• KSBB Journal
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• 제15권6호
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• pp.584-588
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• 2000

The optimum conditions for the production of an antifungal antibiotic from Bacillus sp. YJ-63 were investigated. The oprimumized medium consisted of 1.5% soluble starch, 1% tryptone and 0.5% yeast extract, and temperature and initial medium pH for production were optimal at 35$^{\circ}C$ and pH 6.0, respectively. Production yield was significantly improved by shaking culture using 50 ml medium in 500 ml flasks. Under these conditions, the production of the antifungal antibiotic was growth-dependent, from 35hrs into cultivation to the stationary phase and endospore formation.

• Journal of Applied Biological Chemistry
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• 제57권2호
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• pp.159-160
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• 2014

An antifungal antibiotic was extracted three times using n-butanol from the culture broth of Bacillus sp. Bioassayguided column chromatography with silica gel and Sephadex LH-20 yielded 62 mg of the original active compound from 1 L of culture broth. The minimal inhibitory concentration values were 25 and $50{\mu}g/ml$ against Pyricularia oryzae and Pellicularia filamentosa, respectively. Based on results obtained from the analysis of the structure of the antibiotic using MS, NMR, and IR spectroscopy, the antifungal antibiotic was shown to consist of only six of fructose.

• 대한치과보철학회지
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• 제32권2호
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• pp.268-280
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• 1994

Tissue conditioners have been used for treatment of denture stomatitis caused by wearing of dentures. Early studies pointed out Candide albicans (C. albicans) as main etiologic factor, and antifugal agents were added for control of the species. But there is a little information about broad comparison on the effect of tissue conditioners and antifungal agents added. The purpose of the present study was to compare the inhibiting effect of four tissue conditioners and one temporary soft liner on the growth of C. albicans for treatment of denture stomatitis using gel diffusion method by measuring diameter of the zone of growth inhibition. Three antifungal agents were added to each material for evaluation of the effect of added agents. Finally, observation was made to evaluate the effect of the loss of antifungal elements by aging of the specimen. The results of this study were obtained as follows : 1. Tempo had remarkable antifungal effect showing the zone of growth inhibition as 2.35 mm at 1st day, and was most effective on End: 4th and 7th day from incubation (p<0.05). But Coecomfort, Dura conditioner, Visco-gel, Coe-soft had little antifungal effect from the 1st day of incubation. 2. Nystatin was most effective showing 9.60-12.04 mm of zone of inhibition at the 1st day from incubation. The antifungal properties were reduced to amphotericin B, chlorhexidine and materials without agent (p<0.05), and the effect was diminished by time. 3. As pretreatment with amphotericin B, nystatin, chlorhexidine, Tempo was very effective at the 1st day from incubation showing zone of inhibition as 3.65, 12.04, 4.78 mm with addition of each agent. Dura conditioner had strongest antifungal effect at the next day as 2.86, 5.33, 1.29 mm of zone of inhibition, and yielding results of Coe-comfort, Tempo, Coe-soft was shown at 4th and 7th day from incubation (p<0.05). Taken all together, tissue conditioners have little antifungal effect except Tempo. Formation of the zone of growth inhibition was due to agents amphotericin B, nystatin, chlorhexidine and nystatin was most effective. Conclusively, it is advisable to select material which is effective on the growth of C. albican and consider addition of antifungal agents for treatment of denture stomatitis.

• 생명과학회지
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• 제20권9호
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• pp.1339-1344
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• 2010

환경친화적인 항균발효퇴비를 생산할 목적으로 식물병원성 균주에 대하여 항균활성이 우수한 B. licheniformis KJ-9를 톱밥우분에 접종하여 고품질의 항균발효퇴비(B. licheniformis KJ 9 fermented compost, BLC)의 개발을 행하였다. B. licheniformis KJ-9가 생산한 물질의 온도 안정성은 $100^{\circ}C$, 10분간 열처리하여도 약 60% 이상의 항균활성을 유지하였으며, pH 안정성은 산성보다는 pH 7.0 이상의 중성 및 알칼리성 영역에서 높은 항균활성을 유지하였다. 퇴비를 제조하는 과정에서 BLC의 경우는 우분 냄새가 확실히 줄어들었으며 또한 퇴비의 발효시간도 3일 정도 단축되었다. 고추의 pot실험 결과 BLC를 첨가한 시험구 이외의 다른 시험구에서는 고추 잎이 마르고 약간의 황색반점이 발생하였다. 그리고 BLC를 이용한 시험구는 밭 흙만을 사용한 시험구 및 밭 흙에 기존의 판매퇴비(commercial available compost, CC)를 혼합한 시험구보다 잎의 수가 많고(1.5~2배), 줄기 및 뿌리의 성장이 빠르며, 줄기가 굵은(1.5~3배) 것으로 나타났다. 마늘 및 들깨를 재배한 현장적용 실험결과 두 시험구 모두 자연발생적인 병충해의 피해는 없었지만 BLC를 사용한 밭에서 마늘 및 들깨의 생육이 현저하게 빠른 것을 관찰할 수 있었다.

