• 제목/요약/키워드: Microbiology Quality

검색결과 492건 처리시간 0.03초

Expression and Purification of a Functional Recombinant Aspartate Aminotransferase (AST) from Escherichia coli

  • Zou, Lihui;Zhao, Haijian;Wang, Daguang;Wang, Meng;Zhang, Chuanbao;Xiao, Fei
    • Journal of Microbiology and Biotechnology
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    • 제24권7호
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    • pp.998-1003
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    • 2014
  • Aspartate aminotransferase (AST; E.C. 2.6.1.1), a vitamin B6-dependent enzyme, preferentially promotes the mutual transformation of aspartate and ${\alpha}$-ketoglutarate to oxaloacetate and glutamate. It plays a key role in amino acid metabolism and has been widely recommended as a biomarker of liver and heart damage. Our study aimed to evaluate the extensive preparation of AST and its application in quality control in clinical laboratories. We describe a scheme to express and purify the 6His-AST fusion protein. An optimized sequence coding AST was synthesized and transformed into Escherichia coli BL21 (DE3) strain for protein expression. Ideally, the fusion protein has a volumetric productivity achieving 900 mg/l cultures. After affinity chromatography, the enzyme activity of purified AST reached 150,000 U/L. Commutability assessment between the engineered AST and standard AST from Roche suggested that the engineered AST was the better candidate for the reference material. Moreover, the AST showed high stability during long-term storage at $-20^{\circ}C$. In conclusion, the highly soluble 6His-tagged AST can become a convenient tool for supplying a much better and cheaper standard or reference material for the clinical laboratory.

Preparation and Improvement of Physicochemical and Functional Properties of Dietary Fiber from Corn Cob Fermented by Aspergillus niger

  • Yadi Zhou;Qijie Sun;Chao Teng;Mingchun Zhou;Guangsen Fan;Penghui Qu
    • Journal of Microbiology and Biotechnology
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    • 제34권2호
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    • pp.330-339
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    • 2024
  • Corn cobs were fermented with Aspergillus niger to produce soluble dietary fiber (SDF) of high quality and excellent food safety. In this work, the fermentation process was optimized by single-factor test and response surface methodology (RSM). The optimal fermentation conditions were determined to be a material-liquid ratio of 1:30, an inoculum concentration of 11%, a temperature of 32℃, a time of 6 days, and a shaking speed of 200 r/min. Under these conditions, the SDF yield of corn cob increased from 2.34% to 11.92%, and the ratio of soluble dietary fiber to total dietary fiber (SDF/TDF) reached 19.08%, meeting the requirements for high-quality dietary fiber (SDF/TDF of more than 10%). Scanning electron microscopy (SEM) and Fourier-transformed infrared spectroscopy (FT-IR) analysis revealed that the fermentation effectively degraded part of cellulose and hemicellulose, resulting in the formation of a loose and porous structure. After fermentation the water swelling capacity, water-holding capacity, and oil-holding capacity of the corn cob SDF were significantly improved and the adsorption capacity of glucose, cholesterol, and nitrite ions all increased by more than 20%. Moreover, the total phenolic content increased by 20.96%, which correlated with the higher antioxidant activity of SDF. Overall, the fermentation of corn cobs by A. niger increased the yield and enhanced the functional properties of dietary fiber (DF) as well.

Antimicrobial (BN/PE) Film Combined with Modified Atmosphere Packaging Extends the Shelf Life of Minimally Processed Fresh-Cut Iceberg Lettuce

  • Kang, Sun-Chul;Kim, Min-Jeong;Park, In-Sik;Choi, Ung-Kyu
    • Journal of Microbiology and Biotechnology
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    • 제18권3호
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    • pp.568-572
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    • 2008
  • This study was conducted to investigate the effect of modified atmosphere packaging (MAP) in combination with BN/PE film on the shelf life and quality of fresh-cut iceberg lettuce during cold storage. The total mesophilic population in the sample packed in BN/PE film under MAP conditions was dramatically reduced in comparison with that of PE film, PE film under MAP conditions, and BN/PE film. The $O_2$ concentration in the BN/PE film under MAP conditions decreased slightly as the storage period progressed. The coloration of the iceberg lettuce progressed the slowest when it was packaged in BN/PE film under MAP conditions, followed by BN/PE film, PE film, and PE film under MAP conditions. The shelf life of fresh-cut iceberg lettuce packaged in the BN/PE film under MAP conditions was extended by more than 2 days at $10^{\circ}C$ as compared with that of the BN/PE film in which the extension effect was more than 2 days longer than that of PE, PET, and OPP films.

