• 제목/요약/키워드: Enzyme polymerization

검색결과 81건 처리시간 0.034초

Leuconostoc lactis CCK940의 Glucansucrase 활성에 의한 올리고당 생산 최적화 (Oligosaccharide Production by Leuconostoc lactis CCK940 Which Has Glucansucrase Activity)

  • 이설희;박영서
    • 산업식품공학
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    • 제21권4호
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    • pp.383-390
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    • 2017
  • 국내 재래시장에서 수집한 발효식품 등에서 우수한 glucansucrase 활성을 나타내는 유산균주를 분리한 후 이 균주를 이용한 올리고당 생성의 최적 조건을 조사하였다. 배추김치로부터 분리된 유산균주 CCK940은 glucansucrase 활성이 918.2 mU/mL로 가장 높아 본 올리고당 생산을 위한 균주로 선정하였고, Leu. lactis CCK940로 동정 및 명명되었다. 선정된 Leu. lactis CCK940는 배지의 pH를 6.0으로 조정하고 공여체인 sucrose와 수용체인 maltose의 초기 농도를 각각 5% (w/v)와 10% (w/v)로 첨가한 후 배양 4시간과 8시간째에 sucrose를 5% (w/v) 첨가하는 것이 최적인 것으로 확인되었다. 최적 조건에 12시간 배양 시 Leu. lactis CCK940는 중합도가 2-4인 올리고당을 최소 4종류 생성하였다. 본 균주는 수용체 분자로서 fructose와 melibiose를 사용할 수 없었다.

Echinococcus granulosus Protoscolex DM9 Protein Shows High Potential for Serodiagnosis of Alveolar Echinococcosis

  • Kim, Jeong-Geun;Han, Xiumin;Kong, Yoon
    • Parasites, Hosts and Diseases
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    • 제60권1호
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    • pp.25-34
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    • 2022
  • Alveolar echinococcosis (AE) caused by infection with E. multilocularis metacestode, represents one of the most fatal helminthic diseases. AE is principally manifested with infiltrative, proliferating hepatic mass, resembling primary hepatocellular carcinoma. Sometimes metastatic lesions are found in nearby or remote tissue. AE diagnosis largely depends on imaging studies, but atypical findings of imaging features frequently require differential diagnosis from other hepatic lesions. Serological tests may provide further evidence, while obtaining reliable AE materials is not easy. In this study, alternative antigens, specific to AE were identified by analyzing E. granulosus protoscolex proteins. An immunoblot analysis of E. granulosus protoscolex showed that a group of low-molecular-weight proteins in the range from 14 kDa to 16 kDa exhibited a sensitive and specific immune response to AE patient sera. Partial purification and proteomic analysis indicated that this protein group contained myosin, tubulin polymerization promoting protein, fatty-acid binding protein, uncharacterized DM9, heat shock protein 90 cochaperone tebp P-23, and antigen S. When the serological applicability of recombinant forms of these proteins was assessed using enzyme-linked immunosorbent assay, DM9 protein (rEgDM9) showed 90.1% sensitivity (73/81 sera tested) and 94.5% specificity (172/181 sera tested), respectively. rEgDM9 showed weak cross-reactions with patient sera from the transitional and chronic stages of cystic echinococcosis (3 to 5 stages). rEgDM9 would serve as a useful alternative antigen for serodiagnosis of both early- and advanced-stage AE cases.

Quercetin Reduces Chemotactic Activity of Porcine Peripheral Blood Polymorphonuclear Cells

