• Title/Summary/Keyword: Metagenome

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Biochemical and Structural Analysis of Hormone-sensitive Lipase Homolog EstE7: Insight into the Stabilized Dimerization of HSL-Homolog Proteins

  • Nam, Ki-Hyun;Park, Sung-Ha;Lee, Won-Ho;Hwang, Kwang-Yeon
    • Bulletin of the Korean Chemical Society
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    • v.31 no.9
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    • pp.2627-2632
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    • 2010
  • Hormone sensitive lipase (HSL) plays a major role in energy homeostasis and lipid metabolism. Several crystal structures of HSL-homolog proteins have been identified, which has led to a better understanding of its molecular function. HSL-homolog proteins exit as both monomer and dimer, but the biochemical and structural basis for such oligomeric states has not been successfully elucidated. Therefore, we determined the crystal structure of HSL-homolog protein EstE7 from a metagenome library at $2.2\;{\AA}$ resolution and characterized the oligomeric states of EstE7 both structurally and biochemically. EstE7 protein prefers the dimeric state in solution, which is supported by its higher enzymatic activity in the dimeric state. In the crystal form, EstE7 protein shows two-types of dimeric interface. Specifically, dimerization via the external ${beta}8$-strand occurred through tight association between two pseudosymmetric folds via salt bridges, hydrogen bonds and van der Waals interactions. This dimer formation was similar to that of other HSL-homolog protein structures such as AFEST, BEFA, and EstE1. We anticipate that our results will provide insight into the oligomeric state of HSL-homolog proteins.

Development of a Meta-Information System for Microbial Resources

  • Yu Jae-Woo;Chung Won-Hyong;Sohn Tae-Kwon;Park Yong-Ha;Kim Hong-Ik
    • Journal of Microbiology and Biotechnology
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    • v.16 no.2
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    • pp.178-183
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    • 2006
  • Microbes are one of the most important bioresources in bioindustry and provide high economic values. Although there are currently about 6,000 bacterial species with validly published names, microbiologists generally assume that the number may account for less than 1% of the bacterial species present on Earth. To discover the remaining species, studies of metagenomes, metabolomes, and proteomes related to microbes have recently been carried out in various fields. We have constructed an information system that integrates various data on microbial resources and manages bioinformation to support efficient research of microorganisms. We have designated this system 'Bio-Meta Information System (Bio-MIS).' Bio-MIS consists of an integrated microbial resource database, a microbial resource input system, an integrated microbial resource search engine, a microbial resource online distribution system, a portal service, and management via the Internet. In the future, this system is expected to be connected with various public databases. We plan to implement useful bioinformatics software for analyzing microbial genome resources. The Web site is accessible at http://biomis.probionic.com.

Identification of Novel Phytase Genes from an Agricultural Soil-Derived Metagenome

  • Tan, Hao;Mooij, Marlies J.;Barret, Matthieu;Hegarty, Pardraig M.;Harrington, Catriona;Dobson, Alan D.W.;O'Gara, Fergal
    • Journal of Microbiology and Biotechnology
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    • v.24 no.1
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    • pp.113-118
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    • 2014
  • Environmental microorganisms are emerging as an important source of new enzymes for wide-scale industrial application. In this study, novel phytase genes were identified from a soil microbial community. For this, a function-based screening approach was utilized for the identification of phytase activity in a metagenomic library derived from an agricultural soil. Two novel phytases were identified. Interestingly, one of these phytases is an unusual histidine acid phosphatase family phytase, as the conserved motif of the active site of PhyX possesses an additional amino acid residue. The second phytase belongs to a new type, which is encoded by multiple open reading frames (ORFs) and is different to all phytases known to date, which are encoded by a single ORF.

Oral and Human Microbiome Research

  • Chung, Sung-Kyun
    • Journal of dental hygiene science
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    • v.19 no.2
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    • pp.77-85
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    • 2019
  • In the past gut microbiome has been the main focus of microbiome research. Studies about the microbiome inside oral cavities and other organs are underway. Studies about the relationship between noninfectious diseases and periodontal diseases, and the negative effects of harmful oral microbes on systemic health have been published in the recent past. A lot of attention is being paid towards fostering a healthy oral microbial ecosystem. This study aimed to understand the roles and effects of the microbiome inside the human body can potentially help cure various diseases including inflammatory bowel diseases with no known cure such as Crohn's disease, atopic dermatitis, obesity, cancer, diabetes, brain diseases and oral diseases. The present study examined technological trends in the correlation between the human microbiome and diseases in the human body, interactions between the human body's immunity, the metabolic system, and the microbiome, and research trends in other countries. While it has been proven that human microbiome is closely correlated with human diseases, most studies are still in the early stage of trying to compare the composition of microbiomes between health and patient groups. Since the oral environment is a dynamic environment that changes due to not only food intake but also other external factors such as lifestyle, hygiene, and drug intake, it is necessary to continue in-depth research on the microbiome composition characteristics to understand the complex functions of oral microorganisms. Analyzing the oral microbiome using computational technology may aid in disease diagnosis and prevention.

