The purpose of this study was to evaluate the appearance, physicochemical, physical, and fermentation properties of the fermented soybean produced by manufacturing with inoculation the different types of microbial strains. The strains were inoculated by the NSI (natural strains inoculation), and the SSI (selective strain inoculation) were treatments. The appearance showed differences in color, viscous substance, and hardness depending on strains inoculation and fermentation duration. The pH, and total acidity were 6.40~7.26%, and 0.10~0.39% respectively with differences depending on the samples. The moisture content as the fermentation duration increased, the NSI (56.03~57.66%) decreased and the SSI (56.71~58.63%) increased. The physical characteristics of the hardness increased as the fermentation duration increased for the NSI and the SSI decreased. The color values for the L, a, and b values were 47.64~58.56, 7.15~9.08, and 12.41~17.30, respectively. The α-amylase and protease activities of the SSI were the highest among all treatments. The total viable cell counts of the fermented soybean products by strains were 5.02 to 9.77 log CFU/g, and SSI (fermentation, 48 hours) was the highest. The amino-type nitrogen contents of all samples were 301.62~746.97 mg% and the SSI showed the highest content. The amino acid had the highest glutamic acid content.

The occurrence of allergic disease has increased harmfully in the last few decades. Atopic dermatitis (AD) is an allergic inflammation disorder characterized by itchy, red, swollen, cracked skin. Although the pathogenesis of AD is not fully understood, it is assumed that deregulation of T-helper 1 (Th1) and T-helper 2 (Th2) immune responses, a predominance of allergen-specific IgE, and interrupted epidermal barrier function are keys to the pathogenic mechanism. Activated T helper 2 (Th2) immune function is hallmark of various allergic diseases. Oxidative stress implicated in cutaneous damage in various inflammatory skin diseases. We investigated the effect of fermented soybean (SCGB1) on the improvement of AD. Soybean fermentation was carried out using B. amyloliquefaciense SCGB1 (KCCM11964P), which is known to produce of natural antibiotics. And then, we experiment of SCGB1 and soybean powder (NC) in DNCB-induced AD model. Mice were respectively oral administration of variety dose for 14 days. As a results, it was confirmed that serum Immunoglobulin E (IgE) expression was dose-dependently decreased in SCGB1 and NC compared to negative control, and it was reduced the skin pruritus inducing factor that Interleukin-31 (IL-31) mRNA level. In addition, the inflammatory cells were infiltration in skin for histological analysis. As a result, it reduced that epidermal hyperplasia, cancellation and aveolarization compared to negative control. These results suggest that SCGB1 may be effective for prevention and treatment of AD.

The button mushroom, Agaricus bisporus, is one of the major economical crops cultivated in Korea. This mushroom showed the 5th production to 11,493 M/T in 2014. This study examined changes in decomposition-related enzyme activity secreted by the microorganisms isolated from button mushroom composts. Among the isolated microorganisms, bacterial strains CY-36 were the most dominant and had the highest enzyme activities. 16S rDNA sequencing analysis showed 99% similarity of CY-36 to Bacillus amyloliquefaciens. To define the optimum conditions for the mass production of CY-36, we have investigated optimum culture conditions and effects of various nutrient source on the bacterial growth. The optimum initial pH and temperature were determined as pH 5.0~8.0 and 30°C, respectively. The optimal culture medium for the growth was determined as follows: 2.0% dextrin, 1.0% yest extract, 0.5% NaNO3, 3 mM MgSO4, and 0.6% valine. Enzymatic activities of CMCase and PGase secreted by CY-36 were the most prominent and CMCase activity were optimal at 40°C and pH 7.0, and PGase activity were 50°C and pH 6.0. The activity of CMCase was rapidly decreased at more than 4°C and pH 5.0, and the activity of PGase was rapidly decreased at more than 30°C but didn’t affect as pH change. Accordingly, Bacillus amyloliquefaciens CY-36 was potentially useful on the fermentation stimulation and softening of rice straw in the process of fermentation for mushroom cultivation.

고추를 비롯한 채소작물에 심각한 피해를 야기하는 흰비단병의 생물학적 방제제를 개발하기 위하여 시설재배 토양에서 분리한 세균 중에서 흰비단병균인 Sclerotium rolfsii의 균사생장을 강하게 억제하는 균주를 선발하였다. 선발된 균주는 형태적, 생리적 분석과 16S rRNA 분석결과를 바탕으로 Bacillus amyloliquefaciens로 동정하였고 최종 선발균주를 Bacillus amyloliquefaciens KBC1009로 명명하였 다. Bacillus amyloliquefaciens KBC1009의 배양특성을 조사한 결과 배양온도 30℃, pH 7의 조건에서 72시간동안 배양했을 때 배양 효율이 가장 우수하였으며, 배양에 적정한 영양원은 탄소원 glucose, 질 소원 yeast extract를 사용했을 때 가장 높은 균수를 나타내었다. 포장에서 고추 흰비단병에 대한 Bacillus amyloliquefaciens KBC1009의 방제효과를 검정하기 위해 흰비단병원균을 토양에 접종하고 B. amyloliquefaciens KBC1009 배양액(5×108 cfu/ml)을 500배로 희석하여 10일 간격으로 3회 관주 처리한 결과 무처리대비 66.7%의 방제효과를 나타내었다.