Bacterial Community Structure Shifted by Geosmin in Granular Activated Carbon System of Water Treatment Plants

  • Pham, Ngoc Dung;Lee, Eun-Hee;Chae, Seon-Ha;Cho, Yongdeok;Shin, Hyejin;Son, Ahjeong
    • Journal of Microbiology and Biotechnology
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    • 제26권1호
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    • pp.99-109
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    • 2016
  • We investigated the relation between the presence of geosmin in water and the bacterial community structure within the granular activated carbon (GAC) system of water treatment plants in South Korea. GAC samples were collected in May and August of 2014 at three water treatment plants (Sungnam, Koyang, and Yeoncho in Korea). Dissolved organic carbon and geosmin were analyzed before and after GAC treatment. Geosmin was found in raw water from Sungnam and Koyang water treatment plants but not in that from Yeoncho water treatment plant. Interestingly, but not surprisingly, the 16S rRNA clone library indicated that the bacterial communities from the Sungnam and Koyang GAC systems were closely related to geosmin-degrading bacteria. Based on the phylogenetic tree and multidimensional scaling plot, bacterial clones from GAC under the influence of geosmin were clustered with Variovorax paradoxus strain DB 9b and Comamonas sp. DB mg. In other words, the presence of geosmin in water might have inevitably contributed to the growth of geosmin degraders within the respective GAC system.

Toward Complete Bacterial Genome Sequencing Through the Combined Use of Multiple Next-Generation Sequencing Platforms

  • Jeong, Haeyoung;Lee, Dae-Hee;Ryu, Choong-Min;Park, Seung-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제26권1호
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    • pp.207-212
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    • 2016
  • PacBio's long-read sequencing technologies can be successfully used for a complete bacterial genome assembly using recently developed non-hybrid assemblers in the absence of second-generation, high-quality short reads. However, standardized procedures that take into account multiple pre-existing second-generation sequencing platforms are scarce. In addition to Illumina HiSeq and Ion Torrent PGM-based genome sequencing results derived from previous studies, we generated further sequencing data, including from the PacBio RS II platform, and applied various bioinformatics tools to obtain complete genome assemblies for five bacterial strains. Our approach revealed that the hierarchical genome assembly process (HGAP) non-hybrid assembler resulted in nearly complete assemblies at a moderate coverage of ~75x, but that different versions produced non-compatible results requiring post processing. The other two platforms further improved the PacBio assembly through scaffolding and a final error correction.

Rhizopus Koji를 이용한 무증자 쌀탁주 양조 (Takju Brewing of Uncooked Rice Starch Using Rhizopus Koji)

  • 손순기;노영혼;배상면;김현진
    • 한국미생물·생명공학회지
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    • 제18권5호
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    • pp.506-510
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    • 1990
  • 쌀 전분을 이용하여 무증자 당화 발효법으로 쌀 탁주를 제조하였으며 증자 당화 발효법에 의한 종래적인 방법과 비교하였다. Rhizopus속 균주를 증자하지 않은 생쌀에 접종하여 통상적인 방법에 따라 koji를 제조하여 사용하였으며 원료미를 분쇄해여 증자하지 않은 상태로 발효하였다. 무증자 당화 발효의 경우 종래적 방법에 의한 발효의 경우보다는 최종 alcohol 함량이 pH 1.8 높았고 유기산 생성이 급격히 이루어져 별도의 pH 조정이 필요없었다. 효모의 숫자는 차이가 없었고 세균은 무증자의 경우 2단 사입 후 24시간 까지는 증자의 경우보다 많다가 발효가 종료될 시점에서는 비슷하였다. 발효가 끝난후 제성주로 하였을 때 무증자의 경우는 fusel oil 함량은 증자의 경우보다 적었고 아미노산 함량은 약 2배로 높았으며 원료미에서 유래된 생취(raw flavor)가 유지되었다.

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Optimization of Hydroxyl Radical Scavenging Activity of Exopolysaccharides from Inonotus obliquus in Submerged Fermentation Using Response Surface Methodology

  • Chen, Hui;Xu, Xiangqun;Zhu, Yang
    • Journal of Microbiology and Biotechnology
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    • 제20권4호
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    • pp.835-843
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    • 2010
  • The objectives of this study were to investigate the effect of fermentation medium on the hydroxyl radical scavenging activity of exopolysaccharides from Inonotus obliquus by response surface methodology (RSM). A two-level fractional factorial design was used to evaluate the effect of different components of the medium. Corn flour, peptone, and $KH_2PO_4$ were important factors significantly affecting hydroxyl radical scavenging activity. These selected variables were subsequently optimized using path of steepest ascent (descent), a central composite design, and response surface analysis. The optimal medium composition was (% w/v): corn flour 5.30, peptone 0.32, $KH_2PO_4$ 0.26, $MgSO_4$ 0.02, and $CaCl_2$ 0.01. Under the optimal condition, the hydroxyl radical scavenging rate (49.4%) was much higher than that using either basal fermentation medium (10.2%) and single variable optimization of fermentation medium (35.5%). The main monosaccharides components of the RSM optimized polysaccharides are rhamnose, arabinose, xylose, mannose, glucose, and galactose with molar proportion at 1.45%, 3.63%, 2.17%, 15.94%, 50.00%, and 26.81%.

