• Journal of Microbiology and Biotechnology
• /
• 제19권10호
• /
• pp.1169-1175
• /
• 2009

An antifungal chitinase, ChiCW, produced by Bacillus cereus 28-9 is effective against conidial germination of Botrytis elliptica, the causal agent of lily leaf blight. ChiCW as a modular enzyme consists of a signal peptide, a catalytic domain, a fibronectin type-III-like domain, and a chitin-binding domain. When two C-terminal domains of ChiCW were truncated, $ChiCW{\Delta}FC$ (lacking the chitin-binding domain and fibronectin type III-like domain) lost its antifungal activity. Since $ChiCW{\Delta}C$ (lacking the chitin-binding domain) could not be expressed in Escherichia coli as $ChiCW{\Delta}FC$ did, a different strategy based on protein engineering technology was designed to investigate the involvement of the chitin-binding domain of ChiCW ($ChBD_{ChiCW}$) in antifungal activity in this study. Because ChiA1 of Bacillus circulans WL-12 is a modular enzyme with a higher hydrolytic activity than ChiCW but not inhibitory to conidial germination of Bo. elliptica and the similar domain composition of ChiA1 and ChiCW, the C-terminal truncated derivatives of ChiA1 were generated and used to construct chimeric chitinases with $ChBD_{ChiCW}$. When the chitin-binding domain of ChiA1 was replaced with $ChBD_{ChiCW}$, the chimeric chitinase named ChiAAAW exhibited both high enzyme activity and antifungal activity. The results indicate that $ChBD_{ChiCW}$ may play an important role in the antifungal activity of ChiCW.

• 식물병연구
• /
• 제16권1호
• /
• pp.35-40
• /
• 2010

식물근권에서 유래된 12개의 근권세균이 Phytophthora citrophthora에 의해 발생되는 감귤 역병에 대해 감귤 열매에서 병진전 억제 효과를 나타내는지 조사하였다. 조사한 근권세균 중 THJ609-3, TRH423-3, BRH433-2, Lysochit 및 KRY505-3 등에 의해 역병이 억제되는 것을 역병균의 in vivo 상처접종을 통하여 밝혀졌다. 근권세균을 P.citrophthora의 균사체와 대치 배양하여 역병균 저지대의 길이를 측정한 결과 역병 진전억제효과를 보였던 5개의 균주에서 모두 항진균활성이 나타났다. 그러나 근권세균의 균사생장억제효과와 역병억제효과 사이에 양의 상관관계는 성립되지는 않았다. 한편, 근권세균 rDNA의 internal transcript spaces(ITS)을 분석을 통해 동정 한 결과 Lysochit과 KRY505-3은 Bacillus cereus로 동정 되었고, BRH4332은 B. circulans로, TRH423-3는 Burkholderia gladiol로 동정되었다. 이 연구는 친환경 농가와 같이 농약사용이 제한된 농장에서 감귤 역병에 대해 생물적 방제를 위한 활성균을 모색하는데 매우 가치가 있을 것으로 생각된다.

• 한국약용작물학회지
• /
• 제24권3호
• /
• pp.191-197
• /
• 2016

Background: This study examined the use of new bio-materials with enhanced value and functionality, which were derived from fermented wild ginseng cultures. Methods and Results: To examine the antioxidant activity associated with biological functions, radical scavenging analyses (2,2-diphenyl-1-picrylhydrazyl, DPPH and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid, ABTS) and superoxide dismutase (SOD)-like activity analyses were conducted. Furthermore, the total phenolic and flavonoid contents of wild ginseng fermented with microorganisms (Leuconostoc mesenteroides, Bacillus circulans, Bacillus licheniformis and B. subtilis subsp. inaquosorum) were evaluated to determine the antioxidant activity increment. Regarding ginseng fermented with B. licheniformis, values of $70.6{\pm}1.4%$, $44.3{\pm}1.7%$, and $88.4{\pm}1.3%$ were measured using DPPH, ABTS, and SOD-like antioxdiant activity analyses, respectively. The total phenolic content in ginseng fermented with B. licheniformis was $184.5{\pm}0.9{\mu}g{\cdot}GAE/m{\ell}$, and the total flavonoid contents was $108.5{\pm}1.8{\mu}g{\cdot}QE/m{\ell}$ in ginseng fermented with L. mesenteroides. Conclusions: Of the four types of lactic acid bacteria examined, the use of B. licheniformis to ferment ginseng resulted in greatest increase in antioxidant activity. Therefore, ginseng fermented by microorganisms might be used to produce functional bio-materials.

