• 한국미생물·생명공학회지
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• 제24권1호
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• pp.9-18
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• 1996

Streptococcus mutans has been implicated as primary causative agents of dental caries by insoluble glucan (IG) in human and experimental animals. An attempt was made to search for the ${\alpha}$-1,3 glucanase that degrades IG produced by S. mutans. ${\alpha}$-1,3 glucanase was detected in the culture supernatant of microorganisms, which are isolated from soils on agar medium containing IG as a sole carbon source. This Streptomyces sp. hydrolysed IG produced by immobilized S. mutans and was named as Y9373. This enzyme required ${\alpha}$-1,3 glucan (IG) as an inducer. The optimum conditions for enzyme production were studied. The enzyme was purified by 30~70% $(NH_4)_2SO_4$ precipitation, anion exchange chroma tography on DEAE-cellulose and gel filtration on Sepadex G-75. The purified enzyme has a specific activity of 7840.0 U/mg protein giving 32.1-fold purification and final yield of 0.53%. The molecular weight was estimated to be about 22.5 kDa by SDS-PAGE. The optimum pH and temperature for enzyme reaction were 6.5 and 37$^{\circ}C$, respectively and the enzyme was relatively stable at the temperature below 60$^{\circ}C$. The activity of purified enzyme was enhanced by adding $Co^{2+},\;Mn^{2+}\;and\;Mg^{2+}$ into the medium, whereas inhibited by adding $Hg^{2+},\;Zn^{2+}$ and SDS. The $K_m\;and\;V_{max}$ value of ${\alpha}$-1,3 glucanase for IG were estimated to be 2.50 mM and 0.0431 mM/min, respectively. The thin layer chromatographic analysis of hydrolysates from IG with ${\alpha}$-1,3 glucanase showed that glucose was the main product of reaction. This enzyme activity was about 14 times higher than marketing dextranase as preventive agent against artificial dental caries by S. mutans in TH medium including 5% sucrose after 30 minutes.

• 미생물학회지
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• 제40권3호
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• pp.230-231
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• 2004

꽃송이버섯(Sparassis crispa DSMZ 5201)의 균사를 사용하여 균사외 효소활성을 측정하였다. Yeast-malt extract-glucose 배지를 사용하여 $24^{\circ}C$에서 15일간 배양 후 배양여액을 조효소원으로 사용하였을 때, $\alpha$-amylase효소의 활성은 44.27 unit/$mg{\cdot}protein$이었다. 배양여액 중의 Protease, CMCase, $\beta$-glucosidase, chitinase 및 exo-$\beta$-1,4-glucanase의 세포외 효소활성은 상대적으로 높았으나, xylanase 효소활성은 낮게 나타났다.

• 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.

• 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 $Co^{{+}{+}}$, $Cu^{{+}{+}}$, $Mn^{{+}{+}}$, $Al^{{+}{+}{+}}$의 $10^{-2}M$과 $Sn^{{+}{+}}\;10^{-3}M$에서 활성(活性)이 증대(增大)하였고, $Ag^{{+}{+}}$, $Hg^{{+}{+}}$는 현저(顯著)한 저해작용을 하였다.

• Journal of Microbiology and Biotechnology
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• 제16권7호
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• pp.1132-1138
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• 2006

Three types of xylanases (EC 3.2.1.8) were detected in the strain Aspergillus niger A-25, one of which, designated as XynIII, also displayed ${\beta}-(l,3-1,4)-glucanase$ (EC 3.2.1.73) activity, as determined by a zymogram analysis. XynIII was purified by ultrafiltration and ion-exchange chromatography methods. Its apparent molecular weight was about 27.9 kDa, as estimated by SDS-PAGE. The purified XynIII could hydrolyze birchwood xylan, oat spelt xylan, lichenin, and barley ${\beta}-glucan$, but not CMC, avicel cellulose, or soluble starch under the assay conditions in this study. The xylanase and ${\beta}-(l,3-1,4)-glucanase$ activities of XynIII both had a similar optimal pH and pH stability, as well as a similar optimal temperature and temperature stability. Moreover, the effects of metal ions on the two enzymatic activities were also similar. The overall hydrolytic rates of XynIII in different mixtures of xylan and lichenin coincided with those calculated using the Michaelis-Menten model when assuming the two substrates were competing for the same active site in the enzyme. Accordingly, the results indicated that XynIII is a novel bifunctional enzyme and its xylanase and ${\beta}-(l,3-1,4)-glucanase$ activities are catalyzed by the same active center.

