiacita (L.) Urban, using degenerate primers based on two highly conserved domains. A full-length cDNA clone was subsequently isolated by rapid amplification of cDNA ends (RACE) PCR. The sequence of the CaFPS (C. asiatica farnesyl diphosphate synthase) cDNA contains an open reading frame of 1029 nucleotides encoding 343 amino acids with a molecular mass of 39.6 kDa. The deduced CaFPS amino acid sequence exhibits 84, 79, and 72%, identity to the FPSs of Artemisia annua, Arabidopsis thaliana, and Oryza sativa, respectively. Southern blot analysis suggested that the C. asiatica genome contains only one FPS gene. An artificially expressed soluble form of the CaFPS was identified by SDS-PAGE. It had high specific activity and produced farnesyl diphosphate as the major isoprenoid.

Multidrug-resistant Staphylococci Found on Book Surfaces in East London Libraries

  • Idris, Adi;Cutler, Ron R
    • Microbiology and Biotechnology Letters
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    • v.45 no.4
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    • pp.354-357
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    • 2017
  • There is an increase in the presence of drug-resistant staphylococci outside of the nosocomial and healthcare setting. Although the presence of staphylococci has been studied in several public spaces, nothing is known on the presence of staphylococci in public libraries. Book surfaces from public libraries in the East London area, United Kingdom were swabbed and cultured and identity of the isolates determined by matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS). Seven different staphylococcal species were identified by MALDI-TOF-MS analysis. This short study provides evidence of the presence of multidrug-resistant staphylococci in public libraries in the East London area.

The Effect of Ethnic Identification and Social Group Affiliation on Body Image Satisfaction among Asian-American College Students

  • Lee, Yoon-Jung
    • International Journal of Human Ecology
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    • v.8 no.1
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    • pp.9-18
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    • 2007
  • This study focuses on the relationship between ethnic identity of Asian-Americans and their appreciation of their ethnic body features, based on reference group theory. A convenience sample of 60 male and 52 female students from various Asian ethnicities attending a mid-western university was used for the study. A 2 (gender) by 2 (ethnic identification) by 2 (socializing group) analysis of covariance (ANCOVA) on body image satisfaction as dependent variable and Body Mass Index score as a covariate was conducted. A significant main effect of ethnic identification was found, which indicates the more respondents identified with their ethnic group, the more likely they were to be satisfied with their appearance. The social group affiliation main effect was not significant. The impact of ethnic identification was significant only for those respondents who socialize more with Americans than with Asians. The results support the idea that one's ethnic group functions as a reference group, which influences body image appraisals.

N-Nitrosamine in Korean Beer

  • Sung, Nak-Ju;Shin, Jung-Hye;Lee, Soo-Jung
    • Preventive Nutrition and Food Science
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    • v.1 no.1
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    • pp.6-9
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    • 1996
  • A total of 47 beer samples, produced in Korea. Were analyzed for their N-nitrosamine levels by gas chromatography-thermal energy analyzer. N-nitrosodimethyamine(NDMA) was detected in 35 of 47 sam-ples analyzed. The range of levels was 0~7.2$\mu\textrm{g}$/kg while the average was about 0.9mg/kg. The identity of NDMA was confirmed by mass spectrometry. Samples which were found only NDMA before nitrosation were detected to include N-nitrosopyrroidine(45$\mu\textrm{g}$.kg) and N-nitrosomorpholine(4$\mu\textrm{g}$/kg) as well as NDMA(837$\mu\textrm{g}$/kg)

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Molecular Cloning of a Gene Encoding a Putative Antibacterial Peptide from Bombyx mori (누에에서 새로운 항세균성 펩타이드 유사 유전자의 분리와 염기서열 결정)