• 생명과학회지
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• 제19권11호
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• pp.1672-1678
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• 2009

B. amyloliquefaciens IUB158-03균주에서 정제된 항진균물질은 극성인 용매에 잘 용해되고, pH 6.0~10.0와 $-70{\sim}121^{\circ}C$에서와 같이 넓은 범위의 온도 및 pH에서 안정성을 보였다. 항진균물질의 FAB-MS, UV 흡수 스펙트럼, 아미노산 조성 등을 분석한 결과 분자량은 1,042 이었고, TLC를 이용하여 분석한 결과 ninhydrin solution에서 보라색으로 발색되었다. UV 스펙트럼은 220 nm, 277 nm에서 ${\lambda}max$를 보였으며, $Asn_3$, $Gln_2$, $Ser_1$ $Gly_1$, $Tyr_1$의 아미노산 조성을 갖는 것으로 나타났다. 그리고 $^1H$-NMR spectrum, $^1H$-COSY, HMQC 을 분석한 결과 iturin A계에 속하는 물질로 확인되었다. NIH3T3 섬유아세포에 대해 항진균물질이 세포독성을 나타내지 않는 것은 물론 마우스에 항진균물질을 경구투여하여 장기 내의 변화와 백혈구 수, 생체내의 생리적인 기능면에서 정상 마우스와 차이를 보이지 않았으므로 생체독성이 없는 것으로 나타났다. 따라서 본 연구를 통하여 B. amyloliquefaciens IUB158-03에서 분리된 항진균물질이 앞으로 고추탄저병의 생물적 방제제로 이용될 수 있는 잠재성을 갖고 있는 것으로 사료된다.

• 한방안이비인후피부과학회지
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• 제16권1호
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• pp.77-93
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• 2003

This experimental study was performed to investigate the antifungal effects of mixture of Sophorac Subprostratae Radix, Aconti Radix, and Hibisci Syriaci Cortex by the three types of extraction in the dermatophytes such as Trichophyton rubrum, Trichophyton mentagrophytes, and Epidermophyton floccosum. The following results were obtained by using Minimum Inhibition Concentration(MIC) and Inhibition Zone. 1. MIC on Trichophyton mentagrophytes in Sample S-2, Sample S-3, and Sample-Y were 10${\mu}l$, respectively, showing the highest antifungal effect. 2. MIC on Epidermophyton floccosum in Sample S-2, Sample S-3, and Sample-Y were 10㎕ respectively, showing the highest antifungal effect. 3. MIC on Trichophyton rubrum in Sample S-3, Sample-Y were 10${\mu}l$ respectively, showing the highest antifungal effect. 4. The size of inhibition zone on Trichophyton mentagrophytes for Sample S-3 were 22.3mm in 10${\mu}l$, showing the highest antifungal effect. 5. The size of inhibition zone on Epidermophyton floccosum for Sample S-3 was 34.3mm in 10${\mu}l$, showing the highest antifungal effect. 6. The size of inhibition zone on Trichophyton rubrum for Sample-Y was 26.5mm in 10${\mu}l$, showing the highest antifungal effect. Whereas. in extractions, that of Sample S-3 was 12.5mm in 10${\mu}l$, showing the highest antifungal effect. 7. From the above results, Sample S-3 on Trichophyton mentagrophytes, Epidermophyton floccosum and Sample-Y on Trichophyton rubrum showed the highest antifungal effects. This results suggests that vinegar extraction excels other extraction method using water or ethanol to measure the antifungal effect on dermatophytes. In addition, the result shows that the extract of herbal medicines can be used to tinea pedis and if further study is performed, the use of the extract of herbal medicines will be valuable and beneficial in the clinical medicines. [NOTE] · S-S2 : 12-13w/v$\%$ Vinegar extract of mixture of Sophorae Subprostratae Radix, Aconti Radix, and Hibisci Syriaci Cortex. · S-S3 : 18-19w/v$\%$ Vinegar extract of mixture of Sophorae Subprostratae Radix, Aconti Radix, and Hibisci Syriaci Cortex. · S-Y : Miconazole nitrate

• Journal of Microbiology and Biotechnology
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• 제10권6호
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• pp.805-810
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• 2000

