• 제목/요약/키워드: Extracellular products

검색결과 201건 처리시간 0.023초

Purification and Characterization of a Thermostable Cellobiohydrolase from Fomitopsis pinicola

  • Shin, Keum;Kim, Yoon-Hee;Jeya, Marimuthu;Lee, Jung-Kul;Kim, Yeong-Suk
    • Journal of Microbiology and Biotechnology
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    • 제20권12호
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    • pp.1681-1688
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    • 2010
  • A screening for cellobiohydrolase (CBH) activity was performed and Fomitopsis pinicola KMJ812 was selected for further characterization as it produced a high level of CBH activity. An extracellular CBH was purified to homogeneity by sequential chromatography of F. pinicola culture supernatants. The molecular mass of the F. pinicola CBH was determined to be 64 kDa by SDS-PAGE and by size-exclusion chromatography, indicating that the enzyme is a monomer. The F. pinicola CBH showed a $t_{1/2}$ value of 42 h at $70^{\circ}C$ and catalytic efficiency of $15.8mM^{-1}s^{-1}(k_{cat}/K_m)$ for p-nitrophenyl-${\beta}$-D-cellobioside, one of the highest levels seen for CBH-producing microorganisms. Its internal amino acid sequences showed a significant homology with hydrolases from glycoside hydrolase family 7. Although CBHs have been purified and characterized from other sources, the F. pinicola CBH is distinguished from other CBHs by its high catalytic efficiency and thermostability.

Biotransformation of (-)-α-Pinene by Whole Cells of White Rot Fungi, Ceriporia sp. ZLY-2010 and Stereum hirsutum

  • Lee, Su-Yeon;Kim, Seon-Hong;Hong, Chang-Young;Kim, Ho-Young;Ryu, Sun-Hwa;Choi, In-Gyu
    • Mycobiology
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    • 제43권3호
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    • pp.297-302
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    • 2015
  • Two white rot fungi, Ceriporia sp. ZLY-2010 (CER) and Stereum hirsutum (STH) were used as biocatalysts for the biotransformation of (-)-${\alpha}$-pinene. After 96 hr, CER converted the bicyclic monoterpene hydrocarbon (-)-${\alpha}$-pinene into ${\alpha}$-terpineol (yield, 0.05 g/L), a monocyclic monoterpene alcohol, in addition to, other minor products. Using STH, verbenone was identified as the major biotransformed product, and minor products were myrtenol, camphor, and isopinocarveol. We did not observe any inhibitory effects of substrate or transformed products on mycelial growth of the fungi. The activities of fungal manganese-dependent peroxidase and laccase were monitored for 15 days to determine the enzymatic pathways related to the biotransformation of (-)-${\alpha}$-pinene. We concluded that a complex of enzymes, including intra- and extracellular enzymes, were involved in terpenoid biotransformation by white rot fungi.

국내산 침엽수 목분의 진탕배양에서 나타난 Fomitopsis palustris의 효소 활성 및 셀룰로오스 분해 (Enzyme Activities and Cellulose Degradation of Domestic Softwoods in Shaking Culture of Fomitopsis palustris)

  • 최두열;이영민;김영균;윤정준;김영숙
    • Journal of the Korean Wood Science and Technology
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    • 제35권6호
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    • pp.91-99
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    • 2007
  • 소나무, 낙엽송, 리기다소나무, 잣나무 등 국내산 침엽수 목분을 기질로 진탕배양했을 때 갈색부후균인 Fomitopsis palustris로부터 분비된 당 분해성 균체 외 효소는 Endoglucanase (EG), $\beta$-glucosidase (BGL) 및 $\beta$-1,4-xylosidase (BXL)와 함께 결정형 셀룰로오스를 분해하는 Cellobiohydrolase (CBH)도 활성을 갖은 것으로 나타났다. 4주간 배양에서 변재는 심재에 비해 큰 효소활성을 나타냈으나, 배양기간을 증가시킴으로써 효소활성이 커지는 것으로 밝혀져 효소에 의한 목질바이오매스 분해의 경우 심변재 혼합처리도 가능할 수 있음이 시사되었다. 그리고, 균체 의 단백질을 SDS-PAGE로 분석한 결과, 대부분의 수종에서 나타나는 효소의 단백질 패턴은 거의 유사한 것으로 나타나 효소를 이용한 목질계 바이오매스 분해의 경구 수종별 혼합처리도 가능함이 시사되었다. F. palustris에 의해 분해된 목분(60 mesh 통과) 셀룰로오스의 결정화도 감소율은 4주 배양에서 약 4.2~20.4%, 8주 배양에서 약 12.9~28.9% 수준으로 나타났다.