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

• 대한방사선기술학회지:방사선기술과학
• /
• 제41권5호
• /
• pp.427-435
• /
• 2018

There was a shortage of research reports on sterilization criterion and contamination of ultrasonic probes. Therefore, in this study, we were going to provide a basic study to measure the level of microbial contamination in ultrasonic probes and to investigate the radiographer's awareness of infection. After the scan, samples were collected from the rubber part of the probe by opening a sterile swab (Transport Medium AM608-1S) for medical bacteria collection with the remaining gel removed with a paper towel. Also, the collected samples of bacteria were grown for seven days and then the laboratory was analyzed. Among the total 29 types of microorganisms, Micrococcus luteus 21(26%), Moraxella species 16(20%), Coagulase negative staphylococcus 8(10%), Bacillus species 5(7%), Bicillus circulans 3(5%), Acinetobacter lwoffii 2(2%), and 1 other Candida parapsilosis (1%) a number of bacteria and fungus, was detected. In a disinfectant experiment using LuciPac Pen on the Lumitester PD-30s, we cultured the rubber part of the probe two to three times to measure the bacteria. Bacteria decreased to 97% with Aquanax (alkaline reduced water 100%), 99% with Klarion wash (0.01% sodium hydroxide), 94% with Klarion disinfection (0.01% nitrous acid water), Sterilization was best with Klarion wash (0.01% sodium hydroxide). Therefore, guidelines for cleaning and disinfection of ultrasonic probes was required, and further development of probe-only disinfectants is required.

• 한국식품과학회지
• /
• 제9권2호
• /
• pp.97-105
• /
• 1977

1) 효모세포벽(酵母細胞壁) 용해활성(溶解活性)이 큰 미생물을 얻기 위하여 baker's yeast-peptone-bouillon agar 평판(平板)배지에서 투명하게 용해된 부위를 형성하는 균주(菌株)를 서울 및 경기지방의 토양 및 하수(下水)시료에서 156개 분리하였고 이들중 활성(活性)이 가장 큰 균주(菌株)로 K-42를 선발하여 Bacillus circulans로 동정(同定)하였다. 2) 균주(菌株) K-42에 의한 효모세포벽 용해효소의 생산을 보면 당류 첨가의 경우 배양 2일째에는 maltose>glucan>xylose>control의 순으로, 3일째에는 lactose>galactose>glucan>control의 순으로 나타났다. 무기 질소원의 경우는 배양2일째 ammonium acetat>sodium nitrate>control의 순으로, 3일째는 ammonium chloride>ammonium oxalate>control의 순으로 나타났으며 milk casein을 제외한 거의 모든 유기질소원은 배양 2일째 활성(活性)의 증가를 보였으나 3일째는 모두 감소하였다. 당류와 질소원의 배합첨가는 상승효과가 없었다. 3) 효소생산에 미치는 당류와 질소원의 첨가 효과는 배양기간 중 pH의 변화와 깊은 관계가 있었는 바 배양중 $pH\;7{\sim}8$을 계속 유지시켜 주면서 당류 또는 질소원을 첨가하면 높은 활성(活性)을 상당기간 계속 유지할 수 있었다. 4) K-42균주(菌株)가 분비(分泌)하는 세포벽(細胞壁) 용해효소의 작용최적(作用最適) 조건은 $pH\;7{\sim}8$, $60^{\circ}C$이었고 열처리(熱處理) 효모세포벽의 용해율은 65%이었다.

• 한국미생물생명공학회:학술대회논문집
• /
• 한국미생물생명공학회 2001년도 Proceedings of 2001 International Symposium
• /
• pp.40-45
• /
• 2001

Raw starch-digesting amylase (BF-2A, M.W. 93, 000 Da) from Bacillus circulans F-2 was converted to two components during digestion with subtilisin. Two components were separated and designated as BF-2A' (63, 000 Da) and BF-2B (30, 000 Da), respectively. BF-2A' exhibited the same hydrolysis curve for soluble starch as the original amylase (BF-2A). Moreover, the catalytic activities of original and modified enzymes were indistinguishable in $K_{m}$, Vmax for, and in their specific activity for soluble starch hydrolysis. However, its adsorbability and digestibility on raw starch was greatly decreased. Furthermore, the enzymatic action pattern on soluble starch was greatly different from that of the BF-2A. A smaller peptide (BF-2B) showed adsorb ability onto raw starch. By these results, it is suggested that the larger peptide (BF-2A') has a region responsible for the expression of the enzyme activity to hydrolyze soluble substrate, and the smaller peptide (BF-2B) plays a role on raw starch adsorption. A similar phenomenon is observed during limited proteinase K, thermolysin, and endopeptidase Glu-C proteolysis of the enzyme. Fragments resulting from proteolysis were characterized by immunoblotting with anti-RSDA. The proteolytic patterns resulting from proteinase K and subtilisin were the same, producing 63- and 30-kDa fragments. Similar patterns were obtained with endopeptidase Glu-C or thermolysin. All proteolytic digests contained a common, major 63-kDa fragment. Inactivation of RSDA activity results from splitting off the C-terminal domain. Hence, it seems probable that the protease sensitive locus is in a hinge region susceptible to cleavage. Extracellular enzymes immunoreactive toward anti-RSDA were detected through whole bacterial cultivation. Proteins of sizes 93-, 75-, 63-, 55-, 38-, and 31-kDa were immunologically identical to RSDA. Of these, the 75-kDa and 63-kDa proteins correspond to the major products of proteolysis with Glu-C and thermolysin. These results postulated that enzyme heterogeneity of the raw starch-hydrolysis system might arise from the endogeneous proteolytic activity of the bacterium. Truncated forms of rsda, in which the gene sequence encoding the conserved domain had been deleted, directed the synthesis of a functional amylase that did not bind to raw starch. This indicates that the conserved region of RSDA constitutes a raw starch-binding domain, which is distinct from the active centre. The possible role of this substrate-binding region is discussed.d.