• 제목/요약/키워드: Metagenome

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Soil Metagenome to Explore Soil Microbial Resources

  • Lee, Seon-Woo;Lim, He-Kyoung;Kim, Sung-Eun;Choi, Gyung-Ja;Kim, Jin-Cheol;Choi, Kwang-Yun
    • 한국미생물생명공학회:학술대회논문집
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    • 한국미생물생명공학회 2003년도 2003 Annual Meeting, BioExhibition and International Symposium
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    • pp.20-23
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    • 2003
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Searching for Useful Enzymes from Metagenome Library

  • Ryu, Sangryeol;Yoon, Ji-Ae;Kang, Seo-Won;Park, Sul-Hee
    • 한국미생물생명공학회:학술대회논문집
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    • 한국미생물생명공학회 2003년도 2003 Annual Meeting, BioExhibition and International Symposium
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    • pp.14-17
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    • 2003
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Metagenome Analysis of Protein Domain Collocation within Cellulase Genes of Goat Rumen Microbes

  • Lim, SooYeon;Seo, Jaehyun;Choi, Hyunbong;Yoon, Duhak;Nam, Jungrye;Kim, Heebal;Cho, Seoae;Chang, Jongsoo
    • Asian-Australasian Journal of Animal Sciences
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    • 제26권8호
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    • pp.1144-1151
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    • 2013
  • In this study, protein domains with cellulase activity in goat rumen microbes were investigated using metagenomic and bioinformatic analyses. After the complete genome of goat rumen microbes was obtained using a shotgun sequencing method, 217,892,109 pair reads were filtered, including only those with 70% identity, 100-bp matches, and thresholds below $E^{-10}$ using METAIDBA. These filtered contigs were assembled and annotated using blastN against the NCBI nucleotide database. As a result, a microbial community structure with 1431 species was analyzed, among which Prevotella ruminicola 23 bacteria and Butyrivibrio proteoclasticus B316 were the dominant groups. In parallel, 201 sequences related with cellulase activities (EC.3.2.1.4) were obtained through blast searches using the enzyme.dat file provided by the NCBI database. After translating the nucleotide sequence into a protein sequence using Interproscan, 28 protein domains with cellulase activity were identified using the HMMER package with threshold E values below $10^{-5}$. Cellulase activity protein domain profiling showed that the major protein domains such as lipase GDSL, cellulase, and Glyco hydro 10 were present in bacterial species with strong cellulase activities. Furthermore, correlation plots clearly displayed the strong positive correlation between some protein domain groups, which was indicative of microbial adaption in the goat rumen based on feeding habits. This is the first metagenomic analysis of cellulase activity protein domains using bioinformatics from the goat rumen.

A Novel Esterase from a Marine Metagenomic Library Exhibiting Salt Tolerance Ability

  • Fang, Zeming;Li, Jingjing;Wang, Quan;Fang, Wei;Peng, Hui;Zhang, Xuecheng;Xiao, Yazhong
    • Journal of Microbiology and Biotechnology
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    • 제24권6호
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    • pp.771-780
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    • 2014
  • A putative lipolytic enzyme gene, named as est9x, was obtained from a marine microbial metagenome of the South China Sea. Sequence analysis showed that Est9X shares lower than 27% sequence identities with the characterized lipolytic enzymes, but possesses a catalytic triad highly conserved in lipolytic enzymes of the ${\alpha}/{\beta}$ hydrolase superfamily. By phylogenetic tree construction, Est9X was grouped into a new lipase/esterase family. To understand Est9X protein in depth, it was recombinantly expressed, purified, and biochemically characterized. Within potential hydrolytic activities, only lipase/esterase activity was detected for Est9X, confirming its identity as a lipolytic enzyme. When using p-nitrophenol esters with varying lengths of fatty acid as substrates, Est9X exhibited the highest activity to the C2 substrate, indicating it is an esterase. The optimal activity of Est9X occurred at a temperature of $65^{\cric}C$, and Est9X was pretty stable below the optimum temperature. Distinguished from other salt-tolerant esterases, Est9X's activity was tolerant to and even promoted by as high as 4 M NaCl. Our results imply that Est9X is a unique esterase and could be a potential candidate for industrial application under extreme conditions.

Cloning and Characterization of a ${\beta}$-Glucosidase from Marine Microbial Metagenome with Excellent Glucose Tolerance

  • Fang, Zemin;Fang, Wei;Liu, Juanjuan;Hong, Yuzhi;Peng, Hui;Zhang, Xuecheng;Sun, Baolin;Xiao, Yazhong
    • Journal of Microbiology and Biotechnology
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    • 제20권9호
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    • pp.1351-1358
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    • 2010
  • The demand for ${\beta}$-glucosidases insensitive to product inhibition is increasing in modern biotechnology, for these enzymes would improve the process of saccharification of lignocellulosic materials. In this study, a ${\beta}$-glucosidase gene that encodes a 442-amino-acid protein was isolated from a marine microbial metagenomic library by functional screening and named as bgl1A. The protein was identified to be a member of the glycoside hydrolases 1 family, and was recombinantly expressed, purified, and biochemically characterized. The recombinant ${\beta}$-glucosidase, Bgl1A, exhibited a high level of stability in the presence of various cations and high concentrations of NaCl. Interestingly, it was activated by glucose at concentrations lower than 400 mM. With glucose further increasing, the enzyme activity of Bgl1A was gradually inhibited, but remained 50% of the original value in even as high as 1,000 mM glucose. These findings indicate that Bgl1A might be a potent candidate for industrial applications.