Arctigenin from Burdock Root Exhibits Potent Antibacterial and Anti-Virulence Properties against Pseudomonas aeruginosa

  • Abdulrahman E. Koshak;Mahmoud A. Elfaky;Hossam M. Abdallah;Dina A. I. Albadawi;Gamal A. Mohamed;Sabrin R. M. Ibrahim;Abdulrahim A. Alzain;El-Sayed Khafagy;Azza A. H. Rajab;Wael A. H. Hegazy
    • Journal of Microbiology and Biotechnology
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    • 제34권8호
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    • pp.1642-1652
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    • 2024
  • Arctium lappa (Burdock) root is used in various culinary applications especially in Asian Cuisine. Arctigenin (ARC) is a polyphenolic compound abundant in the roots of the burdock plant from which it derives its name. The emergence of bacterial resistance is a growing global worry, specifically due to the declining availability of new antibiotics. Screening for the antibacterial candidates among the safe natural products is a promising approach. The present study was aimed to assess the antibacterial activity of ARC against Pseudomonas aeruginosa exploring its effect on the bacterial cell membrane. Furthermore, the anti-virulence activities and anti-quorum sensing (QS) activities of ARC were in vitro, in vivo and in silico assessed against P. aeruginosa. The current results showed the ARC antibacterial activity was owed to its disruption effect of the cell membrane. ARC at sub-MIC significantly decreased the formation of biofilm, motility, production of extracellular enzymes and in vivo protected mice against P. aeruginosa. These anti-virulence activities of ARC are owed to its interference with bacterial QS and its expression. Furthermore, ARC showed mild effect on mammalian erythrocytes, low probability to induce resistance and synergistically combined with antibiotics. In summary, the promising anti-virulence properties of ARC indicate its potential as an effective supplement to conventional antibiotics for treating severe P. aeruginosa infections.

Microcystis sp. Cell의 부패와 염소 소독부산물 생성 (Decomposition of Microcystis sp. Cell and Formation of Chlorination Disinfection By-Products)

  • 손희종;염훈식;정종문;최진택
    • 대한환경공학회지
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    • 제34권5호
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    • pp.351-358
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    • 2012
  • Microcystis sp.의 부패과정에서 수중으로 용출되는 AOM 특성과 염소 이들에서의 disinfection by-products (DBPs) 생성특성을 조사하였다. 수중으로 용출된 EOM과 cell + IOM에서의 염소 DBPs 생성특성을 조사한 결과, EOM은 보관기간 초기부터 지속적으로 증가하였으나 cell + IOM의 경우는 급격한 감소경향을 나타내었으며, 생성된 DBPs 중 HAAFP가 가장 높은 생성비율을 나타내었다. 또한, 이 때의 DBP 구성종들의 변화를 살펴본 결과, HAA 구성종들의 경우는 EOM에서는 di-HAA 구성종들의 비율은 점점 감소하였고 tri-HAA 구성종들의 비율은 점점 증가하였다. 그러나 cell + IOM의 경우는 EOM의 경우와는 반대 경향을 나타내었다. 또한, HAN 구성종들의 경우는 EOM과 cell + IOM 모두 di-HAN 구성종들의 생성비율이 월등히 높았다.

Searching for Novel Candidate Small Molecules for Ameliorating Idiopathic Pulmonary Fibrosis: a Narrative Review

  • Kyung-il Kim;Rajib Hossain;Xin Li;Hyun Jae Lee;Choong Jae Lee
    • Biomolecules & Therapeutics
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    • 제31권5호
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    • pp.484-495
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    • 2023
  • Idiopathic pulmonary fibrosis (IPF) can be defined as a progressive chronic pulmonary disease showing scarring in the lung parenchyma, thereby resulting in increase in mortality and decrease in the quality of life. The pathophysiologic mechanism of fibrosis in IPF is still unclear. Repetitive microinjuries to alveolar epithelium with genetical predisposition and an abnormal restorative reaction accompanied by excessive deposition of collagens are involved in the pathogenesis. Although the two FDA-approved drugs, pirfenidone and nintedanib, are under use for retarding the decline in lung function of patients suffered from IPF, they are not able to improve the survival rate or quality of life. Therefore, a novel therapeutic agent acting on the major steps of the pathogenesis of disease and/or, at least, managing the clinical symptoms of IPF should be developed for the effective regulation of this incurable disease. In the present review, we tried to find a potential of managing the clinical symptoms of IPF by natural products derived from medicinal plants used for controlling the pulmonary inflammatory diseases in traditional Asian medicine. A multitude of natural products have been reported to exert an antifibrotic effect in vitro and in vivo through acting on the epithelial-mesenchymal transition pathway, transforming growth factor (TGF)- β-induced intracellular signaling, and the deposition of extracellular matrix. However, clinical antifibrotic efficacy of these natural products on IPF have not been elucidated yet. Thus, those effects should be proven by further examinations including the randomized clinical trials, in order to develop the ideal and optimal candidate for the therapeutics of IPF.

