• 제목/요약/키워드: 1,3-glucanase

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색소에 접합된 $\beta$-glucan을 이용한 $\beta$-glucan 분해효소 생산 균주의 분리 및 동정 (Isolation and identification of $\beta$-glucan degrading enzyme producing bacterium using coloured $\beta$-glucan)

  • 양진오;정안식;이성택
    • 미생물학회지
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    • 제25권4호
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    • pp.339-345
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    • 1987
  • A bacterium K-4-3, producing $\beta$-glucan hydrolyzing enzyme, was isolated from soil and identified to be Bacillus subtilis by its morpholohical and physiological characteristics. $\beta$-glucan was coloured using cibacron blue 3G-A and cross linded by the addition of 1, 4-butanedioldiglycidyl ether. This substrate was used for the isolation of $\beta$-glucanase producing microorganism. The $\beta$-glucan hydrolyzing enzyme actibity from isolated K-4-3 strain was also measured using the modified substrate. Bacillus subtilis K-4-3 produced the highest extracellular $\beta$-glucan hydrolyzing activity in the basal medium containing $\beta$-glucan as a carbon source, peptone and tryptone as a nitrogen source, and magnesium sulfate as an inorganic salt. The optimum temperature and initial pH for $\beta$-glucanase production by Bacillus subtilis K-4-3 were $37^{\circ}C$ and pH6. The highest enzyme activity was obtained at the culture age of 54 hrs with rotary shaking at $37^{\circ}C$. The crude enzyme showed the highest activity at pH 7.5-8.0 and $65^{\circ}C$.

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지렁이 중장에서 발현되는 Endo-$\beta$-1,4-glucanase의 동정 및 특성에 관한 연구 (Isolation and Characterization of Endo-$\beta$-1,4-glucanase from the Midgut of the Earthworm, Eisenia andrei)

  • 이명식;조성진;탁은식;허소영;이종애;박범준;조현주;신주옥;박순철
    • 한국토양동물학회지
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    • 제8권1_2호
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    • pp.7-12
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    • 2003
  • 지렁이 (Eisenia andrei)의 중장에서 내생의 endoglucanase 유전자의 전체 염기 서열을 동정하였다. ORF의 길이는 1,371 bp이며, 456개의 아미노산으로 번역된다. NCBI에 등록된 가재와 흰개미의 cellulase 및 endo-$\beta$-1, 4-glucanase와 50-51%의 유사성을 보이며, 활성 부위가 잘 보존되어 있었다. 계통수 분석에서는 다른 동물 분류군에서 밝혀진 GHF9 그룹의 cellulase와 근연관계가 없음이 확인되었다.

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Roles of Carbohydrate-Binding Module (CBM) of an Endo-β-1,4-Glucanase (Cel5L) from Bacillus sp. KD1014 in Thermostability and Small-Substrate Hydrolyzing Activity

  • Lee, Jae Pil;Shin, Eun-Sun;Cho, Min Yeol;Lee, Kyung-Dong;Kim, Hoon
    • Journal of Microbiology and Biotechnology
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    • 제28권12호
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    • pp.2036-2045
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    • 2018
  • An endo-${\beta}$-1,4-glucanase gene, cel5L, was cloned using the shot-gun method from Bacillus sp.. The gene, which contained a predicted signal peptide, encoded a protein of 496 amino acid residues, and the molecular mass of the mature Cel5L was estimated to be 51.8 kDa. Cel5L contained a catalytic domain of glycoside hydrolase (GH) family 5 and a carbohydrate-binding module family 3 (CBM_3). Chromatography using HiTrap Q and CHT-II resulted in the isolation of two truncated forms corresponding to 50 (Cel5L-p50) and 35 kDa (Cel5L-p35, CBM_3-deleted form). Both enzymes were optimally active at pH 4.5 and $55^{\circ}C$, but had different half-lives of 4.0 and 22.8 min, respectively, at $70^{\circ}C$. The relative activities of Cel5L-p50 and Cel5L-p35 for barley ${\beta}$-glucan were 377.0 and 246.7%, respectively, compared to those for carboxymethyl-cellulose. The affinity and hydrolysis rate of pNPC by Cel5L-p35 were 1.7 and 3.3 times higher, respectively, than those by Cel5L-p50. Additions of each to a commercial enzyme set increased saccharification of pretreated rice straw powder by 17.5 and 21.0%, respectively. These results suggest CBM_3 is significantly contributing to thermostability, and to affinity and substrate specificity for small substrates, and that these two enzymes could be used as additives to enhance enzymatic saccharification.

