• 제목/요약/키워드: sequence-based screening

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Molecular Diversity of Fungal Endophytes Isolated from Garcinia mangostana and Garcinia parvifolia

  • Sim, Jiun-Horng;Khoo, Chai-Hoon;Lee, Learn-Han;Cheah, Yoke-Kqueen
    • Journal of Microbiology and Biotechnology
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    • 제20권4호
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    • pp.651-658
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    • 2010
  • Garcinia is commonly found in Malaysia, but limited information is available regarding endophytic fungi associated with this plant. In this study, 24 endophytic fungi were successfully recovered from different parts of two Garcinia species. Characterization of endophytic fungi was performed based on the conserved internal transcribed spacer (ITS) region sequence analysis and the antimicrobial properties. Results revealed that fruits of the plant appeared to be the highest inhabitation site (38%) as compared with others. Glomerella sp., Guignardia sp., and Phomopsis sp. appeared to be the predominant endophytic fungi group in Garcinia mangostana and Garcinia parvifolia. Phylogenetic relationships of the isolated endophytic fungi were estimated from the sequences of the ITS region. On the other hand, antibacterial screening showed 11 of the isolates possessed positive response towards pathogenic and nonpathogenic bacteria. However, there was no direct association between certain antibacterial properties with the specific genus observed.

Potential Yeast from Indonesian Wild Forest Honey Showing Ability to Produce Lipase for Lipid Transesterification

  • Palilu, Prayolga Toban;Kasiamdari, Rina Sri;Ilmi, Miftahul
    • 한국미생물·생명공학회지
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    • 제47권4호
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    • pp.555-564
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    • 2019
  • Biodiesel is produced through the transesterification process in the presence of alcohol and a catalyst that catalyzes the conversion of triglycerides to esters and glycerol compounds. A more optimal product conversion can be achieved using enzymes, such as lipase. Lipase is reported to be produced in osmophilic yeasts due to the low water content in their natural habitats. Wild forest honey is one of the osmophilic natural habitats in Indonesia. However, lipase-producing yeast has not been reported in the Indonesian honey. In this study, we screened the lipase-producing yeasts isolated from wild forest honey collected from Central Sulawesi. The production profile and activity of lipase were determined at different pH values and temperatures. One promising yeast was isolated from the honey, which was identified as Zygosaccharomyces mellis SG 1.2 based on ITS sequence. The maximum lipase production (24.56 ± 1.30 U/mg biomass) was achieved by culturing the strain in a medium containing 2% olive oil as a carbon source at pH 7 and 30℃ for 40 h. The optimum pH and temperature for lipase activity were 6 and 55℃, respectively. The enzyme maintained 80% of its activity upon incubation at 25℃ for 4 h. However, the enzyme activity decreased by more than 50% upon incubation at 35 and 40℃ for 2 h. This is the first study to report the lipase producing capability of Z. mellis. Further studies are needed to optimize the enzyme production.

Genetic Variation in a DNA Double Strand Break Repair Gene in Saudi Population: A Comparative Study with Worldwide Ethnic Groups

  • Areeshi, Mohammed Yahya
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권12호
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    • pp.7091-7094
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    • 2013
  • DNA repair capacity is crucial in maintaining cellular functions and homeostasis. However, it can be altered based on DNA sequence variations in DNA repair genes and this may lead to the development of many diseases including malignancies. Identification of genetic polymorphisms responsible for reduced DNA repair capacity is necessary for better prevention. Homologous recombination (HR), a major double strand break repair pathway, plays a critical role in maintaining the genome stability. The present study was performed to determine the frequency of the HR gene XRCC3 Exon 7 (C18067T, rs861539) polymorphisms in Saudi Arabian population in comparison with epidemiological studies by "MEDLINE" search to equate with global populations. The variant allelic (T) frequency of XRCC3 (C>T) was found to be 39%. Our results suggest that frequency of XRCC3 (C>T) DNA repair gene exhibits distinctive patterns compared with the Saudi Arabian population and this might be attributed to ethnic variation. The present findings may help in high-risk screening of humans exposed to environmental carcinogens and cancer predisposition in different ethnic groups.