An antifungal protein antagonistic to the rice blast fungus, Pyricularia oryzae was purified from Paenibacillus macerans PM-1 by ammonium sulfate fractionation, Q Sepharose Fast Flow column chromatography, Phenyl Sepharose CL-4B column chromatography and Superose 12 gen filtration. An apparent molecular mass of the purified antifungal protein was determined as 8 kDa by SDS-PAGE and 9 kDa by analytical gel filtration, respectively, suggesting that the purified protein is a monomer. The antifungal protein was stable at pH range from 7-12 and up to $100^{\circ}C$. The protein was also stable at 0.1-1% Tween 20 and Triton X-100. The N-terminal amino acid sequence of the antifungal protein was Thr-Glu-Leu-Pro-Leu-Gly-Ile-Val-Met-Asp-Lys-Tyr-Thr-Asp-Ala-Phe-Lys-Phe-Asp-Met-Phe. Comparison of the determined sequence with other peptide and DNA sequences did not reveal homology at all. Therefore, the purified antifungal protein was speculated to be a novel protein. The condidial germination in vitro of P. oryzae KJ301:93-39 by the purified protein ($5.9{\mu} g/ml$) was limited to $9{\pm}3.2%$ only, compared with $69{\pm}2.4%$ of the control. Ungerminated conidia were swollen at basa and mid cell by the purified protein. In vivo bioassay for inhibition of conidial germination of P. oryzae KJ 301, one of the most predominating racesin Korea. the purified protein ($5.9{\mu} g/ml$)strongly inhibited the conidial germination. The conidia, even though germinated, could not develop any further to produce appressoria efficiently.

• Journal of Microbiology and Biotechnology
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• 제16권3호
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• pp.450-456
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• 2006

Pseudomonas fluorescens MC07 is a growth-promoting rhizobacterium that suppresses mycelial growth in fungi such as Rhizoctonia solani, Pythium ultimum, Fusarium oxysporum, and Phytophthora capsici. To determine the role of the bacterium's antifungal activity in disease suppression, we screened 2,500 colonies generated by Tn5lacZ insertions, and isolated a mutant 157 that had lost antifungal activity. The EcoRI fragment carrying Tn5lacZ was cloned into pBluescript II SK(+) and used as a probe to isolate wild-type clones from a genomic library of the parent strain, MC07. Two overlapping cosmid clones, pEH4 and pEH5, that had hybridized with the mutant clone were isolated. pEH4 conferred antifungal activity to the heterologous host P.fluorescens strain 1855.344, whereas pEH5 did not. Through transposon mutagenesis of pEH4 and complementation analyses, we delineated the 14.7-kb DNA region that is responsible for the biosynthesis of an antifungal compound. DNA sequence analysis of the region identified 11 possible open reading frames (ORF), ORF1 through ORF11. A BLAST search of each putative protein implied that the proteins may be involved in an antifungal activity similar to polyketides.

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

• 한국식물병리학회지
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• 제14권3호
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• pp.191-202
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• 1998

About 300 actinomycetes were isolated from two forest and one sea-shore soil and tested for inhibitory effects on mycelial growth of six plant pathogenic fungi Magnaporthe grisea, Alternaria mali, Colletotrichum gloeosporioides, Phytophthora capsici, Fusarium oxysporum f. sp. cucumerinum, and Rhizoctonia solani. Among 300 actinomycetes tested, only 16 actinomycetes showed the antifungal activity against the test fungi. Isolate NH 50 was selected for production and purification of antifungal antibiotic substances. Actinomycete isolate NH 50 displayed the broad antifungal spectra against 11 plant pathogenic fungi. To identify actinomycete isolate NH 50, cultural characteristics on various agar media, diaminopimelic acid type, and morphological characteristics by scanning electron microscopy were examined. As a result, actinomycete isolate NH 50 was classified as a rare actinomycete that had LL-DAP type and did not produce spores. After incubation of isolate NH 50 in yeast extract-malt extract-dextrose broth, antifungal compound NH-B1 that inhibited mycelial growth of some plant pathogenic fungi was purified from the methanol eluates of XAD-16 resins by a series of purification procedures, i.e., silica gel flash chromatography, C18 flash chromatography, Sephadex LH-20 column chromatography, silica gel medium pressure liquid chromatography (MPLC), C18 MPLC, and high pressure liquid chromatography (HPLC). UV spectrum and 1HNMR spectrum of antifungal compound NH-B1 dissolved in methanol were examined. The antibiotic NH-B1 showed the major peaks at 230 and 271.2nm. Based on the data of 1H-NMR spectrum, NH-B1 was confirmed to be an extremely complex polymer of sugars called polysaccharides. The antibiotic NH-B1 showed strong antifungal activity against Alternaria solani and Cercospora kikuchi, but weak activity against M. grisea.