간장양조용 원료대체에 관한 연구(제4보) 소맥글루텐의 이용 (Studies on the Substitution of Raw Material for Soy Sauce Part IV. Use of Wheat gluten)

  • 이제문;안순복;김유삼;홍윤명;유주현
    • 한국미생물·생명공학회지
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    • 제2권2호
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    • pp.89-93
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    • 1974
  • 간장의 원료중 콩을 전분공장에서 부산물로 나오는 소맥글루텐으로 대체하는데 있어 원료배합비에 따른 국중 효순 생산량의 변화를 검토하고 제국조건을 결정한 뒤 원료총량 6kg의 규모로 담금하여 숙성중 제 성분의 변화를 분석 검토하고 이의 관능검사를 하였다. 소맥글루텐으로 탈지대두의 60% 대체하므로써 원료 총량에대해 소맥글루텐의 함량이 30% 되게 이용하므로써 성분상으로나 관능면에서 기존 간장에 뒤지지 않는 양질의 간장을 얻을 수 있다. 이때 산수량은 100%, 제국기간은 3일국이 적당하다. 총 질소함량은 소맥글루텐의 증량과 더불어 증가하나 60% 대체(총원료의 30%) 했을 때 원료중 총질소 이용도가 가장 높은데, 이는 제국중 국중 효소생산량도 이때 가장 높았던 결과와 일치한다. 본 연구는 1971년도 과학기술처 조사연구용역계약에 의거 이루워졌으며 현재 이 결과의 일부를 공업용으로 이용되고있다. 이에 대하여 과학기술처에 사의를 표합니다.

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Saccharomyces와 Hansenula의 혼합배양에 의해 제조한 민속증류수의 향미특성 (Flavor Characteristics of Korean Traditional Distilled Liquors Produced by the Co-culture of Saccharomyces and Hansenula)

  • 홍연;박승국;최언호
    • 한국미생물·생명공학회지
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    • 제27권3호
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    • pp.236-245
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    • 1999
  • Andong Soju is a Korean traditional distilled liquor brewed with Nuruk which is cultured with wild microorganisms. To provide useful information for scientific production and systematic quality control of traditional distilled liquor, the effects of mixed culture of the alcoholic yeasts and saccharifying molds isolated from the Nuruk, and mashes on the flavor and sensory characteristics were investigated. Distillate from mashes cocultured with Saccharomyces cerevisiae and Hansenula anomala using Mucor Nuruk was compared with distillate from mashes brewed with Andong Nuruk and with distillate from plant fermented mashes to analyze their flavor characteristics. The volatile flavor compounds in distillates were analyzed by GC and GC-MS using direct injection, solvent extraction, and purge & trap methods. Alcohols such as 3-methyl-1-butanol, 2-methyl-1-propanol, 1-propanol, and 2-phenyl ethanol; aldehydes such as acetaldehyde and 2-furancar-boxaldehyde; esters such as ethyl ester of acetic acid, hexanoic acid, octanoic acid, decanoic acid; alkanes, alkenes, ketone, sulfur, and pyrone compounds were detected. Alcohols were chief components of flavor compounds. No significant difference in overall acceptability test was shown among three experimental groups(p<0.05), but Nuruk-like aroma, Kaoliangchiew-like aroma, sweet taste, and well rounded mouthfeel showed significant differences among them(p<0.05).

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Analysis of Expressed Sequence Tags from the Wood-Decaying Fungus Fomitopsis palustris and Identification of Potential Genes Involved in the Decay Process

  • Karim, Nurul;Shibuya, Hajime;Kikuchi, Taisei
    • Journal of Microbiology and Biotechnology
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    • 제21권4호
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    • pp.347-358
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    • 2011
  • Fomitopsis palustris, a brown-rot basidiomycete, causes the most destructive type of decay in wooden structures. In spite of its great economic importance, very little information is available at the molecular level regarding its complex decay process. To address this, we generated over 3,000 expressed sequence tags (ESTs) from a cDNA library constructed from F. palustris. Clustering of 3,095 high-quality ESTs resulted in a set of 1,403 putative unigenes comprising 485 contigs and 918 singlets. Homology searches based on BlastX analysis revealed that 78% of the F. palustris unigenes had a significant match to proteins deposited in the nonredundant databases. A subset of F. palustris unigenes showed similarity to the carbohydrateactive enzymes (CAZymes), including a range of glycosyl hydrolase (GH) family proteins. Some of these CAZyme-encoded genes were previously undescribed for F. palustris but predicted to have potential roles in biodegradation of wood. Among them, we identified and characterized a gene (FpCel45A) encoding the GH family 45 endoglucanase. Moreover, we also provided functional classification of 473 (34%) of F. palustris unigenes using the Gene Ontology hierarchy. The annotated EST data sets and related analysis may be useful in providing an initial insight into the genetic background of F. palustris.