  • Hwa, Gyeong-Rok;Ahn, Changhwan;Kim, Hakhyun;Kang, Byeong-Teck;Jeung, Eui-Bae;Yang, Mhan-Pyo
    • 한국임상수의학회지
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    • 제39권2호
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    • pp.51-58
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    • 2022
  • Quercetin, a flavonoid found in fruits and vegetables, exhibits a strong anti-inflammatory activity. The objective of this study was to examine the effect of quercetin on chemotactic activity of peripheral blood polymorphonuclear cells (PMNs) to culture supernatant from peripheral blood mononuclear cells (PBMCs) stimulated with lipopolysaccharide (LPS). In addition, we determined whether this effect is related to interleukin (IL)-8 and changes in cytoskeleton. The chemotactic activity of PMNs was evaluated by a modified Boyden chamber assay. Total cellular filamentous (F)-actin levels were measured by method of fluorescence microscopy. The levels of IL-8 mRNA and protein were measured by real time polymerase reaction method and enzyme-linked immunosorbent assay, respectively. Quercetin (0-50 µM) itself has no chemoattractant effect for PMNs. The culture supernatant from PBMCs (2 × 106 cells/mL) treated with LPS (1 ㎍/mL) showed remarkable increase in chemotaxis of PMNs. However, this effect was reduced dose-dependently by treatment with quercetin. In addition, PBMCs treated with LPS revealed enhanced levels in IL-8 protein and mRNA. Co-treatment of LPS with quercetin (50 µM) in PBMCs decreased IL-8 production and expression. Treatment of quercetin (0-50 µM) on PMNs to rpIL-8 (10 nM) decreased dose-dependently the chemotactic activity of PMNs. Treatment of quercetin on PMNs to IL-8 also reduced their total cellular F-actin level. These results suggested that quercetin attenuates chemotactic activity of PMNs, which is mediated by down-regulation of IL-8 production from LPS-stimulated PBMCs and inhibition of F-actin polymerization in PMNs.

Xylogone sphaerospora 유래 ${\beta}$-mannanase 정제 및 Konjac Glucomannan 가수분해 올리고당의 중합도별 Bifidobacterium spp.에 대한 증식활성 (Purification of Xylogone sphaerospora ${\beta}$-mannanase and Growth Activity of Bifidobacterium spp. by Konjac Glucomannan Hydrolysates)

  • 이희정;박귀근
    • Applied Biological Chemistry
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    • 제51권3호
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    • pp.159-163
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    • 2008
  • Sephadex G-100 column chromatography에 의해 Xylogone sphaerospora 유래 ${\beta}$-mannanase의 정제를 수행하여 비활성 8.44 units/ml, 정제배율 56.27을 나타내었다. SDS-PAGE에 의한 단일밴드를 확인하였고, 분자량은 42kDa으로 결정되었다. 정제효소에 의해 konjac glucomannan을 가수분해하여 activated carbon column chromatograph)에 의해 당가수분해물을 분리 회수하여 TLC에 의해 주요 당가수분해물은 합도 3과 4의 hetero type으로 확인되었으며 D.P. 3과 4의 예상되는 구조는 본 연구실에서 확보하고 있는 standard glucomannooligosaccharides에 의한 TLC에서 나타나는 Rf value 상으로 1차적으로 확인하고, Aspergillus niger 5-16 유래 정제 ${\beta}$-mannanase를 단계적으로 처리하여 가수분해 pattern을 TLC로 해석한 결과 D.P. 3의 구조식은 비환원말단 mannose로 부터 2번째에 1분자의 glucose가 결합하고 있는 hetero type의 구조(M-G-M)로, D.P. 4의 구조식은 비환원말단 mannose로부터 3번째에 1분자의 glucose가 결합하고 있는 hetero type의 구조(M-M-G-M)로 예상하고 있다. B. longumm, B. bifidum, B. infantis, B. adolescentis, B. animalis, B. auglutum, B. breve의 생육활성에 대한 중합도 3과4의 영향을 검토하기 위하여 modified-MRS 배지 상에 탄소원으로 중합도 3과 4를 대체하여 생육활성을 비교한 결과 B. longum에서는 D.P 4 glucomannooligosaccharide를 탄소원으로 대체한 경우 표준 MRS배지와 비교하여 3.9배의, D.P. 3을 처리한 경우에도 2.7배의 상대 활성을 나타내어 가장 우수한 생육활성을 나타냈었으며, B. breve의 경우에서도 D.P 4에서 2.47배, D.P 3에서 2.08배의 활성을 나타내었으며 B. bifidum에 있어서는 D.P. 4의 경우 2.8배의 상대활성을 나타내었다. Bifidobacterium 7균주 모두에 대해서 중합도 4의 올리고당이 중합도 3의 올리고당보다 생육활성에 크게 기여하는 것으로 나타났다.