Trichoderma koningii에서 분리한 $\beta$-glucosidase의 작용양상 (The Action Mode of $\beta$-glucosidase Purified from Trichoderma koningii)

  • 정춘수;최지영;이헌주;맹필재;한인섭;강사욱;하영칠
    • 미생물학회지
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    • 제36권1호
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    • pp.8-13
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    • 2000
  • Trichoderma koningii ATCC 26113에서 분비되는 효소인 $\beta$-glucosidase를 cellobiose, sophorose, laminaribiose 및 gentiobiose 등의 기질과 반응시킨 후 효소의 transglycosylation 반응 산물을 분석하였다. 각각의 기질로부터 생성된 이당체(dimer)들을 HPLC로 분리하고 $^(1)H$-NMR spectroscopy를 통하여 분석하였다. Cellobiose를 기질로 사용하여 효소와 반응시켰을 때 그 산물에는 laminaribiose, sophorose 및 gentiobiose가 포함되었음을 확인할 수 있었다. Laminaribiose, sophorose 및 gentiobiose를 기질로 사용하였을 경우에 효소는 transglycosylation 반응을 통하여 새로운 $\beta$-glycosidic 결합을 갖는 이당체들을 생성하였다. 효소반응에 의하여 누적되는 이당체의 양은 생성속도보다는 분해속도에 의하여 결정되는 것으로 나타났다.

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Purification and Characterization of a Thermostable Xylanase from Fomitopsis pinicola

  • Shin, Keum;Jeya, Marimuthu;Lee, Jung-Kul;Kim, Yeong-Suk
    • Journal of Microbiology and Biotechnology
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    • 제20권10호
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    • pp.1415-1423
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    • 2010
  • An extracellular xylanase was purified to homogeneity by sequential chromatography of Fomitopsis pinicola culture supernatants on a DEAE-Sepharose column, a gel filtration column, and then on a MonoQ column with fast protein liquid chromatography. The relative molecular mass of the F. pinicola xylanase was determined to be 58 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis and by size-exclusion chromatography, indicating that the enzyme is a monomer. The hydrolytic activity of the xylanase had a pH optimum of 4.5 and a temperature optimum of $70^{\circ}C$. The enzyme showed a $t_{1/2}$ value of 33 h at $70^{\circ}C$ and catalytic efficiency ($k_{cat}=77.4\;s^{-1}$, $k_{cat}/K_m$=22.7 mg/ml/s) for oatspelt xylan. Its internal amino acid sequences showed a significant homology with hydrolases from glycoside hydrolase (GH) family 10, indicating that the F. pinicola xylanase is a member of GH family 10.

Organic Solvent and pH Induced Alteration of Product Specificity of CGTase

  • Park, Kyo-Sun;Oh, Hyun-Mi;Choe, Hui-Woog;Park, Chung-Ung;Lee, Kang-Min
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제3권2호
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    • pp.78-81
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    • 1998
  • Cyclodextrin glucanotransferase [CGTase, E.C.2.4.1.19] is an extracellular enzyme, which catalyzes he formation of ${\alpha}$-, ${\beta}$-, ${\gamma}$- CDs from starch. Their proportions of formations depend on enzyme sources and reaction conditions. To understand what determines the product specificity of CGTases, we examined the alteration of product specificity of CGTase from Bacillus macerans by organic solvent sand pH. At acidic pH range less than pH 6 where the enzyme was unstable, the ratio of ${\alpha}$-/ ${\beta}$-CD production was increased 4 times more than that at neutral pH range. As we increased the concentration of 2-butanol, ${\alpha}$-/ ${\beta}$-CD ratio was proportionally increased but / ratio remained constant. The ${\alpha}$-/ ${\beta}$-CD ratio of products was increased in the reaction media which yielded low products.

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NF-${\kappa}B$ and CREB Are Involved in IL-8 Production of Human Neutrophils Induced by Trichomonas vaginalis-Derived Secretory Products

  • Nam, Young-Hee;Min, Deul-Le;Park, Soon-Jung;Kim, Kyeong-Ah;Lee, Young-Ah;Shin, Myeong-Heon
    • Parasites, Hosts and Diseases
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    • 제49권3호
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    • pp.291-294
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    • 2011
  • Trichomonas vaginalis is a flagellated lumen-dwelling extracellular protozoan parasite that causes human trichomoniasis via sexual intercourse. Human neutrophils play a crucial role in acute tissue inflammatory responses in T. vaginalis infection. In this study, we investigated the signaling mechanism of neutrophil responses when stimulated with T. vaginalis-derived secretory products (TvSP), which were collected from $1{\times}10^7$ live trichomonads. Incubation of human neutrophils isolated from peripheral blood with TvSP induced up-regulation of IL-8 protein secretion. In addition, stimulation with TvSP induced phosphorylation of NF-${\kappa}B$ and CREB in neutrophils. Moreover, TvSP-induced IL-8 production was also significantly inhibited by pretreatment of neutrophils with $i{\kappa}B$ inhibitor or CREB inhibitor. These results suggest that transcription factors NF-${\kappa}B$ and CREB are involved in IL-8 production in human neutrophils induced by stimulation with T. vaginalis infection.