• Journal of Life Science
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• v.25 no.12
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• pp.1408-1414
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• 2015

In this study, to retain a stable bacterial inoculant, Bacillus strains showing antifungal activity were screened. The improved production, antifungal mechanism, and stability of the antifungal metabolite by a selected strain, AF4, a potent antagonist against phytopathogenic Botrytis cinerea, were also investigated. The AF4 strain was isolated from rhizospheric soil of hot pepper and identified as Bacillus subtilis by phenotypic characters and 16S rRNA gene analysis. Strain AF4 did not produce antifungal activity in the absence of a nitrogen source and produced antifungal activity at a broad range of temperatures (25-40℃) and pH (7-10). Optimal carbon and nitrogen sources for the production of antifungal activity were glycerol and casein, respectively. Under improved conditions, the maximum antifungal activity was 140±3 AU/ml, which was higher than in the basal medium. Photomicrographs of strain AF4-treated B. cinerea showed morphological abnormalities of fungal mycelia, demonstrating the role of the antifungal metabolite. The B. subtilis AF4 culture exhibited broad antifungal activity against several phytopathogenic fungi. The antifungal activity was heat-, pH-, solvent-, and protease-stable, indicating its nonproteinous nature. These results suggest that B. subtilis AF4 is a potential candidate for the control of phytopathogenic fungi-derived plant diseases.

• Food Science and Biotechnology
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• v.17 no.3
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• pp.679-682
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• 2008

A lactic acid bacterium producing an antimicrobial substance against Escherichia coli O157:H7 was isolated from raw milk and identified as Lactobacillus amylovorus ME-1. In addition to E. coli O157 :H7, the antimicrobial substance also inhibited the growth of Bacillus cereus, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus pyrogenes, and Yersinia enterocolitica. The antimicrobial substance was stable at pH 2-12 and $121^{\circ}C$ for 15 min and insensitive to proteinase K, protease, amylase, and catalase. Purification of the antimicrobial substance was conducted through methanol and acetonitrile/ethylacetate extraction, ultrafiltration with a 500 Da cutoff, thin layer chromatography (TLC) with silicagel 60, and high performance liquid chromatography (HPLC) with a $C_{18}$ reverse phase column. The ${\lambda}_{max}$ of the purified antimicrobial substance was determined as 192 nm by ultra violet (UV) scanning, while the molecular weight was estimated as 453 Da based on the mass spectrum. Accordingly, the current results suggest that the antimicrobial substance from the L. amylovorus ME-1 was not a bacteriocin, but rather a new non-proteinaceous substance distinct from acidophilin, acidolin, diacetyl, and reuterin.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.513-516
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• 2001

This study was to develop of efficient microbial agent for malodor removal. Total ten strains of beneficial bacteria Bacillus sp., Pseudomonas sp., and photosynthetic bacteria were isolated and identified on the basis of their morphological and biochemical characteristics. The enzyme activities such as amylase, protease, lipase and cellulase of bacteria cells were measured. Furthennore, effective formulation procedure 、 ,vas developed with nutrient additive, stabilizing agent and mineral materix. For preparation of microbial agent, developing of formulation technique was very helpful for incresing the cell survival rate.

• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.363-367
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• 2006

Food wastes were decomposed into the Mugri (Isung Engineering, Korea), a food waste reduction machine, with adding sawdust of cryptomeria. Degradation effects were better when the machine worked at over 45$^{\circ}C$ than those at the lower temperature. Thermophilic bacteria were isolated from cryptomeria sawdust and the food waste products degraded by the machine. The isolates from cryptomeria sawdust were classified into 3 genera (Acinetobacter baumannii, Enterobacter sp. and Erwinia cypripedii) and almost all the isolates from the degraded products were partially identified as Bacillus sp. by 16S rDNA sequence analysis. The isolated thermophilic bacteria showed degradative enzyme activities. In the case of addition of the 30 thermophilic bacteria into the machine, degradation rate of food wastes was almost twice as high with increasing process temperature up to 6$^{\circ}C$.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.6 no.4
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• pp.342-348
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• 2005

In this study, novel strains showing better protein degradation activity were isolated for the production of effective compost from garbages. Well growing bacteria with clear zone on the skim milk agar media were isolated . The strain was identified as Bacillus subtilis PNV-1 through various biochemical tests, Bergy's manual of determinative bacteriology and 165 rDNA partial sequence. The extracellular protease of the strain PNV-1 has its activity at broad pH and the optimal temperature was $50^{\circ}C$. Also, the strain PNV-1 was highly tolerant to high concentration of salt, red/black pepper and mustard.