무수치 표백술 후 잔존 과산화수소수 제거를 위한 수종의 치수강 세척제의 효과에 관한 정량적 연구 (A QUANTITATIVE STUDY ON THE DEGRADING EFFECT OF THE VARIOUS IRRIGATING AGENTS IN THE ELIMINATION OF RESIDUAL HYDROGEN PEROXIDE FOLLOWING WALKING BLEACHING)

  • 금기연;한원섭;정일영;이승종;이찬영;오병훈
    • Restorative Dentistry and Endodontics
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    • 제23권2호
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    • pp.656-669
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    • 1998
  • Hydrogen peroxide at high concentration during walking bleaching may cause damage to the tooth structure and to the surrounding periodontal tissues and may develop external root resorption. Clinically, It is so important to find a method of prevention or minimization of these complications. The efficacy of various chamber-irrigating agents to eliminate residual hydrogen peroxide after walking bleaching was examined and compared with water rinse in this study. Extracted human 46 premolars without any cementoenamel junction defects were treated endodontically and based with IRM to 1 mm below CEJ and totally bleached 3 times for each tooth with 30% hydrogen peroxide and sodium perborate. Upon completion of the 3rd walking bleaching procedure, the cervical portion and pulp chamber of each group of teeth were irrigated with catalase, 70% ethylalcohol, acetone, and distilled water. And then, a radicular hydrogen peroxide penetration was measured with spectrophotometer immediately after each bleaching and following treatment with each chamber-irrigating agents, and the significance of their eliminating efficacy of residual hydrogen peroxide was analyzed by Kruskal-Wallis test. The results were obtained as follows. 1. Cervical root penetration of hydrogen peroxide was increased as the bleaching procedure was repeated(P<.01). 2. The most effective irrigant that removed residual hydrogen peroxide was the catalase, and the least effective one was water rinsing (P<.01).; there was no significant difference between the acetone and ethanol group. 3. The Irrigation with antioxidant enzyme or water-displacement solutions can eliminate residual oxygen radicals from the pulp chamber effectively after walking bleaching. So, these agents can reduce adverse effects such as cervical external resorption and periapical inflammation and prevent residual $O_2$ from impeding composite resin polymerization, thus increase the bonding strength of composite resin. This, in turn reduces microleakage and discoloration of the esthetic restoration, extending its service-life.

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Purification and Characterization of a Thermostable Xylanase from Paenibacillus sp. NF1 and its Application in Xylooligosaccharides Production

  • Zheng, Hong-Chen;Sun, Ming-Zhe;Meng, Ling-Cai;Pei, Hai-Sheng;Zhang, Xiu-Qing;Yan, Zheng;Zeng, Wen-Hui;Zhang, Jing-Sheng;Hu, Jin-Rong;Lu, Fu-Ping;Sun, Jun-She
    • Journal of Microbiology and Biotechnology
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    • 제24권4호
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    • pp.489-496
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    • 2014
  • High levels of extracellular xylanase activity (211.79 IU/mg) produced by Paenibacillus sp. NF1 were detected when it was submerged-cultured. After three consecutive purification steps using Octyl-Sepharose, Sephadex G75, and Q-Sepharose columns, a thermostable xylanase (XynNF) was purified to homogeneity and showed a molecular mass of 37 kDa according to SDS-PAGE. The specific activity of the purified XynNF was up to 3,081.05 IU/mg with a 14.55-fold purification. The activity of XynNF was stimulated by $Ca^{2+}$, $Ba^{2+}$, DTT, and ${\beta}$-mercaptoethanol, but was inhibited by $Fe^{2+}$, $Zn^{2+}$, $Fe^{2+}$, $Cu^{2+}$, SDS, and EDTA. The purified XynNF displayed a greater affinity for oat spelt xylan with the maximal enzymatic activity at $60^{\circ}C$ and pH 6.0. XynNF, which was shown to be cellulose-free, with high stability at high temperature ($70^{\circ}C-80^{\circ}C$) and low pH range (pH 4.0-7.0), is potentially valuable for various industrial applications. The enzyme hydrolyzed oat spelt xylan to yield mainly xylooligosaccharides (95.8%) of 2-4 degree of polymerization (DP2-4). Moreover, the majority of the xylooligosacharides (DP2-4) products was xylobiose (61.5%). The thermostable xylanase (XynNF) thus seems potentially usefull in the production of xylooligosaccharides.