사람 티로시나제의 3차원 구조 상동 모델링 (Comparative modeling of human tyrosinase - An important target for developing skin whitening agents)

  • 최종근;서주원
    • 한국산학기술학회:학술대회논문집
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    • 한국산학기술학회 2012년도 춘계학술논문집 1부
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    • pp.182-186
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    • 2012
  • human tyrosinase (hTyr) catalyzes first and the rate limiting step in the synthesis of polymerized pigment, melanin which determines skin, hair and eye colors. Mutation of hTyr often brings about decrease of melanin production and further albinism. Meanwhile, a number of cosmetic companies providing skincare products for woman in Asia-Pacific region have tried to develop inhibitors to bright skin color for several decades. In this study, we built a 3D structure by comparative modeling technique based on the crystal structure of tyrosinase from bacillus megaterium as a template to serve structural information of hTyr. According to our model and sequence analysis of type 3 copper protein family proteins, two copper atoms of active site located deep inside are coordinated with six strictly conserved histidine residues coming from four-helix-bundle. Cavity which accommodates substrates was like funnel shape of which entrance was wide and expose to solvent. In addition, protein-substrate and protein-inhibitor complex were modeled with the guide of van der waals surface generated by in house software. Our model suggested that only phenol group or its analogs can fill the binding site near nuclear copper center because inside of binding site has narrow shape relatively. In conclusion, the results of this study may provide helpful information for designing and screening new anti-melanogensis agents.

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Characterization of a Lichenase Isolated from Soil Metagenome

  • Kim, Sang-Yoon;Oh, Doo-Byoung;Kwon, Ohsuk
    • Journal of Microbiology and Biotechnology
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    • 제24권12호
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    • pp.1699-1706
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    • 2014
  • A lichenase gene (mt-lic) was identified for the first time through function-based screening of a soil metagenomic library. Its deduced amino acid sequence exhibited a high degree of homology with endo-${\beta}$-1,3-1,4-glucanase (having both lichenase and chitosanase activities), encoded by the bgc gene of Bacillus circulans WL-12. The recombinant lichenase overexpressed and purified from Escherichia coli was able to efficiently hydrolyze both barley ${\beta}$-glucan and lichenan. The enzyme showed maximal activity at a pH of 6.0 at $50^{\circ}C$, with Azo-barley-glucan as the substrate. The metal ions $Mn^{2+}$, $Mg^{2+}$, $Ca^{2+}$, and $Fe^{2+}$ enhanced the enzymatic activity, whereas the $Cu^{2+}$ and $Zn^{2+}$ ions inhibited the enzymatic activity. The $K_m$ and $V_{max}$ values of the purified lichenase were determined to be 0.45 mg/ml and 24.83 U/min/mg of protein, respectively.

위해성평가와 관리를 고려한 유전자변형작물 개발의 필요성 (Development of genetically modified crops based on considerations of risk assessment and management)

  • 김창기;정순천;윤원기;박기웅;최경화;김환묵
    • Journal of Plant Biotechnology
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    • 제36권4호
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    • pp.360-365
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    • 2009
  • Over the last five years, we have conducted research on risk assessment of domestically developed genetically modified (GM) crops and found a number of factors which could delay risk assessment process. In this review, we described such cases and discussed the problem of transgene cassette integration, the lack of information on vectors, the poor quality control in seed production and absence of bioinformatic analysis on amino acid sequence homology before GM crop development. To solve these problems, we have suggested the introduction of the screening system of elite event before risk assessment process and quality control strategies for GM seed production. In addition, we suggested that the developers of GM crops should understand the importance of risk assessment and management for the commercialization of those crops and consider the biological and ecological characteristics of host plants. Consistent communications may need to be established between GM crop developers, risk assessors and risk managers at the initial stages of GM crop development to reduce trial-and-errors.

Genetics of Fusarium Wilt Resistance in Pigeonpea (Cajanus cajan) and Efficacy of Associated SSR Markers

  • Singh, Deepu;Sinha, B.;Rai, V.P.;Singh, M.N.;Singh, D.K.;Kumar, R.;Singh, A.K.
    • The Plant Pathology Journal
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    • 제32권2호
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    • pp.95-101
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    • 2016
  • Inheritance of resistance to Fusarium wilt (FW) disease caused by Fusarium udum was investigated in pigeonpea using four different long duration FW resistant genotypes viz., BDN-2004-1, BDN-2001-9, BWR-133 and IPA-234. Based on the $F_2$ segregation pattern, FW resistance has been reported to be governed by one dominant gene in BDN-2004-1 and BDN-2001-9, two duplicate dominant genes in BWR-133 and two dominant complimentary genes in resistance source IPA-234. Further, the efficacy of six simple sequence repeat (SSR) markers namely, ASSR-1, ASSR-23, ASSR-148, ASSR-229, ASSR-363 and ASSR-366 reported to be associated with FW resistance were also tested and concluded that markers ASSR-1, ASSR-23, ASSR-148 will be used for screening of parental genotypes in pigeonpea FW resistance breeding programs. The information on genetics of FW resistance generated from this study would be used, to introgress FW resistance into susceptible but highly adopted cultivars through marker-assisted backcross breeding and in conventional breeding programs.