The mannanase-producing bacteria, designated YJ17, was isolated from spent mushroom (Flammulina velutipes) substrates. The isolate YJ17 was a facultative anaerobic and was grown at temperatures ranging from 20oC to 50oC with an optimal temperature of 40oC. The DNA G+C content of the YJ17 was 44 mol%. The major fatty acids were anteiso-15:0 (38.9%), 17:0 (7.6%), and iso-15:0 (36.5%). The 16S rRNA gene sequence similarity between the isolate YJ17 and other Bacillus strains was from 98% to 99%. In the phylogenetic analysis based on these sequences, the isolate YJ17 and Bacillus amyloliquefaciens clustered within a group together and separated from other species of Bacillus. Based on the physiological and molecular properties, the isolate YJ17 was classified within the genus Bacillus as B. amyloliquefaciens YJ17. The optimal pH and temperature for mannanase activity of B. amyloliquefaciens YJ17 were pH 7.0 and 50oC, respectively.

Spent mushroom substrate (SMS) is a by-product remaining after a crop of mushrooms. The fresh SMS was sample within 24 hours of removal from the production facility in DOJUN farm located in Jinju. About 11 bacterial species were isolated from fresh SMS on TSA medium. Among of them, one isolate, designated YJ02, showed the antifungal activity against Aspergillus flavus and Aspergillus ochraceous producing mycotoxin on PDA medium, potentially. The strain YJ02 was produced cellulase, xylanase, mannanase as hydrolase. The strain YJ02 was identified as members of the genus Bacillus by biochemical characteristics using Bacillus ID kit and VITEK 2 system. Comparative 16S rDNA gene sequence analysis showed that strain YJ02 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus amyloliquefaciens with 16S rDNA gene sequence similarity of 98.7%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, strain YJ02 was classified within the genus Bacillus, for which the name Bacillus amyloliquefaciens YJ02 is proposed.

Bacillus subtilis and Bacillus amyloliquefaciens are closely related species that share a similar genomic background, and are both known to secrete large amounts of proteins directly into a medium. The extracellular proteomes of two strains of Bacillus subtilis and two strains of Bacillus amyloliquefaciens were compared by 2-D gel electrophoresis during the late exponential growth phase. The relative abundance of some minor protein spots varied among the four strains of Bacillus. Over 123 spots of extracellular proteins were visualized on the gel for B. subtilis CH 97, 68 spots for B. subtilis 3-5, 230 spots for B. amyloliquefaciens CH 51, and 60 spotsfor B. amyloliquefaciens 86-1. 2D gel electrophoresis images of the four Bacillus strains showed significantly different protein profiles. Consistent with the 2D gel electrophoretic analysis, most of the B. subtilis proteins differed from the proteases secreted by the B. amyloliquefaciensstrains. Among the proteins identified from B. subtilis, approximately 50% were cytoplasmic and 30% were canonically extracellular proteins. The secreted protein profiles for B. subtilis CH 97 and B. subtilis 3-5 were quite different, as were the profiles for B. amyloliquefaciens CH 51 and 86-1. The four proteomes also differed in the major protein composition. The B. subtilis CH 97 and B. amyloliquefaciens CH 51 proteomes both contained large amounts of secreted hydrolytic enzymes. Among the four strains, B. subtilis 3-5 secreted the least number of proteins. Therefore, even closely related bacteria in terms of genomic sequences can still have significant differences in their physiology and proteome layout.

버섯 생산 후 발생되는 부산물인 새송이버섯 수확 후 배지로부터 surfactin을 생성하는 4종의 균주를 분리하였으며 이 중 곰팡이 독소를 생성하는 A. flavus와 A. ochraceous 에 대한 항균활성이 우수한 균주를 최종 선발하여 YJ07로 명명하였다. Bacillus ID kit와 VITEK 2 system를 이용하여 분리균 YJ07의 생리적·생화학적 특성을 조사한 결과 분리균은 B. subtilis, B. amyloliquefaciens와 유사한 생리적·생화학적 특성을 나타내었으며 16S rDNA 염기서열 분석을 통한 계통학적 유연관계에서도 B. amyloliquefaciens와 99.5%의 상동성을 나타내었다. 이와 같은 결과를 종합하여 분리균 YJ07은 B. amyloliquefaciens YJ07로 동정되었으며 분리균 YJ07이 생성하는 항균물질은 TLC와 HPLC 분석에서 reference 물질로 사용한 srfactin과 유사한 특성을 나타내었다.