미생물 Transglutaminase를 이용하여 제조된 쌀 혼합 전두부의 이화학적 및 물성 평가 (Physicochemical and Rheological Evaluation of Rice-Whole Soybean Curds Prepared by Microbial Transglutaminase)

  • 진익훈;이삼빈
    • 한국식품영양과학회지
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    • 제40권5호
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    • pp.738-746
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    • 2011
  • Microbial transglutaminase(MTGase)를 이용하여 쌀 가수분해물과 초미세 생대두 분말을 혼합하여 전두부를 제조한 후 조직감, 동적점탄성, 단백질 결합패턴 및 미세구조를 평가하였다. 40%(w/v) 쌀 용액을 Termamyl 효소처리($85^{\circ}C$, 20분) 하였을 때 점조도 값 및 환원당 함량이 각각 $1.27\;Pa{\cdot}s^n$, 9.0%로 나타났다. MWSP 18~22% 농도에서 MTGase 효소로 응고시킨 전두부의 물성을 측정한 결과 MWSP 22% 첨가구에서 전형적인 두부의 조직감을 얻었으며, 쌀 가수분해물 7.5% 첨가구의 경우 경도 639.6 dyne/$cm^2$, 탄성 0.96으로 나타났다. MTGase 5%의 첨가 조건에서 MWSP 18~22% 농도에 따라 동적점탄성을 측정한 결과 MWSP 22% 첨가 경우 반응 6분 이내 G'(5.1 Pa) 및 G''(9.0 Pa)값으로 높게 나타났다. MWSP 22% 첨가구에서 시간에 따른 SDS-PAGE 상에서 대부분의 콩 단백질은 30분 이내에 중합되어 고분자중합체를 형성하였으며, 쌀 가수분해물이 첨가된 경우에도 대부분의 콩 단백질들이 중합되는 유사한 경향을 보였다. 쌀이 첨가된 전두부의 미세구조는 균일한 입자로 채워진 네트워크 구조를 보였으나 냉장저장 2일 후에는 쌀을 첨가하지 않은 대조구에 비해 더욱 거칠어진 표면을 확인할 수 있었다.

E. coli lipopolysaccharides로 유도된 사람 호중구에서 CD14, Toll-like receptors, cytoskeletal inhibitors 그리고 $NF-{\kappa}B$ inhibitor가 MMP-8 분비에 미치는 영향 (Effect of CD14, Toll-like receptors, cytoskeletal inhibitors and $NF-{\kappa}B$ inhibitor on MMP-8 release from human neutrophils induced by E. coli lipopolysaccharides.)

  • 양승민;김태일;설양조;이용무;구영;정종평;한수부;류인철
    • Journal of Periodontal and Implant Science
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    • 제35권2호
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    • pp.427-436
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    • 2005
  • Objective: MMP-8 is a neutrophil enzyme and its level increases in some inflammatory diseases, including periodontal disease. We knew that the lipopolysaccharide of E.coli(E-LPS) induced MMP-8 release from human neutrophils. E-LPS is known to induce the production and release of inflammatory cytokines through CD14, Toll-like receptor(TLR). In the present study, we investigated whether MMP-8 release by E-LPS is induced via CD14-TLR pathway and the cellular mechanism of MMP-8 release in human neutrophils. Material and methods: Human neutrophils were isolated from the peripheral blood of healthy donors and pre-incubated in medium containing antibodies against CD14, anti-TLR2 and anti-TLR4 or several inhibitors of microtubules and microfilaments and then incubated with E-LPS. The cells were treated TPCK and E-LPS simultaneously. The MMP-8amount in the culture medium was determined using ELISA. Results: E-LPS increased MMP-8release from neutrophils and its induction was inhibited by anti-CD14 and anti-TLR4 but not by anti-TLR2 antibodies. The inhibitors of microtubule and microfilament polymerization significantly decreased E-LPS-induced MMP-8release. TPCK inhibited E-LPS-induced MMP-8 release. Conclusion: These results suggest that MMP-8 release is induced by E-LPS via the CD14-TLR4 signal pathway in human neutrophils and may be depedent on microtubule and microfilament systems and $NF-{\kappa}B$ pathway.

