• Title/Summary/Keyword: Bacillus subtilis

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Neuroprotective effect of fermented ginger extracts by Bacillus subtilis in SH-SY5Y cells (고초균에 의한 생강 발효 추출물의 신경세포 보호 효과)

  • Yang, Hee Sun;Kim, Mi Jin;Kim, Mina;Choe, Jeong-sook
    • Journal of Nutrition and Health
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    • v.54 no.6
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    • pp.618-630
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    • 2021
  • Purpose: The ginger rhizome (Zingiber officinale) is widely cultivated as a spice for its aromatic and pungent components. One of its constituents, 6-hydroxydopamine (6-OHDA) is usually thought to cross the cell membrane through dopamine uptake transporters, and induce inhibition of mitochondrial respiration and the generation of intracellular reactive oxygen species (ROS). This study examines the neuroprotective effect and acetylcholinesterase (AChE) inhibitory activity of fermented ginger extracts (FGEs) on 6-OHDA induced toxicity in SH-SY5Y human neuroblastoma cells. Methods: Ginger was fermented using 2 species of Bacillus subtilis, with or without enzyme pretreatment. Each sample was extracted with 70% ethanol. Neurotoxicity was assessed by applying the EZ-Cytox cell viability assay and by measuring lactic dehydrogenase (LDH) release. Morphological changes of apoptotic cell nuclei were observed by Hoechst staining. Cell growth and apoptosis of SH-SY5Y cells were determined by Western blotting and enzyme activity analysis of caspase-3, and AChE enzymatic activity was determined by the colorimetric assay. Results: In terms of cell viability and LDH release, exposure to FGE showed neuroprotective activities against 6-OHDA stimulated stress in SH-SY5Y cells. Furthermore, FGE reduced the 6-OHDA-induced apoptosis, as determined by Hoechst staining. The occurrence of apoptosis in 6-OHDA treated cells was confirmed by determining the caspase-3 activity. Exposure to 6-OHDA resulted in increased caspase-3 activity of SH-SY5Y cells, as compared to the unexposed group. However, pre-treatment with FGE inhibited the activity of caspase-3. The neuroprotective effects of FGE were also found to be caspase-dependent, based on reduction of caspase-3 activity. Exposure to FGE also inhibited the activity of AChE induced by 6-OHDA, in a dose-dependent manner. Conclusion: Taken together, our results show that FGE exhibits a neuroprotective effect in 6-OHDA treated SH-SY5Y cells, thereby making it a potential novel agent for the prevention or treatment of neurodegenerative disease.

Change in Nitrogen Compounds of Fermented Fodder for Sea Cucumber during Three Step Fermentation on Sludge (고형오물을 이용한 해삼용 3단 발효사료 제조 중 질소 성분 변화)

  • Lee, Su-Jeong;Ko, Yu-Jin;Kim, Eun-Ja;Kang, Seok-Jung;Ryu, Chung-Ho
    • Journal of agriculture & life science
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    • v.50 no.4
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    • pp.147-155
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    • 2016
  • This study presented a measure for turning by-products, released from land farming sites, into resources. The measure involved adding food by-products such as rice bran and nonfat soybean to the sludge, released from the eel farming sites, inoculating the lactic acid bacteria, Aspergillus oryzae, and Bacillus subtilis by step, fermenting them, and measuring the changed ingredients of the fermented fodder. The water content of the fermented fodder by the step of preparation was the first-step fermented product (14.6%) using the lactic acid bacteria, and the second and third-stage fermented product (33.0% and 34.0% respectively) using Aspergillus oryzae and Bacillus subtilis. The pH level was found to be 5.38 in the first-step fermented product due to the secretion of lactic acid caused by the lactic acid bacteria, and the pH level of the second and third-stage fermented products was 5.66 and 7.26, respectively, showing that the pH level increased. The phytic acid content was 0.126g/100g in the first-step fermented product, 0.004g/100g in the second-stage fermented product, and 0.093g/100g in the third-stage fermented product. The measurement of nitrogen content revealed that the amino nitrogen content was high with 1226.37mg% in the second-stage fermented product, and a little lower with 710.18mg% in the third-stage fermented product. The ammonium nitrogen content increased from 0.988mg/kg in the first-stage fermented product to 1.502mg/kg in the third-stage fermented product. Total nitrogen content increased to 2.78% in the first-stage fermented product, 4.08% in the second-stage fermented product, and 4.85% in the third-stage fermented product. As fermentation continued with the three microbes, the phytic acid decreased, and the protein decomposition rate increased. Also, due to the 3 step fermentation, the low-molecule nitrogen ingredient content increased, suggesting that the fodder was developed to offer high digestion and absorption.

