• Title/Summary/Keyword: gram-scale

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Evaluation of Leachate Replacement Effect on Solid State Anaerobic Digestion of Dairy Manure and Sawdust Bedding Mixtures

  • Jo, Hyeonsoo;Lee, Seunghun;Kim, Eunjong;Ahn, Heekwon
    • Korean Journal of Soil Science and Fertilizer
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    • v.50 no.4
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    • pp.293-305
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    • 2017
  • This experiment was conducted to evaluate the effect of leachate replacement frequency on solid state anaerobic digestion (SSAD) of dairy manure using 22 L volume lab-scale digesters at mesophilic temperature ($37^{\circ}C$) in batch mode. Three different leachate replacement strategies (no replacement, once every three days, and once every nine days) were applied and three digesters per each treatment were operated for 45 days. Results showed that leachate replacement test unit every nine days resulted in 1.6 times more methane production ($53.8N{\cdot}mL\;g^{-1}{\cdot}VS$) from SSAD compared to test unit every three days ($34.0N{\cdot}mL\;g^{-1}{\cdot}VS$). No leachate replacement strategy applied group showed slightly higher methane production ($56.3N{\cdot}mL\;g^{-1}{\cdot}VS$) than every nine days replaced one. When added the methane production potential of replaced leachate itself to the methane produced from digester, leachate replacement every nine days resulted in quite similar methane production ($56.5N{\cdot}mL\;g^{-1}{\cdot}VS$) to no leachate replacement group. Even though methane production potential of replaced leachate itself added to the methane produced from digester, every three days replacement showed only $34N{\cdot}mL$ methane production per gram of volatile solids. These results suggest that farmers do not need to replace leachate during SSAD of dairy manure and sawdust mixture in order to maximize methane production. If there are any concerns with accumulation of inhibiting substances in the digester, the 9-day cycle leachate replacement is appropriate.

Functional Red Pigment Production in Solid-state Fermentation of Barley by Monascus sp. EBE1. (보리를 이용한 Monascus sp. EBE1 고상발효에 의한 기능성 적색 색소 생산)

  • 조창현;서동진;우건조;강대경
    • Microbiology and Biotechnology Letters
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    • v.30 no.3
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    • pp.253-257
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    • 2002
  • The time-dependent changes of red pigments production in solid-state plant scale fermentor using barley cultured with Monascus sp., instead of rice which was traditionally used, were investigated in this study. A steady increase in the yield of red pigments in barley occurred between the 3rd and 6th days. The optimized conditions (inoculation volume = 6∼8%, initial pH = 6, air supply = 0.6∼0.8 m) promoted the production of red pigments. Short-time steaming of barley (< 20 min) decreased fungal growth and pigments production due to the insufficient gelatiniza-tion. The optical density of the red pigments under the optimized conditions was 120 at 500 ]nm per gram of barley. In addition, the metabolites from the fermented barley with Monucus sp. showed antibacterial effects against Escherichia coli and Salmonella typhimurium. Barley was shown to be one of the best grain sources for solid-state fermentation with Monascu sp., fur obtaining natural pigments and also functional food materials.

The Stability, and Efficacy Against Penicillin-Resistant Enterococcus faecium, of the Plectasin Peptide Efficiently Produced by Escherichia coli

  • Chen, Xin;Wen, Yaoan;Li, Ling;Shi, Jiawei;Zhu, Zhe;Luo, Yuwen;Li, Yun;Chen, Rui
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1007-1014
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    • 2015
  • Plectasin, the first defensin extracted from a fungus (the saprophytic ascomycete Pseudoplectania nigrella), is attractive as a prospective antimicrobial agent. The purpose of this study was to establish a bacterium-based production system and evaluate the antimicrobial activity of the resulting plectasin. A gene encoding plectasin, with the codon preference of Escherichia coli, was optimized based on its amino acid sequence, synthesized using genesplicing with overlap extension PCR, and inserted into the expression vector pGEX-4T-1. The fusion protein was expressed in the soluble fraction of E. coli and purified using glutathione Stransferase affinity chromatography. Plectasin was cleaved from the fusion protein with thrombin and purified by ultrafiltration. The purified plectasin showed strong, concentrationdependent antimicrobial activity against gram-positive bacteria, including antibiotic-resistant bacteria, especially penicillin-resistant Enterococcus faecium. This antimicrobial activity was equal to chemically synthesized plectasin and was maintained over a wide range of pH and temperatures. This soluble recombinant expression system in E. coli is effective for producing plectasin at a relatively lower cost, and higher purity and efficiency than prior systems, and might provide a foundation for developing a large-scale production system. Overall, plectasin shows potential as a novel, high-performance, and safe antibiotic for the treatment of refractory diseases caused by drug-resistant bacterial strains.