Oral Metagenomic Analysis Techniques

  • Chung, Sung-Kyun
    • Journal of dental hygiene science
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    • v.19 no.2
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    • pp.86-95
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    • 2019
  • The modern era of microbial genome analysis began in earnest in the 2000s with the generalization of metagenomics and gene sequencing techniques. Studying complex microbial community such as oral cavity and colon by a pure culture is considerably ineffective in terms of cost and time. Therefore, various techniques for genomic analysis have been developed to overcome the limitation of the culture method and to explore microbial communities existing in the natural environment at the gene level. Among these, DNA fingerprinting analysis and microarray chip have been used extensively; however, the most recent method of analysis is metagenomics. The study summarily examined the overview of metagenomics analysis techniques, as well as domestic and foreign studies on disease genomics and cluster analysis related to oral metagenome. The composition of oral bacteria also varies across different individuals, and it would become possible to analyze what change occurs in the human body depending on the activity of bacteria living in the oral cavity and what causality it has with diseases. Identification, isolation, metabolism, and presence of functional genes of microorganisms are being identified for correlation analysis based on oral microbial genome sequencing. For precise diagnosis and treatment of diseases based on microbiome, greater effort is needed for finding not only the causative microorganisms, but also indicators at gene level. Up to now, oral microbial studies have mostly involved metagenomics, but if metatranscriptomic, metaproteomic, and metabolomic approaches can be taken together for assessment of microbial genes and proteins that are expressed under specific conditions, then doing so can be more helpful for gaining comprehensive understanding.

Metagenomic Analysis of the Fecal Microbiomes of Wild Asian Elephants Reveals Microflora and Enzymes that Mainly Digest Hemicellulose

  • Zhang, Chengbo;Xu, Bo;Lu, Tao;Huang, Zunxi
    • Journal of Microbiology and Biotechnology
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    • v.29 no.8
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    • pp.1255-1265
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    • 2019
  • To investigate the diversity of gastrointestinal microflora and lignocellulose-degrading enzymes in wild Asian elephants, three of these animals living in the same group were selected for study from the Wild Elephant Valley in the Xishuangbanna Nature Reserve of Yunnan Province, China. Fresh fecal samples from the three wild Asian elephants were analyzed by metagenomic sequencing to study the diversity of their gastrointestinal microbes and cellulolytic enzymes. There were a high abundance of Firmicutes and a higher abundance of hemicellulose-degrading hydrolases than cellulose-degrading hydrolases in the wild Asian elephants. Furthermore, there were a high abundance and a rich diversity of carbohydrate active enzymes (CAZymes) obtained from the gene set annotation of the three samples, with the majority of them showing low identity with the CAZy database entry. About half of the CAZymes had no species source at the phylum or genus level. These indicated that the wild Asian elephants might possess greater ability to digest hemicellulose than cellulose to provide energy, and moreover, the gastrointestinal tracts of these pachyderms might be a potential source of novel efficient lignocellulose-degrading enzymes. Therefore, the exploitation and utilization of these enzyme resources could help us to alleviate the current energy crisis and ensure food security.

The Effect of Vacuum Films on Physicochemical and Microbiological Characteristics of Hanwoo (Korean Native Cattle)

  • Hwan Hee Yu;Sung Hun Yi;Sang-Dong Lim;Sang-Pil Hong
    • Food Science of Animal Resources
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    • v.43 no.3
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    • pp.441-453
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    • 2023
  • The objective of this study was to investigate the physicochemical and microbiological characteristics of round of Hanwoo by vacuum packaging film materials, polyvinylidene chloride (PVDC) and ethylene vinyl alcohol (EVOH). The packaged beef samples were stored in refrigerated conditions (2±1℃) for 12 weeks. Physicochemical analysis with pH, surface color, thiobarbituric acid reactive substances (TBARS) values, and volatile basic nitrogen (VBN) values and microbiological analysis with aerobic plate count (APC) and metagenomic analysis of packaged beef samples were performed. The pH and surface color did not change substantially during the 12 weeks and EVOH-packaged beef tended to be lower than those of PVDC-packaged beef. PVDC- and EVOH-packaged samples showed low TBARS and VBN values below standard limits. APC did not exceed 7 Log CFU/g for both samples during storage. In metagenomic analysis, Firmicutes and Lactobacillaceae were dominant phylum and family of the PVDC- and EVOH-packaged beef. In both packaged samples, Dellaglioa algida was the dominant species during storage, with the notable difference being the presence of Lactococcus piscium. Therefore, this study provided the information on the quality of vacuum-packaged beef according to different vacuum films for long-term refrigerated storage.