• Journal of Sericultural and Entomological Science
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• v.34 no.2
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• pp.41-51
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• 1992

In this thesis, both soap and enzymatic degumming method were adopted and the optimum degumming conditions were obtained. Difference between the two degumming methods in silk fiber state was investigated and analyzed on the basis of the results of physical testings, polarizing microscopy, scanning electron microscopy, viscosity measurement, (${\alpha}$＋$\varepsilon$) amino group contents measurement, birefringence measurement, amino acid analysis, thermal analysis, infrared spectroscopy and x-ray diffraction analysis. The results obtained were summarized as follows; Physical test results of the degummed silk fiber showed that the tenacity and the elongation of enzymatic degummed silk fiber were lower than those of soap degummed fiber. But SEM observation and amino acid analysis showed almost the same tendency in the two degumming methods. The viscosity of enzymatic degummed silk fiber was lower than that of soap degummed fiber, but (${\alpha}$＋$\varepsilon$) amino group contents was higher in the enzymatic degummed fiber. It can be suggested that the enzymatic degummed silk fibroin was more degraded than the soap degummed fibroin. The birefringence, endothermic temperature of DSC spectrum, IR crystallinity and X-ray lateral order factor of enzymatic degummed silk fiber were higher than those of soap degummed fiber. It seems that the enzymatic degummed silk fiber has the higher crystallinity than that of soap degummed one according to the above results. However, it can be inferred that these differences between soap and enzymatic degummed fiber would be lessened if pretreatment and aftertreatment were included in the enzymatic degumming process.

• KSBB Journal
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• v.11 no.1
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• pp.8-14
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• 1996

Bacillus subtillis strains with transition state regulator mutations and a spore mutation were developed for the overexpression of apsE and for the enhancement of expression level. Among the many regulator genes, degU and hpr were chosen as a representative positive and negative regulator for the aprE, respectively. Spo II G was used for the construction of asporogeneous strains. All the mutants were constructed from two protease-deleted strain DB104 and the apsE gene was transformed with an integration vector pMK101. DB104(deg$U^h$(32) $his^+$)::pMK101(Cm) and DB104($\Delta$her(Em))::pMKl01(Cm) show 7-fold and about 2-fold increase in aprE expression level, respectively. But the effect of transition state regulator mutation on the aprE expression was diminished when the integrated aprE gene was amplified by the high concentration of chloramphenicol, i. e. 30 $\mu\textrm{g}$/ml. DB104($\Delta$spoIIG(Pm) degUh(32) his+)::pMK101(Cm) and DB104($\Delta$spoIIG(Pm) $\Delta$hpr(Em))::pMK101 double mutant show 10-fold and 3-fold increase in aprE expression level, respectively. The results suggest that sporulation mutation and transition state regulator mutation have independent and additive effect on the aprE expression, and the same gene dosage effect on the transition state regulator mutation was also identified.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.665-680
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• 1996