Taxol Production in Taxus sap. Cell Cultures 1. Studies on Taxol Content in Yew Trees and Cultured Plant Cells (주목 세포배양에 의한 Taxoll 생산 1.주목 부위 및 서식 고도별 Taxol 함량 및 세포주 유도에 관한 연구)

  • 강인선;전정욱
    • KSBB Journal
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    • v.9 no.3
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    • pp.299-305
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    • 1994
  • Taxol contents in various parts of 15 years old yew tree were determined. The descending order of taxol content per unit mass was stem bark, root bark, needle and seed. In the seed, that order was seed coat, embryo and endosperm. The total amount of taxol extractable from a 15 years old yew tree was 1.68 gram. This amount was distributed in needle, stem bark, root bark and seeds as 48.0, 23.8, 27.9 and 0.4%, respectively. Altitudinal variation of taxol content was also observed. More taxol was observed in yew trees grown at high altitude over 1000m above sea level. Calli and suspension cultures were induced from various yew trees. The presence of taxo] in cultured cells was established bv HPLC. The taxol content in cultured cells were different according to the source of explants. These results may be useful for the goal of large scale taxol production by cultured yew tree cells.

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Preparation of a Vibrio vulnificus Vaccine with Immunogenicity and Protective Efficacy

  • Lee, Na-Gyong;Jung, Sang-Bo;Ahn, Bo-Young;Kim, Young-Gi;Kim, Je-Hak;Lee, Youn-Ha;Park, Wan-Je;Kim, Hyun-Su
    • Journal of Microbiology and Biotechnology
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    • v.7 no.6
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    • pp.423-428
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    • 1997
  • Vibrio vulnificus is a halophilic gram-negative human pathogen, which affects people with underlying liver diseases or a suppressed immune system, often leading to primary septicemia with a mortality rate of higher than 60%. In an effort to develop an oral vaccine against V. vulnificus infection, we prepared a whole cell killed vaccine of V. vulnificus on a large scale and compared the immunogenicity and protective efficacy of the vaccine administered in three formulation forms in rabbits. Since V. vulnificus O-antigen serotypes 1, 2, 3, 4, 5, and 7 account for more than 95% of clinical isolates, we prepared cell lysates from these six serotype strains and mixed in equal amounts for a vaccine. The vaccine was administered to rabbits intramuscularly (i.m.), orally as granules or as enteric-coated granules. In rabbits, all three formulation forms elicited a high level of serum IgG antibody reactive not only to the six strains but also to other O-antigen serotypes 6, 8 and 9, indicating cross-reactivities among the strains. Immunotherapeutic efficacy of the antisera was also evaluated by a passive immunization assay, which revealed that the orally immunized antisera as well as the i.m. immunized antisera was protective against a subsequent lethal challenge of V. vulnificus. These data demonstrate that oral immunization with a V. vulnificus whole cell lysate vaccine induced a systemic immune response and suggest the feasibility of development of this vaccine preparation as an oral vaccine.

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Expression and Purification of Transmembrane Protein MerE from Mercury-Resistant Bacillus cereus

  • Amin, Aatif;Sarwar, Arslan;Saleem, Mushtaq A.;Latif, Zakia;Opella, Stanley J.
    • Journal of Microbiology and Biotechnology
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    • v.29 no.2
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    • pp.274-282
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    • 2019
  • Mercury-resistant ($Hg^R$) bacteria were isolated from heavy metal polluted wastewater and soil collected near to tanneries of district Kasur, Pakistan. Bacterial isolates AZ-1, AZ-2 and AZ-3 showed resistance up to $40{\mu}g/ml$ against mercuric chloride ($HgCl_2$). 16S rDNA ribotyping and phylogenetic analysis were performed for the characterization of selected isolates as Bacillus sp. AZ-1 (KT270477), Bacillus cereus AZ-2 (KT270478) and Bacillus cereus AZ-3 (KT270479). Phylogenetic relationship on the basis of merA nucleotide sequence confirmed 51-100% homology with the corresponding region of the merA gene of already reported mercury-resistant Gram-positive bacteria. The merE gene involved in the transportation of elemental mercury ($Hg^0$) via cell membrane was cloned for the first time into pHLV vector and transformed in overexpressed C43(DE3) E. coli cells. The recombinant plasmid (pHLMerE) was expressed and the native MerE protein was obtained after thrombin cleavage by size exclusion chromatography (SEC). The purification of fusion/recombinant and native protein MerE by Ni-NTA column, dialysis and fast protein liquid chromatography (FPLC/SEC) involved unfolding/refolding techniques. A small-scale reservoir of wastewater containing $30{\mu}g/ml$ of $HgCl_2$ was designed to check the detoxification ability of selected strains. It resulted in 83% detoxification of mercury by B. cereus AZ-2 and B. cereus AZ-3, and 76% detoxification by Bacillus sp. AZ-1 respectively (p < 0.05).