Cloning and Characterization of a Novel ${\alpha}$-Amylase from a Fecal Microbial Metagenome

  • Xu, Bo;Yang, Fuya;Xiong, Caiyun;Li, Junjun;Tang, Xianghua;Zhou, Junpei;Xie, Zhenrong;Ding, Junmei;Yang, Yunjuan;Huang, Zunxi
    • Journal of Microbiology and Biotechnology
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    • 제24권4호
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    • pp.447-452
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    • 2014
  • To isolate novel and useful microbial enzymes from uncultured gastrointestinal microorganisms, a fecal microbial metagenomic library of the pygmy loris was constructed. The library was screened for amylolytic activity, and 8 of 50,000 recombinant clones showed amylolytic activity. Subcloning and sequence analysis of a positive clone led to the identification a novel gene (amyPL) coding for ${\alpha}$-amylase. AmyPL was expressed in Escherichia coli BL21 (DE3) and the purified AmyPL was enzymatically characterized. This study is the first to report the molecular and biochemical characterization of a novel ${\alpha}$-amylase from a gastrointestinal metagenomic library.

3D Structure of Bacillus halodurans O-Methyltransferase, a Novel Bacterial O-Methyltransferase by Comparative Homology Modeling

  • Lee, Jee-Young;Lee, Sung-Ah;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • 제28권6호
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    • pp.941-946
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    • 2007
  • Bacillus halodurans O-methyltransferase (BhOMT) is a S-adenosylmethionine (SAM or AdoMet) dependent methyltransferase. Three dimensional structure of the BhOMT bound to S-adenosyl-L-homocysteine (SAH or AdoHcy) has been determined by comparative homology modeling. BhOMT has 40% sequence identity with caffeoyl-CoA 3-O-methyltransferase (CCoAOMT) from alfalfa. Based on x-ray structure of CCoAOMT, three dimensional structure of BhOMT was determined using MODELLER. The substrate binding sites of these two proteins showed slight differences, but these differences were important to characterize the substrate of BhOMT. Automated docking study showed that four flavonoids, quercetin, fisetin, myricetin, and luteolin which have two hydroxyl groups simultaneously at 3'- and 4'-position in the B-ring and structural rigidity of Cring resulting from the double bond characters between C2 and C3, were well docked as ligands of BhOMT. These flavonoids form stable hydrogen bondings with K211, R170, and hydroxyl group at 3'-position in the Bring has stable electrostatic interaction with Ca2+ ion in BhOMT. This study will be helpful to understand the biochemical function of BhOMT as an O-methyltransferase for flavonoids.

인삼의 적변을 유발하는 세균에 대하여 항균활성을 가지는 방선균 선발 및 동정 (Screening and Identification of Antibacterial Actinomycetes against Bacteria Causing Rusty Root on Ginseng)

  • 한성희;류동걸;최승현;최재을;안길환
    • 농업과학연구
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    • 제37권2호
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    • pp.255-260
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    • 2010
  • Rusty root, the browning disease on ginseng, decreases quality and value. Recent studies indicated that endophytic bacteria could be a possible cause of rusty root. Actinomycetes antagonistic to the rusty-root-causing bacteria were isolated from soil. Twenty nine out of 932-isolates of Actinomycetes from soil showed antibacterial activity against Agrobacterium tumefaciens and Pseudomonas veronii an endophytic isolate in ginseng. The strongest antibacterial strain(ATO4O104) was classified based on 16S rDNA sequence. The Actinomycetes strain, ATO4O104, isolated in soil of USA volcano national park was identified as Streptomyces adephospholyticus. To test plant toxicity, radish seeds were sprouted with the culture of S. adephospholyticus and it did not show any harmful effect. The butanol partition out of n-hexane, ethyl acetate, butanol, and water partions showed the highest antibacterial activity.