토양(土壤)에서 분리(分離)한 Streptomyces sp. s-45의 효소학적(酵素學的) 성질(性質)에 관한 연구(硏究) (Studies on the Enzymatical Properties of Streptomyces sp. S-45 Isolated from Soil)

  • 김영일;김용웅;김광식
    • 한국토양비료학회지
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    • 제21권2호
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    • pp.129-134
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    • 1988
  • 토양(土壤)으로부터 분리(分離)한 Streptomyces sp. S-45 균주(菌株)의 효소학적(酵素學的) 성질(性質)을 검토(檢討)한 결과(結果)는 다음과 같다. 1. 분리균(分離菌) Streptomyces sp. S-45의 Chitinase activity는 $3.01({\mu}/m{\ell})$이고 ${\beta}$-1.3-Glucanase activity는 $2.49({\mu}/m{\ell})$이었다. 2. 탄소원(炭素原)으로서 Colloidal chitin 0.7%, Glucose 0.3%, 질소원(窒素原)으로서 Asparagine 0.5%, Peptone 0.2% 존재(存在)가 효소생산(酵素生産)에 효과적(效果的)이었다. 3. 최적(最適) 효소생성(酵素生成) 조건(條件)으로 pH 7.0, $30^{\circ}C$에서 6일간(日間) 진탕배양시(培養時) 최고(最高)의 효소생성(酵素生成)을 나타내었다. 4. 효소(酵素)의 활성(活性)에 미치는 최적(最適) 작용조건(作用條件)은 pH 6.5~7.0, 온도(溫度)는 $45{\sim}50^{\circ}C$였다. 5. 본(本) 효소(酵素)는 pH6.0~7.0에서 안정성(安定性)이 가장 높았고, $80^{\circ}C$로 10분(分) 처리시(處理時) Chitinase는 10%, ${\beta}$-1.3-Glucanase는 12%로 잔존활성(殘存活性)이 감소(減少)하였다. 6. 금속(金屬)이온에 대한 효소(酵素)의 영향(影響)은 $Co^{{+}{+}}$, $Cu^{{+}{+}}$, $Mn^{{+}{+}}$, $Al^{{+}{+}{+}}$$10^{-2}M$$Sn^{{+}{+}}\;10^{-3}M$에서 활성(活性)이 증대(增大)하였고, $Ag^{{+}{+}}$, $Hg^{{+}{+}}$는 현저(顯著)한 저해작용을 하였다.

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Pilot-Scale Production of Cellulase Using Trichoderma reesei Rut C-30 Fed-Batch Mode

  • Lee, Sang-Mok;Koo, Yoon-Mo
    • Journal of Microbiology and Biotechnology
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    • 제11권2호
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    • pp.229-233
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    • 2001
  • Trichoderma reesei Rut C-30 produced high levels of ${\beta}$-glucosidase, endo-${\beta}$-glucosidase, endo-${\beta}$-1,4-glucanase, and exo-${\beta}$-1,4-glucanase. In pilot-scale production (50-1 fermentor), productivity and yield of CMCase (carborymethyl cellulose) and FPase (filter paper activity) were 273 U/ml and 35 U/ml, and 162 FPU/l.h and 437 FPU/g, respectively. The fed-batch techniques were used to improve enzyme activities with constant cell concentration. The acidity was an important parameter and controlled at pH 3.9 and 5.0 by automatic addition of ammonium hydroxide. Cellulase powder was prepared by ammonium sulfate precipitation and its CMCase and FPase activities were 3,631 U/g and 407 U/g, respectively.