Bacillus sp.유래 $\beta$-Mannanase 정제 및 Guar Gum가수분해 올리고당의 Bifidobacterium spp.에 대한 증식활성 (Purification of Bacillus sp. $\beta$-Mannanase and the Growth Activity of Bifidobacterium spp. by Guar Gum Hydrolysates.)

  • 최준영;박귀근
    • 한국미생물·생명공학회지
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    • 제32권2호
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    • pp.117-122
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    • 2004
  • DEAE-sephadex ion exchange column chromatography에 의해 Bacillus sp. 유래 $\beta$-mannanase 의정제를 수행하여 비활성 21.57 units/$m\ell$, 정제배율 95.33을 나타내었다. SDS-PAGE에 의한 단일밴드를 확인하였고, 분자량은 38.9 kDa으로 결정되었다. 정제효소에 의해 guar gum galactomannan을 가수분해하여 1차 activated carbon column chromatography 와 2차 Sephadex G-25 gel filtration에 의해 당가수분해물을 분리 회수하여 TLC 및 FACE 에 의해 주요 당가수분해물은 중합도 5와 7로 확인되었다. B. longum B. bifidum, B. infantis, B. adolescentis, B. animalis, and B. breve의 생육활성에 대한 중합도 5와 7의 영향을 검토하기 위하여 modified-MRS 배지상에 탄소원으로 중합도 5와 7을 대체하여 생육활성을 비교한 결과 B. longum 에서는 중합도 5 galactomannooligosaccharide를 탄소원으로 대체한 경우 Standard MRS와 배교하여 0.62배로 감소하였다. 또한 중합도 5의 올리고당이 중합도 7의 올리고당보다 생육활성에 크게 기여하는 것으로 나타났다.

Inflammatory Effect of Light-Emitting Diodes Curing Light Irradiation on Raw264.7 Macrophage

  • Jeong, Moon-Jin;Kil, Ki-Sung;Lee, Myoung-Hwa;Lee, Seung-Yeon;Lee, Hye-Jin;Lim, Do-Seon;Jeong, Soon-Jeong
    • 치위생과학회지
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    • 제19권2호
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    • pp.133-140
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    • 2019
  • Background: The light-emitting diode (LED) curing light used is presumed to be safe. However, the scientific basis for this is unclear, and the safety of LED curing light is still controversial. The purpose of this study was to investigate the effect of LED curing light irradiation according to the conditions applied for the polymerization of composite resins in dental clinic on the cell viability and inflammatory response in Raw264.7 macrophages and to confirm the stability of LED curing light. Methods: Cell viability and cell morphology of Raw264.7 macrophages treated with 100 ng/ml of lipopolysaccharide (LPS) or/and LED curing light with a wavelength of 440~490 nm for 20 seconds were confirmed by methylthiazolydiphenyl-tetrazolium bromide assay and microscopic observation. The production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) was confirmed by NO assay and $PGE_2$ enzyme-linked immunosorbent assay kit. Expression of interleukin $(IL)-1{\beta}$ and tumor necrosis factor $(TNF)-{\alpha}$ in total RNA and protein was confirmed by reverse transcription polymerase chain reaction and Western blot analysis. Results: The LED curing light did not affect the viability and morphology of normal Raw264.7 cells but affected the cell viability and induced cytotoxicity in the inflammation-induced Raw264.7 cells by LPS. The irradiation of the LED curing light did not progress to the inflammatory state in the inflammation-induced Raw264.7 macrophage. However, LED curing light irradiation in normal Raw264.7 cells induced an increase in NO and $PGE_2$ production and mRNA and protein expression of $(IL)-1{\beta}$ and $(TNF)-{\alpha}$, indicating that it is possible to induce the inflammatory state. Conclusion: The irradiation of LED curing light in RAW264.7 macrophage may induce an excessive inflammatory reaction and damage oral tissues. Therefore, it is necessary to limit the long-term irradiation which is inappropriate when applying LED curing light in a dental clinic.