Crystal Structure of SAV0927 and Its Functional Implications

  • Jeong, Soyeon;Kim, Hyo Jung;Ha, Nam-Chul;Kwon, Ae-Ran
    • Journal of Microbiology and Biotechnology
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    • v.29 no.3
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    • pp.500-505
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    • 2019
  • Staphylococcus aureus is a round-shaped, gram-positive bacterium that can cause numerous infectious diseases ranging from mild infections such as skin infections and food poisoning to life-threatening infections such as sepsis, endocarditis and toxic shock syndrome. Various antibiotic-resistant strains of S. aureus have frequently emerged, threatening human lives significantly. Despite much research on the genetics of S. aureus, many of its genes remain unknown functionally and structurally. To counteract its toxins and to prevent the antibiotic resistance of S. aureus, our understanding of S. aureus should be increased at the proteomic scale. SAV0927 was first sequenced in an antibiotic resistant S. aureus strain. The gene is a conserved hypothetical protein, and its homologues appear to be restricted to Firmicutes. In this study, we determined the crystal structure of SAV0927 at $2.5{\AA}$ resolution. The protein was primarily dimeric both in solution and in the crystals. The asymmetric unit contained five dimers that are stacked linearly with ${\sim}80^{\circ}$ rotation by each dimer, and these interactions further continued in the crystal packing, resulting in a long linear polymer. The crystal structures, together with the network analysis, provide functional implications for the SAV0927-mediated protein network.

Low Impact Development Scenario Designing for Large-scale Urban Development using SWMM-ING Software (SWMM-ING 소프트웨어를 이용한 대규모 도시개발부지에 대한 저영향개발 시나리오 설계)

  • Woo, Won Hee;Lee, Hanyong;Choi, Hyun Seok;Park, Youn Shik
    • Proceedings of the Korea Water Resources Association Conference
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    • 2021.06a
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    • pp.304-304
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    • 2021
  • 도시화가 진행됨에 따라 불투수지역이 늘어나는 등 개발전과 다른 지표 및 지표하 유출이 나타나며, 이러한 변화는 생태적, 수문학적으로 대규모 변화가 나타나는 것을 의미한다. 이에 따라 물관리, 물순환 등 물 지속 가능성은 대중의 관심을 받아 법, 제도가 변화하며 도시화에 대한 세심한 접근과 관리에 대한 계획이 대두되고 있는 실정이다. 저영향개발(Low Impact Development)은 이러한 도시화로 인해 발생될 수 있는 문제를 해결하기 위한 방안의 하나로 제안되고 있으며 첨두 및 유출총량을 완화하여 개발 이전 상태의 수문학적 기능을 유지하거나 물리학 및 생물학적 기작을 통해 오염물질의 저감효과도 나타나 대규모 도시개발 및 유역관리 사업에서 저영향개발기법의 적용이 점차 늘어나고 있는 추세이다. 그러나 저영향개발기법 설치에 관한 구체적인 매뉴얼 또는 가이드라인이 부족한 실정이기 때문에 저영향개발기법이 계획단계에서 양적으로 무분별하게 적용되어 비용적 측면에서 시공자에게 부담을 가중시키는 등의 문제가 발생한다. 또한 시설의 위치에 따라 강우유출수 및 비점오염원 저감에 대한 효과가 상이하기 때문에 시설의 적정 위치선정 및 시설용량, 경제성을 고려한 시설효과 검토 등 저영향개발기법 적용시 사업부지 특성에 따른 최적화를 고려하여 계획 및 설계에 적용할 필요성이 있다. 이에 비용적 측면을 고려한 최적설계가 가능한 SWMM-ING(Storm Water Management Model - Low impact development desigN proGram)을 개발된 바 있다. SWMM-ING은 비전문가도 사용이 가능하도록 저영향개발기법 최적화과정이 간단하고 직관적이며 비용을 고려한 최적설계가 가능하도록 개발된 소프트웨어로 기법 적용시 개략적인 비용추정 및 배치선정 등 저영향개발기법의 설계최적화가 가능하다. 또한 각각의 저영향개발기법에 대한 초기설치비용, 유지관리비용, 이자율, 기대수명, 대상지역 내 공간적 위치를 반영하여 강우유출수 및 비점오염저감효과를 시나리오별로 제시가 가능한 장점이 있다. 본 연구에서는 SWMM-ING를 이용하여 대규모 도시개발부지의 저영향개발기법 적용에 따른 유량 및 수질저감효과를 평가하고 유지관리비를 포함한 비용을 추정하여 최적설계 검토를 수행하였다.