자연발효 과정에서 인디고에 환원력을 지닌 미생물 커뮤니티 분석과 농화배양 (Analysis and Enrichment of Microbial Community Showing Reducing Ability toward indigo in the Natural Fermentation of Indigo-Plant)

  • 최은실;이은빈;최형안;손경희;김근중;신윤숙
    • KSBB Journal
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    • 제28권5호
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    • pp.295-302
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    • 2013
  • Indigo is utilized in various industries including textile dyeing, cosmetics, printing and medicinal products and its reduced form, leuco-indigo, is mainly used in these process. Chemical reducing agent (sodium dithionite, sodium sulfide, etc.) is preferred to use for the formation of leucoindigo in industry. In traditional indigo fermentation process, microorganisms can participate in the reduction of indigo and thus it has been known to reduce environmental pollution and noxious byproducts. However, in fermentation method using microorganisms it is difficult to standardize large scale production process due to low yield and reproducibility. In this study, we attempted to develop the indigo reduction process using microbial flora which was isolated from naturally fermented indigo vat or deduced by metagenomic approach. From the results of library analyses of PCR-amplified 16S rRNA genes from the traditional indigo fermentation vat sample (metagenome), it was confirmed that Alkalibacteriums (71%) was distinctly dominant in population. Some strains were identified after confirming that they become pure culture in nutrient media modified slightly. Four strains were separated in this process and each strain showed obvious reducing ability toward indigo in dyeing test. It is expected that the analyzed results will provide important data for standardizing the natural fermentation of indigo and investigating the mechanism of indigo reduction.

고속 염기서열 분석법을 이용한 자동차 공조 시스템(HAVC systems)의 악취 연관 곰팡이 군집 특성 (Characterization of odor-associated fungal community in automobile HVAC systems using a high-throughput DNA sequencing method)

  • 이윤영;최형주;윤정희;류희욱;조종래;성광모;조경숙
    • 실내환경 및 냄새 학회지
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    • 제16권1호
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    • pp.54-63
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    • 2017
  • The Automobile HVAC system is a habitat for odor-associated fungal communities. We investigated the odor-associated fungal community in an automobile HVAC system using a high-throughput DNA sequencing method. The fungal community structure was evaluated via metagenome analysis. At the phylum level, Ascomycota and Basidiomycota were detected, accounting for 43.41% and 56.49% of the fungal community in the HVAC system, respectively. Columnosphaeria (8.31%), Didymella (5.60%), Davidiella (5.50%), Microxyphium (4.24%), unclassified Pleosporales (2.90%), and Cladosporium (2.79%) were abundant at phylum of Ascomycota and Christiansenia (36.72%), Rhodotorula (10.48%), and Sporidiobolus (2.34%) were abundant at phylum of Basidiomycota. A total of 22 genera of fungi were isolated and identified from the evaporators of the HVAC systems which support fungal growth and biofilm formation. Among them, Cladosporium, Penicillium, Aspergillus, and Alternaria are the most representative odor-associated fungi in HVAC systems. They were reported to form biofilm on the surface of HVAC systems with other bacteria by hypha. In addition, they produce various mVOCs such as 3-methyl-1-butanol, acetic acid, butanoic acid, and methyl isobutyl ketone. Our findings may be useful for extending the understanding of odor-associated fungal communities in automobile HVAC systems.

High-throughput sequencing-based metagenomic and transcriptomic analysis of intestine in piglets infected with salmonella

  • KyeongHye, Won;Dohyun, Kim;Donghyun, Shin;Jin, Hur;Hak-Kyo, Lee;Jaeyoung, Heo;Jae-Don, Oh
    • Journal of Animal Science and Technology
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    • 제64권6호
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    • pp.1144-1172
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    • 2022
  • Salmonella enterica serovar Typhimurium isolate HJL777 is a virulent bacterial strain in pigs. The high rate of salmonella infection are at high risk of non-typhoidal salmonella gastroenteritis development. Salmonellosis is most common in young pigs. We investigated changes in gut microbiota and biological function in piglets infected with salmonella via analysis of rectal fecal metagenome and intestinal transcriptome using 16S rRNA and RNA sequencing. We identified a decrease in Bacteroides and increase in harmful bacteria such as Spirochaetes and Proteobacteria by microbial community analysis. We predicted that reduction of Bacteroides by salmonella infection causes proliferation of salmonella and harmful bacteria that can cause an intestinal inflammatory response. Functional profiling of microbial communities in piglets with salmonella infection showed increasing lipid metabolism associated with proliferation of harmful bacteria and inflammatory responses. Transcriptome analysis identified 31 differentially expressed genes. Using gene ontology and Innate Immune Database analysis, we identified that BGN, DCN, ZFPM2 and BPI genes were involved in extracellular and immune mechanisms, specifically salmonella adhesion to host cells and inflammatory responses during infection. We confirmed alterations in gut microbiota and biological function during salmonella infection in piglets. Our findings will help prevent disease and improve productivity in the swine industry.