  • 김상현;제연호;윤은영;강석우;김근영;강석권
    • Korean journal of applied entomology
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    • v.35 no.4
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    • pp.321-325
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    • 1996
  • To isolate a novel gene for antibacterial peptide, an inducible clone(BmInc8) was selected by differential screening strategy from Bombyx mori cDNA library prepared from lavae injected with Escherichia coli. This clone contained a cDNA insert of 564 nucleotides and encoded 59 amino acids with an apparent molecular mass of 6.3 kDa. The cDNA sequence of BmInc8 had 61.2% identity compared to that of the bactericidin from Manduca sexta and also the deduced amino acids sequences from this insert had 65% identity compared to that of the cecropin D peptide Hyalophora cecropia. The transient expression assay of this insert using prokaryotic expression vector system revealed that the expressed peptide displayed the antibacterial activity. The cDNA sequence was deposited in GenBank under the accession number U30289.

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Molecular Cloning and Expression of a Novel Cuticle Protein Gene from the Chinese Oak Silkmoth, Antheraea pernyi

  • Kim Bo Yeon;Park Nam Sook;Jin Byung Rae;Kang Pil Don;Lee Bong Hee;Seong Su Il;Hwang Jae Sam;Chang Jong Su;Lee Sang Mong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.10 no.1
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    • pp.11-17
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    • 2005
  • In our research to identify gene involved in the cuticle protein, we cloned a novel cuticle protein gene, ApCP15.5, from the Chinese oak silkmoth, Antheraea pernyi, larvae cDNA library. The gene encodes a 149 amino acid polypeptide with a predicted molecular mass of 15.5 kDa and a pI of 9.54. The ApCP15.5 contained a type-specific consensus sequence identifiable in other insect cuticle proteins and the deduced amino acid sequence of the ApCP15.5 cDNA is most homologous to Tenebrio molitor-C1B ($43\%$ protein sequence identity), followed by Locusta migratoria-76 ($42\%$ protein sequence identity). Northern blot and Western blot analyses revealed that the ApCP15.5 showed the epidermis-specific expression. The expression profile of ApCP15.5 indicated that the ApCP15.5 mRNA expression was detected in the early stages after larval ecdysis and larval-pupal metamorphosis, and its expression level was most significant on the first day of larval ecdysis and pupal stage. The ApCP15.5 was expressed as a 15.5 kDa polypeptide in baculovirus-infected insect cells.

Isolation, Identification and Characterization of Phytophthora katsurae, Causing Chestnut Ink Disease in Korea

  • Lee, Jong-Kyu;Jo, Jong-Won;Shin, Keum-Chul;Lee, Sang-Hyun;Lee, Sang-Yong
    • The Plant Pathology Journal
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    • v.25 no.2
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    • pp.121-127
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    • 2009
  • Since July 2005, survey of chestnut ink disease was carried out in chestnut stands located at southern parts of Korea. Dead chestnut trees showing inky ooze on necrotic trunks were found in two different locations. In order to isolate and identify the causal fungus, infected tissues and soil samples around dead or dying trees were collected and placed on Phytophthora-selective medium. Rhododendron and chestnut tree leaves were used as a bait to isolate the fungus from soil samples by attracting zoospores in soil suspensions. On V-8 culture medium, the isolates produced homothallic oogonia with protuberances ($34.0-46.2{\times}21.9-26.7{\mu}m$) abundantly, but did not produced sporangia. Mass production of sporangia was possible by immersing agar plugs with actively growing mycelium in the creek water at $18^{\circ}C$ for 3 days. Sporangia were papillate, and ovoid to obpyriform ($17.0-38.9{\times}14.6-29.2{\mu}m$) in shape. Comparison of the ITS sequences revealed that the isolates had 100% identity to the P. katsurae isolates from Japan and New Zealand and 99.6% identity to other P. katsurae isolates. All of the examined isolates from Korea were completely identical to each other in ITS sequence. Numerous sporangia were formed in filtered as well as unfiltered creek water, but no sporangia formed in sterilized distilled water. Light induced sporangia formation, but has no influence on oospore formation. Amendments of ${\beta}$-sitosterol in culture media have no significant effect on mycelial growth but significantly stimulate oospore and sporangia formation.