• 제목/요약/키워드: extracellular enzyme

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Bacillus licheniformis의 세포막 프로티아제 부분 정제 및 특성 (Extracellular proteases from bacillus licheniformis : partial purification and characterization)

  • 홍난숙;최명언;양철학
    • 미생물학회지
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    • 제27권3호
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    • pp.245-249
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    • 1989
  • Bacillus licheniforntis에서 분비된 단백질 추출물에서 ammonium sulfate fractionation과 Sephadex G-75젤 여과 크로마토그라피를 사용하여 부분 정제된 2개의 서로 다른 활성을 갖는 단백질 분해 효소를 얻었다. 단백질 분해 효소 type l은 카제인에 대해 비교적 높은 활성을 보이며 organotluoride와 EOTA에 의한 억제에 매우 민감했다 이 효소의 최적 pH는 7.5였으며 $75^{\circ}C$에서 빠르게 변성되었다. 단백질 분해효소 type II는 type I에 비해 낮은 카제인 분해 활성을 보이며 ImM PMSF와 10mM EOTA용액과 함께 $37^{\circ}C$에서 30분 간 반응시키면 역사 활성이 억제되었다 한편 이 효소 는 pH 8.0에서 최대 활성을 나타내고 $71^{\circ}C$ 에서 상대적으로 늦게 불환성화 되었다.

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Streptomyces속 115-5 균주로부터 생성된 Chitinase의 저해작용기작 (Some Kinetic Properties of an Extracellular Chitinase from Streptomyces sp, 115-5)

  • Hong, Yong-Ki;Seu, Jung-Hwn
    • 한국미생물·생명공학회지
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    • 제9권4호
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    • pp.179-183
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    • 1981
  • 진균류의 세포벽제거 및 이에 따른 protoplast생성 등에 많이 이용되어지고 있는 chitinase를 Streptomyces sp. 115-5 균주로부터 생산하여 순수 정제한 다음, 이 chitinase의 작용을 저해하는 포도당의 저해양상을 조사하였다. 포도당 외에 D-glucuronic acid, D-sorbitol및 D-xylose등도 chitinase의 활성을 저해하였다. 그런고로, 포도당 분자에 의한chitinase의 활성 저해 효과에는 위의 분자들의 공통부분인 2번, 3번 및 4번 탄소의 hydroxyl group들의 구조위치가 중요한 영향을 가진다는 것을 알 수 있다. 그리고 포도당에 의한 chitinase의 저해양상은 competitive inhibition 과 non-competitive inhibition 과의 혼합 저해형으로 나타났다.

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균체외 \alpha-galactosidase를 생산하는 Streptomyces sp. YB-4의 분리 및 효소 특성 (Characterization of Extracellular \alpha-galactosidase Produced by Streptomyces sp. YB-4.)

  • 김소영;조기행;김창진;박동진;윤기홍
    • 한국미생물·생명공학회지
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    • 제30권4호
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    • pp.332-338
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    • 2002
  • 토양으로부터 세포외로 $\alpha$-galactosidase를 분비 생산하는 방선균 YB-4가 분리되었으며, 분리균의 배양ㆍ형태ㆍ생리적 특성을 조사한 결과 Streptomyces 속 균주로 확인되었다. 분리균의 배양상등액으로부터 부분정제된 $\alpha$-galactosidase를 조효소액으로 사용하였을 때 para-nitrophenyl-$\alpha$-D-galacto-pyranoside는 pH 6.0과 6$0^{\circ}C$의 반응조건에서 가장 잘 분해되었으며, 조효소액을 pH 4.0에서 pH 10.0범위에서 1시간 이상 방치한 후에도 약 90% 이상의 $\alpha$-galactosidase 활성을 유지하였다. 또한 분리균이 생산하는 $\alpha$-galactosidase는 melibiose, raffinose와 stachyose와 같은 저당류를 가수분해 할 수 있으며 분해산물로 galactose를 방출하는 것으로 보아 $\alpha$-1,6 결합을 분해한 것으로 확인되었다.

Identification and Characterization of Fungi Contaminated in the Built-In Furniture of an Apartment Home

  • Choi, Min Ah;Ahn, Geum Ran;Kim, Seong Hwan
    • Mycobiology
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    • 제47권4호
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    • pp.430-440
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    • 2019
  • Fungal contamination of built-in furniture is a frequent problem in Korea when new apartment is built. However, domestic information on the contaminating fungi is very limited. This study was conducted to isolate, identify and characterize the fungi of the problem in one of the apartment houses where the fungi were claimed in the built-in furniture before the house owner moves in. Fungi present in the furniture installed in a main room, dress room, and kitchen side were visually and microscopically confirmed and purely isolated on PDA. The isolated fungi were identified by analyzing the morphological characteristics and nucleotide sequence of the ITS, calmodulin gene, and TEF-1α gene. Aspergillus creber, A. niger, A. pseudoglacus, A. ruber, Cladosporium perangustum and Penicillium commune were identified. Four out of the six fungal species were positive for at least one enzyme in six kinds of extracellular enzyme assays. When these four species (A. creber, A. niger, C. perangustum and P. commune) were inoculated onto four kinds of wood chips of furniture materials, they were able to colonize all of the wood chips. Their settlement was better at 95% humidity condition than at 30% humidity condition. Among the four species, C. perangustum caused the darkest discoloration and secreted the most number of extracellular enzymes. The four species were re-isolated from the colonized wood chips and confirmed as the problematic fungi in the built-in furniture.