Bovine Pneumonic Pasteurellosis는 수송열(輸送熱)로 일반적으로 알려져 있는 질병으로서, 여러가지 요인의 복합적(複合的)인 작용에 의해 발병하는 것으로 알려져 있으나, Pasteurella haemolytica A1이 가장 주요(主要)한 인자(因子)로 밝혀져 있다. P haemolytica A1은 leukotoxin(LKT), lipopolysaccharide(LPS), capsular polysaccharide 등 여러가지의 병원성인자(病原性因子)을 생성한다. 이들 인자중 LKT가 가장 중요한 병원성인자로 밝혀져 있다. 이에 본 실험은 P haemolytical A1의 LKT 유전자를 Bacillus subtilis에서 발현(發現)시킴으로서 LPS에 오염(汚染)되지 않은 LKT을 대량으로 생산할 목적으로 실시되었다. 실험의 첫 단계(段階)로서 pLKT52 plasmid을 Sau3 A1의 제한효소을 이용하여 부분소화(部分消化)시킨 후 이 부분 소화(消化)된 유전자들로부터 3~5kb 크기의 유전자들을 순수분리하여 pUC18와 결합시킨 후 E coli NM522에 형질전환(形質轉換)시켰다. 이때 형질전환된 균주들은 LKT에 대한 단크론 항체인 MAb601을 이용하여 colony blot 법에 의해서 LKT 유전자 보유 및 발현여부(發現與否)을 조사하였다. 이들 양성 clone들은 제한효소분석(制限酵素分析), 염기서열분석(鹽基序列分析) 및 Western blot 등에 의해서 재확인(再確認)하였다. 총 9개의 양성 clone중 위의 방법에 의해서 한 clone을 선택(選擇)하여 lktCA insert를 재분리하여 shuttle vector에 subcloning 하였다. Subcloning된 LKT 유전자들은 shuttle vector의 종류(種類)(pHPS9, p602/20, pHPS9-Sac)와 각기(各其) 다른 종류(種類)의 B subtilis(spoO12A, BR121, WB3O, Raj1105) 숙주내(宿主內)에서 발현정도를 Western blot 법에 의해서 비교(比較)하였다. 이때 최적발현조건(最適發現條件)은 p602/20와 pBL1의 dual plasmid system을 이용하여 B subtilis spoO12A에서 2시간동안 IPTG로 발현을 유도(誘導)하는 것이었다. B subtilis에서 발현된 LKT을 visual 법과 neutral red uptake 법을 이용하여 소 폐포(肺胞) 대식구(大食求)에 대한 biological activity를 확인하였다. 발현된 LKT에 대한 LPS 오염은 LKT을 SDS-PAGE 후 silver stain에 의해서 확인하였다. 본 실험을 통해서 볼 때에 lktCA 유전자를 보유(保有)하고 있는 p602/20는 B subtilis에서 매우 불안정(不安定)하였고, 발현된 LKT는 세균자체(細菌自體)에서 생성되는 protease들에 의해서 파괴(破壞)됨으로서 농도(濃度)가 매우 낮았다. 이러한 문제점들은 다음 단계(段階)의 실험에서 해결되어야할 문제들이다.

• Korean Journal of Food Science and Technology
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• v.5 no.3
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• pp.163-168
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• 1973

In order to study on the effects of Bacillus natto on the soybean paste used natto, there were determind the contents of total nitrogen, amino acid, ammonia-nitrogen, total acid, reducing sugar and proteolytic activity etc. The results are summerized as follows; 1) Fermentation of natto paste were faster than koji paste and the fermentation period were sufficient in thirteen days. 2) The chemical components, color, taste and flavor of natto paste (added 15% natto) were excellent than the other soybean paste. 3) The contents of ammonia-nitrogen and amino acid of soybean paste were gradually increased during thirteen days in the fermentation, while total nitrogen contents were decreased. 4) After thirteen days of the fermentation, the contents of amino acid were decreased. 5) The contents of amino acid of natto paste were much more those of koji paste. 6) Total acid and reducing sugar contents indicated a rapid increased from nine to fifteen days during the fermentation, and a slower and decrease after fifteen days. 7) Total acid and reducing sugar contents of natto paste were somewhat higher than those of koji paste. 8) No significant difference was observed in the contents of reducing sugar and total acid according to the amount of natto added. 9) The contents of reducing sugar and amino acid were proportional to the proteolytic activity. 10) A flood soybean paste may be produced by adding 15% natto to the raw material in thirteen days of the fermentation.

• Korean Journal of Food Science and Technology
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• v.33 no.3
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• pp.338-344
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• 2001

The effect of gamma-irradiation on the microbiological and general qualities of Kanjang (fermented soy sauce, Korean type) and Shoyu (fermented soy sauce, Japanese type) was studied. Samples were prepared, irradiated at 5, 10 and 20 kGy, and then stored at $25^{\circ}C$ for 18 weeks. The results showed that the Bacillus spp. was inactivated by 3 log cycles with the irradiation dose of 10 kGy and the number of Bacillus cells that survived from 10 to 20 kGy of gamma irradiation was decreased gradually during storage. Yeasts and Lactobacillus were nearly eliminated at 5 to 10 kGy of irradiation. The $D_{10}$ values of Bacillus, yeast and Lactobacillus in Shoyu were 2.67 kGy, 0.81 kGy and 1.30 kGy, in Kanjang were 2.75 kGy, 0.99 kGy and 1.47 kGy, respectively. The general quality of gamma irradiated Shoyu and Kanjang, such as total nitrogen, amino nitrogen, protease activity and pH were more stable than that of control during storage periods. Decolorization was observed just after irradiation, but the color was recovered during storage. The sensory evaluations showed that irradiated samples were more acceptable. Therefore, it was considered that gamma irradiation was effective for maintaining better quality of Shoyu and Kanjang during storage.