Spent mushroom substrate(SMS) is a by-product remaining after a crop of mushrooms. About 9 thermophilic strains were isolated from SMS(Flammulina velvtipes). Among of them, one isolate, designated UJ09, showed the antifungal activity against Aspergillus flavus and Aspergillus ochraceous producing mycotoxin on PDA medium, potentially. The strain UJ09 was produced cellulase and xylanase as hydrolase. The strain UJ21 was identified as members of the genus Bacillus by biochemical characteristics using Bacillus ID kit and VITEK 2 system. Comparative 16S rDNA gene sequence analysis showed that strain UJ09 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus amyloliquefaciens with 16S rDNA gene sequence similarity of 99.2%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, strain UJ09 was classified within the genus Bacillus, for which the name Bacillus amyloliquefaciens UJ09 is proposed.

A bacterium was isolated from feces of Allomyrina dichotoma larva that feeds on well-rotted sawdust of woods. The isolate was identified as a Gram-positive and spore-forming stain. We named the isolate as Bacillus amyloliquefaciens on the basis of morphological and biochemical properties as well as 16S rRNA gene sequences. The culture of bacteria beneficially increased root and shoot growth of tomato, pepper and cucumber plants compared to the distilled water control. In addition, the bacterial culture strongly inhibited the mycelial growth of several fungal phytopathogens. The drop collapse assay with this culture showed a surfactant activity that is a major indicator for the selection of biocontrol agent. Also, a bacterium has ability of wastewater treatment. These data demonstrate the potential application of B. amyloliquefaciens as a biocontrol agent.

Bacillus amyloliquefaciens GR4-5 was isolated from the rhizosphere soil of Korean ginseng and displayed broad-spectrum suppression of ginseng root rot pathogens. The survivability of B. amyloliquefaciens GR4-5 in soil was investigated under three different conditions; indoor, outdoor ─ of which soil was put in 14 mL tube after treatment ─ and field environments. Soil samples were collected over a fourweek period from three experimental designs, and assessed for 16S rRNA gene copy number by quantitative polymerase chain reaction (qPCR). In outdoor condition, the 16S rRNA gene copy number of Bacillus spp. was 8.35 log copies g soil-1 immediately after the GR4-5 treatment. Two weeks later, the 16S rRNA gene copy number of Bacillus spp. (6.70 log copies g soil-1) was similar to that of the control (6.38 log copies g soil-1). In indoor condition, the 16S rRNA gene copy number of Bacillus spp. maintained in a certain level for a longer period than those in outdoor and field. The 16S rRNA gene copy number of Bacillus spp. in field experiment was reduced faster than that of outdoor condition. Our results show that B. amyloliquefaciens GR4-5 can survive in bulk soil for 1 week, indicating its potential use as a biocontrol agent following 7 day application intervals. This study presents that outdoor microcosm system design could be a useful method to assess easily the survivability of beneficial microorganisms.

본 연구에서는 새로운 프로바이오틱스를 선발하고자 protease를 생산하는 GRAS균주를 된장 및 청국장으로부터 분리하고 그 특성을 조사하였다. 분리한 균주를 분리 동정 한 결과 각각B. amyloliquefaciens CDD5, B. amyloliquefaciens CPD4 및B. amyloliquefaciens CGD3로 명명하였다. 분리균 주의 생육은 12시간 배양 시, 7.13~7.32 log CFU/mL로 최대 생육도를 보였으며 24시간까지 유지되다가 감소하는 경향 을 나타내었고, 그 중B. amyloliquefaciens CGD3의 protease 활성이9.21 U/mL로서가장높게나타났다. B. amyloliquefaciens 이 생산하는 protease 활성은 pH 7.0~10.0, 온도는 50℃에서 최적활성을 나타내었으며, casein에서 protease활성이 가장 우수하게 나타났다. pH, NaCl, glucose 농도를 달리한 배양 조건에 따른 B. amyloliquefaciens의 생육특성으로는 pH 5.0~10.0의 넓은 범위에서 생육하였고, NaCl은 12% 농도까 지 증식하였으며 glucose 5%까지는 생육하나 10% 농도부 터는 현저하게 생육이 낮아짐을 확인하였다. 또한 혈당저 하를 유도하는 α-glucosidase의 저해활성을 측정한 결과, B. amyloliquefaciens CPD4 및B. amyloliquefaciens CGD3에 서는 각각 96.65% 및 97.85%의 저해율을 나타내었다. 이러 한 결과는 특히 protease활성이 가장 우수하고 α-glucosidase 또한 높은 저해활성을 가진 B. amyloliquefaciens CGD3 균 주의 생육특성을 고려하여 단백질 분해 분야의 프로바이오 틱 적용 및 당뇨예방용 기능성식품 소재로의 가능성이 기대 된다.