호열성 사상균 Thermoascus aurantiacus의 알코올분해대사 관련 효소학적 특성 (Enzyme Production Related to Alcohol Metabolism from Thermophilic Fungus Thermoascus aurantiacus)

  • 고희선;김현수
    • 한국미생물·생명공학회지
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    • 제34권3호
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    • pp.216-220
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    • 2006
  • 본 균의 생육 및 효소생산에 유용한 탄소원으로서 자연계의 식물에 풍부한 펙틴을 탄소원으로 할 경우, 그 생육도는 전분보다 뛰어났으며, alcohol oxidase와 catalase의 생산량도 높아지는 것으로 나타났다. 특히 alcohol oxidase의 경우는 전분의 15배 이상의 생산량을 보여 본 균과 펙틴 이용성과의 관계를 시사하였고, 세포외 pectin esterase, pectinase등의 높은 활성이 검출되어 이를 증명하였다. 또한 alcohol oxidase 반응에서 생성되는 물질인 formaldehyde를 산화하는 formaldehyde dehydrogenase와, formate를 산화하여 $CO_2$를 생성하는 formate dehydrogenase의 반응을 발견하여, 본 균의 pectin 이용성과 관련한 일련의 에너지 대사계의 존재를 추정할 수 있었다.

Detection of Aspergillus, Penicillium, and Fusarium Species by Sandwich Enzyme-Linked Immunosorbent Assay Using Mixed Monoclonal Antibodies

  • Kwak, Bo-Yeon;Kwon, Byung-Joon;Kweon, Chang-Hee;Shon, Dong-Hwa
    • Journal of Microbiology and Biotechnology
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    • 제14권2호
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    • pp.385-389
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    • 2004
  • The antibody-mix sandwich enzyme-linked immunosorbent assay (Ab-mix sELlS A) system was developed in order to simultaneously detect the extracellular polysaccharide (FPS) of Aspergillus, Penicillium, or Fusarium species using one detection system. The detection limit and detection range of the Ab-mix sELISA towards EPS of Penicilliun citrinum were not changed, and those towards Fusarium moniliforme EPS were changed a little compared to that of individual sandwich ELISA [9, 10]. The fungal culture filtrates of Aspergillus and Penicillium species showed nearly similar reactivity towards Ab-mix sELISA as that of sELISA using the MAb lB8 alone [9]. Also, the fungal culture filtrates of Fusarium species showed nearly the same reactivity towards Ab-mix sELISA as that of sELISA using the MAb lB8 alone [10]. Thus, this ELISA system showed that the three genera of molds, Aspergillus, Penicillium, or Fusarium, which are three major important molds producing mycotoxins in food or agricultural commodities, could be detected at the same time, using one detection system.

Development of a Functional Mortar for Restraining Surface Algal Growth

  • Park, Soon-young;Kim, Jinhyun;Kang, Hojeong
    • Ecology and Resilient Infrastructure
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    • 제5권2호
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    • pp.82-87
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    • 2018
  • Proliferation of algae on the surface of concrete or mortar in aquatic habitat has a negative impact on maintenance of concrete-based structures. Growth of algae may decrease stability of structure by bio-deterioration. In this study, we developed a functional mortar for restraining bio-deterioration by using $Cu^{2+}$ ion. The mortar contains soluble glass beads made of $Cu^{2+}$ ion, which can dissolve into water slowly. Mortars prepared with different ratio of glass beads (0, 2, 5, 10, and 15%) were placed in a culture medium with algae and incubated over a month period. Water chemistry, chlorophyll-a, and extracellular enzyme activities were measured. The incubation was conducted in both freshwater and seawater conditions, to assess applicability to both aquatic conditions. Overall, mortar with Cu glass exhibited lower chlorophyll-a content, suggesting that the functional mortar reduced algal growth. DOC concentration increased because debris of dead algae increased. Cu glass also decreased phosphatase activity, which is involved in the regeneration of inorganic P from organic moieties. Since, P is often a limiting nutrient for algal production, algal growth may be inhibited. Activities of ${\beta}$-glucosidase and N-acetylglucosaminidase were not significantly affected because carbon and nitrogen mineralization may not be influenced by the Cu glass beads. Our study suggests that functional mortar with Cu glass beads may reduce the growth of algae on the surface, while it has little environmental impact.