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Biotechnology for the Mitigation of Methane Emission from Landfills (매립지의 메탄 배출 저감을 위한 생물공학기술)

  • Cho, Kyung-Suk;Ryu, Hee-Wook
    • Microbiology and Biotechnology Letters
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    • v.37 no.4
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    • pp.293-305
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    • 2009
  • Methane, as a greenhouse gas, is some 21~25 times more detrimental to the environmental than carbon dioxide. Landfills generally constitute the most important anthropogenic source, and methane emission from landfill was estimated as 35~73 Tg per year. Biological approaches using biocover (open system) and biofilter (closed system) can be a promising solution for older and/or smaller landfills where the methane production is too low for energy recovery or flaring and installation of a gas extraction system is inefficient. Methanotrophic bacteria, utilizing methane as a sole carbon and energy source, are responsible for the aerobic degradation (oxidation) of methane in the biological systems. Many bench-scale studies have demonstrated a high oxidation capacity in diverse filter bed materials such as soil, compost, earthworm cast and etc. Compost had been most often employed in the biological systems, and the methane oxidation rates in compost biocovers/boifilters ranged from 50 to $700\;g-CH_4\;m^{-2}\;d^{-1}$. Some preliminary field trials have showed the suitability of biocovers/biofilters for practical application and their satisfactory performance in mitigation methane emissions. Since the reduction of landfill methane emissions has been linked to carbon credits and trading schemes, the verified quantification of mitigated emissions through biocovers/biofilters is very important. Therefore, the assessment of in situ biocovers/biofilters performance should be standardized, and the reliable quantification methods of methane reduction is necessary.

An Assessment of Environmental Characteristics Associated with the Level of Endotoxin Concentration in Hospital Lobbies (일부 종합 병원 로비의 공기 중 엔도톡신 농도에 미치는 환경 요인 평가)

  • Lee, Kyeong-Min;Yeom, Jeongkwan;Lee, Wonjae;Ryu, Seung-Hun;Park, Dongjin;Park, Dong-Uk
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.24 no.3
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    • pp.310-320
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    • 2014
  • Backgrounds: Endotoxin, which found in the outer membrane of the gram-negative bacteria cell wall, makes up almost all of the lipopolysaccharide(LPS). When people are exposed to endotoxin,it can result in diverse health effects such as an airway irritation and inflammation, fever, malaise, bronchitis, allergic asthma, toxic pneumonitis, hypersensitivity lung disease. Cases among the elderly, children or pregnant can occur more frequently than a healthy adult if they are repeatedly exposed to the existing endotoxin. Therefore, we investigated and assessed the environmental characteristics associated with the airborne endotoxin concentration level in six hospital lobbies. Method: Endotoxin from indoor air in six hospital lobbies was measured by an area sampling method and analyzed according to American Society for Testing and Materials International(ASTM international) E2144-01. Total suspended particulate(TSP), carbon dioxide($CO_2$), temperature and humidity were also measured by using direct reading measurements or airborne sampling equipment at the same time. Environmental characteristics were appropriately divided into two or three groups for a statistics analysis. One-way analysis variable(one-way ANOVA) was used to examine a difference of the endotoxin concentration, depending on the environmental characteristics. In addition, only variables with p-value(p<0.25) were eventually designed to the best model by using multiple regression analysis. Results: The correlation analysis result indicated that TSP(p=0.003) and $CO_2$(p<0.0001) levels were significantly associated with endotoxin concentration levels. In contrast, temperature(p<0.068) and humidity(p<0.365) were not associated with endotoxin concentration. Levels of endotoxin concentration were statistically different among the environmental characteristics of Service time(p=0.01), Establishment of hospital(p<0.001), Scale of hospital(p=0.01), Day average people using hospital(p=0.03), Cleaning time of lobby(p=0.05), Season(p<0.001), and Cleaning of ventilation system(p<0.001) according to ANOVA. Finally, the best model(Adjusted R-square=72%) that we designed through a multiple regression test included environmental characteristics related to Service time, Area of lobby, Season, Cleaning of ventilation system, and Temperature. Conclusions: According to this study, our result showed a normal level of endotoxin concentration in the hospital lobbies and found environmental management methods to reduce the level of endotoxin concentration to a minimum. Consequently, this study recognized to be requirement for the management of ventilation systems and an indoor temperature in order to reduce the level of endotoxin concentration in the hospital lobbies.