• 한국축산식품학회지
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• 제27권4호
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• pp.409-415
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• 2007

본 연구에서는 Ross 208 종의 broiler 31,800수를 대조구와 복합 효소제 첨가구로 나누어 5주간에 걸쳐 대조구는 옥수수-대두박을 기초로 한 시판 육계사료를 급여하고, 시험구는 ${\beta}-glucanase$를 강화한 복합 효소제를 0.3% 첨가 급여하여 생산성과 혈청성분과 육질에 미치는 영향을 조사하였다. 시험기간 동안의 각 시험구의 증체량 및 사료 효율, 생산지수, 육성율을 측정하였고 무작위로 각 시험구당 20수를 선발하여 혈청 콜레스테롤 및 glucose 농도를 측정하였다. 또 ND와 IBD 항원을 음용 투여한 브로일러의 ND와 IBD에 대한 항체역가를 조사하였으며, 육질검사로 도체의 pH 및 육색, 가열감량, 보수력, 전단력을 측정하였다. 사육시험 결과, 0.3% 복합 효소제 시험구가 대조구에 비해 평균체중 19%, 생산지수 6.8%, 사료효율 5.5%, 육성율 5%의 증가를 나타내었다. 복합 효소제 시험구는 대조구보다 혈청내 HDL-콜레스테롤 양을 증가시키고 glucose의 양을 감소시켰다. 육계 혈청 내 ND 및 IBD 항원에 대한 항체역가도 복합 효소제 시험구가 대조구보다 높았다. 육질검사에 있어서는 복합 효소제 시험구가 대조구보다 육색과 전단력은 차이가 없었으나 보수력(p<0.05)과 가열감량이 개선되는 결과를 나타내었다. 이상의 결과로부터 ${\beta}-glucanase$ 활성을 강화한 복합 효소제의 첨가 급여는, 육계의 생산성과 혈청성분과 면역기능과 육질을 개선시키는 효과가 있는 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제22권3호
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• pp.407-415
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• 2012

An endophytic bacterium, Bacillus thuringiensis GS1, was isolated from bracken (Pteridium aquilinum) and found to have maximal production of chitinase (4.3 units/ml) at 5 days after culture. This study investigated the ability of B. thuringiensis GS1 to induce resistance to Rhizoctonia solani KACC 40111 (RS) in cucumber plants. Chitinase activity was greatest in RS-treated plants at 4 days. ${\beta}$-1,3-Glucanase activity was highest in GS1-treated plants at 5 days. Guaiacol peroxidase (GPOD) activity increased continuously in all treated plants for 5 days. Ascorbate peroxidase (APX) activity in RS-treated plants was increased 1.5-fold compared with the control at 4 days. Polyphenol oxidase (PPO) activity in RS-treated plants was increased 1.5-fold compared with the control at 3 days. At 5 days after treatment, activity staining revealed three bands with chitinase activity (Ch1, Ch2, and Ch3) on SDS-PAGE of cucumber plants treated with GS1+RS, whereas only one band was observed for RS-treated plants (Ch2). One GPOD isozyme (Gp1) was also observed in response to treatment with RS and GS1+RS at 4 days. One APX band (Ap2) was present on the native-PAGE gel of the control, and GS1- and GS1+RS-treated plants at 1 day. PPO bands (Po1 and Po2) from RS- and GS1+RS-treated plants were stronger than in the control and GS1-treated plants upon native-PAGE at 5 days. Taken together, these results indicate that the induction of PR proteins and defense-related enzymes by B. thuringiensis GS1 might have suppressed the damping-off caused by R. solani KACC 40111 in cucumber plants.

• Journal of Microbiology and Biotechnology
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• 제24권12호
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• pp.1699-1706
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• 2014

A lichenase gene (mt-lic) was identified for the first time through function-based screening of a soil metagenomic library. Its deduced amino acid sequence exhibited a high degree of homology with endo-${\beta}$-1,3-1,4-glucanase (having both lichenase and chitosanase activities), encoded by the bgc gene of Bacillus circulans WL-12. The recombinant lichenase overexpressed and purified from Escherichia coli was able to efficiently hydrolyze both barley ${\beta}$-glucan and lichenan. The enzyme showed maximal activity at a pH of 6.0 at $50^{\circ}C$, with Azo-barley-glucan as the substrate. The metal ions $Mn^{2+}$, $Mg^{2+}$, $Ca^{2+}$, and $Fe^{2+}$ enhanced the enzymatic activity, whereas the $Cu^{2+}$ and $Zn^{2+}$ ions inhibited the enzymatic activity. The $K_m$ and $V_{max}$ values of the purified lichenase were determined to be 0.45 mg/ml and 24.83 U/min/mg of protein, respectively.

• 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.

• Journal of Microbiology and Biotechnology
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• 제27권2호
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• pp.271-276
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• 2017

A highly thermostable ${\beta}-(1-4)-glucanase$ (NA23_08975) gene (fig) from Fervidobacterium islandicum AW-1, a native-feather degrading thermophilic eubacterium, was cloned and expressed in Escherichia coli. The recombinant FiG (rFiG) protein showed strong activity toward ${\beta}-{\small{D}}-glucan$ from barley (367.0 IU/mg), galactomannan (174.0 IU/mg), and 4-nitrophenyl-cellobioside (66.1 IU/mg), but relatively weak activity was observed with hydroxyethyl cellulose (5.3 IU/mg), carboxymethyl cellulose (2.4 IU/mg), and xylan from oat spelt (1.4 IU/mg). rFiG exhibited optimal activity at $90^{\circ}C$ and pH 5.0. In addition, this enzyme was extremely thermostable, showing a half-life of 113 h at $85^{\circ}C$. These results indicate that rFiG could be used for hydrolysis of cellulosic and hemicellulosic biomass substrates for biofuel production.