Antimicrobial active clones from soil metagenomic library

  • H. K. Lim;Lee, E. H;Kim, J.C.;Park, G. J.;K S. Jang;Park, Y. H.;K Y. Cho;S, W. Lee
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.108.1-108
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    • 2003
  • Soil metagenome is untapped total microbial genome including that of the majority of unculturable bacteria present in soil. We constructed soil metagenomic library in Escherichia coli using DNA directly extracted from two different soils, pine tree rhizosphere soil and forest topsoil. Metagenomic libraries constructed from pine tree rhizosphere soil and forest topsoil consisted of approximately 33,700 clones and 112,000 clones with average insert DNA size of 35-kb, respectively. Subsequently, we screened the libraries to select clones with antimicrobial activities against Saccharomyces cerevisiae and Agrobacterium tumefaciens using double agar layer method. So far, we have a clone active against S. cerevisiae and a clone active against A. tumefaciens from the forest topsoil library. In vitro mutagenesis and DNA sequence analysis of the antifungal clone revealed the genes involved in the biosynthesis of antimicrobial secondary metabolite. Metagenomic libraries constructed in this study would be subject to search for diverse genetic resources related with useful microbial products.

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In situ analysis of the bacterial community associated with the Korean salty fermented seafood jeotgal

  • Hyunjun Kim;Yoomin Ahn;Chulhee Park;Eungbin Kim
    • Korean Journal of Environmental Biology
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    • v.39 no.4
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    • pp.515-522
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    • 2021
  • Jeotgal is a salty and fermented traditional Korean fish sauce. Unlike most other previous studies that investigated samples purchased from retail markets, this study focused on samples of jeotgal with traceable history to Yeonggwang, a timehonored fishing village in Korea. Three jeotgal samples, which were made from small yellow croakers, largehead hairtail, and miscellaneous fish, were selected based on information obtained from interviews with local craftsmen and literature reviews. Bacterial community profiles of the three jeotgal samples were investigated to identify indicator (and potentially core) bacteria for jeotgal ripening. The 16S rRNA gene-based metagenomic analysis revealed that the dominant phyla and classes, (Gammaproteobacteria, Betaproteobacteria, Bacilli, and Clostridia) of the three different jeotgal were identical, albeit with different composition ratios. Diversification was evident beginning at the order level. Interestingly, each dominant order was mainly comprised of single members even at the genus level. The dominant genera included Halomonas, Tetragenococcus, Halanaerobium, Pseudomonas, Massilia, and Lentibacillus. This observed genus-level heterogeneity suggests that there are diverse bacterial signatures in jeotgal and that these can be used as indicators for jeotgal ripening and/or as starters to increase its sensory quality and functionality.

Comparative analysis of HiSeq3000 and BGISEQ-500 sequencing platform with shotgun metagenomic sequencing data

  • Animesh Kumar;Espen M. Robertsen;Nils P. Willassen;Juan Fu;Erik Hjerde
    • Genomics & Informatics
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    • v.21 no.4
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    • pp.49.1-49.11
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    • 2023
  • Recent advances in sequencing technologies and platforms have enabled to generate metagenomics sequences using different sequencing platforms. In this study, we analyzed and compared shotgun metagenomic sequences generated by HiSeq3000 and BGISEQ-500 platforms from 12 sediment samples collected across the Norwegian coast. Metagenomics DNA sequences were normalized to an equal number of bases for both platforms and further evaluated by using different taxonomic classifiers, reference databases, and assemblers. Normalized BGISEQ-500 sequences retained more reads and base counts after preprocessing, while a slightly higher fraction of HiSeq3000 sequences were taxonomically classified. Kaiju classified a higher percentage of reads relative to Kraken2 for both platforms, and comparison of reference database for taxonomic classification showed that MAR database outperformed RefSeq. Assembly using MEGAHIT produced longer assemblies and higher total contigs count in majority of HiSeq3000 samples than using metaSPAdes, but the assembly statistics notably improved with unprocessed or normalized reads. Our results indicate that both platforms perform comparably in terms of the percentage of taxonomically classified reads and assembled contig statistics for metagenomics samples. This study provides valuable insights for researchers in selecting an appropriate sequencing platform and bioinformatics pipeline for their metagenomics studies.