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Effect of Acibenzolar-S-methyl and Rahnella aquatilis (Ra39) on Chitinase and β-1, 3-glucanase Activities and Disease Resistance of Apple Plants

  • Abo-Elyousr, A.M. Kamal;Sallam, M.A.A.;Hassan, M.H.A.;Zeller, W.
    • The Plant Pathology Journal
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    • 제26권1호
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    • pp.63-69
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    • 2010
  • The effect of Acibenzolar-S-methyl (ASM) and Rahnella aquatilis Ra39 against apple fire blight disease caused by Erwinia amylovora were tested as a possible alternative to streptomycin. In vitro studies, no inhibition effect against the pathogen was found when ASM was tested. Under greenhouse conditions, application of R. aquatilis Ra39 with the highly susceptible M26 rootstock resulted in a marked disease suppression. Application of ASM and strain Ra39 caused a high decrease of the disease, 82% and 58% respectively; this was correlated with a reduction of the growth of the pathogen within host plants up to 64% and 49.5% respectively. Further studies in the field under artificial infection condition during full bloom revealed that application of ASM and R. aquatilis Ra39 with Gala variety resulted in a control effect up to 21 and 29% respectively. In physiological studies, enhanced activities of PR-proteins (chitinase and $\beta$-1, 3-glucanase) were detected, which are well known as biochemical markers for systemic acquired resistance. Application of ASM to apple shoots caused the highest chitinase activity followed by strain Ra39. The enzyme activity was increased after 2, 4 and 6 days from application. In addition, ASM-treatment caused the higher $\beta$-1, 3-glucanase activity than strain Ra39. Maximum enzyme activity was recorded after 6 days from application and then decreased after 8 and 10 days from application.

지렁이 중장에서 발현되는 endo-β-1,4-glucanase 유전자들의 클로닝과 특성에 관한 연구 (Cloning and Characterization of endo-β-1,4-glucanase genes from the Midgut of the Earthworm, Eisenia andrei)

  • 이명식;박상길;탁은식;안치현;김혜령;박순철
    • 유기물자원화
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    • 제15권3호
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    • pp.80-89
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    • 2007
  • 셀룰로오스 가수분해효소의 하나인 endo-${\beta}$-1,4-D-glucanase의 유전자를 지렁이 Eisenia anderi의 중장으로부터 클로닝하여 EaEG2와 EaEG3로 명명하였다. 두 유전자의 염기는 1368bp이며, 개시코돈을 포함하여 456개의 아미노산을 코딩한다. N-말단 지역의 20개 잔기들은 signal peptide이다. 두 유전자의 전체 아미노산 염기서열은 glycosyl hydrolase family 9에 속하며, 같은 종류의 셀룰로오스 가수분해효소를 분비하는 흰개미, 바퀴벌레, 가재 그리고 연체동물과 51-55%의 높은 상동성을 보였다. 지렁이의 EaEG2와 EaEG3는 가수분해활성에 중요한 세 부분이 잘 보존되어 있다. 아미노산 염기서열을 이용한 계통수 분석에서 GHF9 그룹은 절지동물, 박테리아, 식물, 환형동물 및 연체동물의 5개 그룹으로 분석되었다.

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Biocontrol of Anthracnose in Pepper Using Chitinase, ${\beta}$-1,3 Glucanase, and 2-Furancarboxaldehyde Produced by Streptomyces cavourensis SY224

  • Lee, So Youn;Tindwa, Hamisi;Lee, Yong Seong;Naing, Kyaw Wai;Hong, Seong Hyun;Nam, Yi;Kim, Kil Yong
    • Journal of Microbiology and Biotechnology
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    • 제22권10호
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    • pp.1359-1366
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    • 2012
  • A strain of Streptomyces cavourensis subsp. cavourensis (coded as SY224) antagonistic to Colletotrichum gloeosporioides infecting pepper plants was isolated. SY224 produced lytic enzymes such as chitinase, ${\beta}$-1,3-glucanase, lipase, and protease in respective assays. To examine for antifungal activity, the treatments amended with the nonsterilized supernatant resulted in the highest growth inhibition rate of about 92.9% and 87.4% at concentrations of 30% and 10%, respectively. However, the sterilized treatments (autoclaved or chloroform treated) gave a lowered but significant inhibitory effect of about 63.4% and 62.6% for the 10% supernatant concentration, and 75.2% and 74.8% for the of 30% supernatant concentration in the PDA agar medium, respectively, indicative of the role of a non-protein, heat stable compound on the overall effect. This antifungal compound, which inhibited spore germination and altered hyphal morphology, was extracted by EtOAc and purified by ODS, silica gel, Sephadex LH-20 column, and HPLC, where an active fraction was confirmed to be 2-furancarboxaldehyde by GS-CI MS techniques. These results suggested that SY224 had a high potential in the biocontrol of anthracnose in pepper, mainly due to a combined effect of lytic enzymes and a non-protein, heat-stable antifungal compound, 2-furancarboxaldehyde.