The role and characterization of .betha.-1, 3-glucanase in biocontrol of fusarium solani by pseudomonas stutzeri YPL-1

  • Lim, Ho-Seong;KiM, Sang-Dal
    • Journal of Microbiology
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    • 제33권4호
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    • pp.295-301
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    • 1995
  • An antifungal Pseudomonas stutzeri YPL-1 produced extracellular chitinase and .betha.-1, 3-glucanase that were key enzymes in the decomposition of fungal hyphal walls. These lytic extracellular enzymes markedly inhibited mycelial growth of the phytopathogenic fungus Fusarium solani. A chitinase from P. stutzeri YPL-1 inhibited fungal mycelial growth by 87%, whereas a .betha.-1, 3-glucanase from the bacterium inhibited growth by 53%. Furthermore, co-operative action of the enzymes synergistically inhibited 95% of the fungal growth. The lytic enzymes caused absnormal swelling and retreating on the fungal hyphal walls in a dual cultures. Scanning electron microscopy clearly showed hyphal degradation of F. solani in the regions interacting with P. stutzeri YPL-1. In an in vivo pot test, P. stutzeri YPL-1 proved to have biocontrol ability as a powerful agent in controlling plant disease. Planting of kidney bean (Phaseolus vulgaris L.) seedlings with the bacterial suspension in F. solani-infested soil significantly suppressed the development of fusarial root-rot. The characteristics of a crude preparation of .betha.-1, 3-glucanase produced from P. stutzeri YPL-1 were investigated. The bacterium detected after 2 hr of incubation. The enzyme had optimum temperature and pH of 40.deg.C and pH 5.5, respectively. The enzyme was stable in the pH range of 4.5 to 7.0 and at temperatures below 40.deg.C, with a half-life of 40 min at 60.deg.C.

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Differences in Biogeochemical Properties and Microbial Activities in Stream Segments with Changes in Land-use Type

  • Kim, Jinhyun;Jang, Inyoung;Lee, Hyunjin;Kang, Hojeong
    • Ecology and Resilient Infrastructure
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    • 제2권3호
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    • pp.247-254
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    • 2015
  • Changes in land-use type can affect soil and water properties in stream ecosystems. This study examined the effects of different land-use types on biogeochemical properties and microbial activities of a stream. We collected water and sediment samples in a stream at three different sites surrounded by varying land-use types; a forest, a radish field and a rice paddy. Nitrogen contents, such as nitrate, nitrite and total nitrogen in the stream water body, showed significant differences among the sampling sites. The highest nitrogen values were recorded at the site surrounded by cropland, as fertilizer runoff impacted the stream. Soil organic matter content in the sediment showed significant differences among sites, with the highest content exhibited at the forest mouth site. These differences might be due to the organic matter in surrounding terrestrial ecosystems. Microbial activities determined by extracellular enzyme activities showed similar values throughout all sites in the water body; however, the activities in the sediments exhibited the highest values near the forest site and mirrored the soil organic matter content values. From these results, we conclude that different land-use types are important factors affecting water and sediment properties in stream ecosystems.

Characterization of L-asparaginase-producing Trichoderma spp. Isolated from Marine Environments

  • Woon-Jong, Yu;Dawoon, Chung;Yong Min, Kwon;Seung Sub, Bae;Eun-Seo, Cho;Hye Suck, An;Grace, Choi
    • 한국해양생명과학회지
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    • 제7권2호
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    • pp.121-128
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    • 2022
  • L-asparaginase (ASNase) is a therapeutic enzyme used to treat acute lymphoblastic leukemia. Currently, the most widely used ASNases are originated from bacteria. However, owing to the adverse effects of bacterial ASNases, new resources for ASNase production should be explored. Fungal enzymes are considered efficient and compatible resources of natural products for diverse applications. In particular, fungal species belonging to the genus Trichoderma are well-known producers of several commercial enzymes including cellulase, chitinase, and xylanase. However, enzyme production by marine-derived Trichoderma spp. remains to be elucidated. While screening for extracellular ASNase-producing fungi from marine environments, we found four strains showing extracellular ASNase activity. Based on the morphological and phylogenetic analyses using sequences of translation elongation factor 1-alpha (tef1α), the Trichoderma isolates were identified as T. afroharzianum, T. asperellem, T. citrinoviride, and Trichoderma sp. 1. All four strains showed different ASNase activities depending on the carbon sources. T. asperellem MABIK FU00000795 showed the highest ASNase value with lactose as a carbon source. Based on our findings, we propose that marine-derived Trichoderma spp. are potential candidates for novel ASNase production.