Microbiological Studies of Korean Native Soy-sauce Fermentation -A Study on the Microflora Changes during Korean Native Soy-sauce Fermentation- (한국재래식(韓國在來式) 간장의 발효미생물(醱酵微生物)에 관(關)한 연구(硏究)(제2보(第二報)) -한국재래식(韓國在來式) 간장의 담금중(中)에 있어서의 발효미생물군(醱酵微生物群)의 소장(消長)에 관(關)한 연구(硏究)-)

  • Lee, Woo-Jin;Cho, Duck-Hiyon
    • Applied Biological Chemistry
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    • v.14 no.2
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    • pp.137-148
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    • 1971
  • Studies were carried out to investigate the main fermentation microorganisms and their flora changes during Korean native soy-sauce fermentation. Korean native Maeju loaves collected from 5 Do's were separated into surface and inner parts. Four different soy-sauces-the surface part Maeju fermented soy-sauce, the inner part, the surface and inner part combined Maeju fermented soy-sauce, and the semi-Japanese type soy-sauce were fermented and the changes of fermentation microorganism flora and the various chemical components during the period of their fermentations were studied. Besides, 14 home-made soy-sauces collected from 14 different places all over Korea were examined in comparison with the laboratory soy-sauces and to determine the characteristics of Korean native soy-sauce. The results were as follows: 1. The main microorganisms in Korean native soy-sauce fermentation were determined as; Aerobic bacteria: Bacillus subtilis, Bacillus pumilus Lactic acid bacteria: Pediococcus halophilus, Leuconostoc mesenteroides Yeasts: Torulopsis datila, Saccharomyces rouxii 2. Microflora changes during Korean native soy-sauce fermentation were as follows; Aerobic bacteria increased until the 2nd week of fermentation and then gradually decreased. The lactic acid bacteria increased until the 3rd week, after which decreased. When the lactic acid fermentation lowered the pH value to below the 5.4, yeasts were able to grow and participate the fermentation. As the production of organic acids amounted, to a certain height, the growth of all microorganisms lead to the period of decline or death at about the 2nd month of fermentation. After boiling of soy-sauce most microorganisms except a few of Bacillus sp. disappeared. Occosionally yeasts and lactic acid bacteria survived depending upon the composition of soy-sauce. 3. Changes of general chemical components influencing the microflora were investigated for the period of Korean native soy-sauce fermentation. Tetal acidity, salt concentration and total nitrogen were increasing steadily over the entire period of fermentation. pH values were dropping to a certain degree of about 4.5. Salt concentration and pH value seemed to be the important factors influencing the microflora. 4. The microflora were influenced by chemical components of soy-sauce. Aerobic bacteria were able to survive in all soy-sauce as they made spores. Growth of lactic acid bacteria was inhited at 23-26% of salt concentration and pH 4.8. Soy-sauce yeasts started to grow only at pH below 5.4 and seemed to be inhibited at around 26% of salt concentration under pH 4.5-4.7. 5. The open kettle boiling of soy-sauce, the characteristic process of Korean native soy-sauce manufacturing, was effective to sterilize microorganisms, increase the salt concentration, and coagulate proteins. 6. The average viable cell counts of microorganism found in collected samples of home-made Korean native soy-sauces were; Aerobic bacteria: $53{\times}10^2\;cell/ml$ Lactic acid bacteria: 34 cell/ml Yeasts: 14 cell/ml The average values of chemical compositions of samples of home-made Korean native soy-sauce were; Salt concentration: 28.9% pH value: 4.79 Total acidity(lactic acid): 0.91g/100ml Total nitrogen: 1.09g/100ml

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Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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A Study on the Manufacturing of Soysauce by the Combined use of Aspergillus sojae and Bacillus subtilis (단백질분해세균(蛋白質分解細菌)을 병용(倂用)한 간장제조(製造)에 관(關)한 연구(硏究))

  • Kim, Ze-Uook;Cho, Sung-Hwan
    • Applied Biological Chemistry
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    • v.18 no.1
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    • pp.1-9
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    • 1975
  • In order to study on the preparation of improved soysauce, Koji(soybean and wheat mash fermented by the strain Aspergillus sojae), Koji-Natto(mixed with the various proportion of Koji to Natto; 8 : 2, 6 : 4, 4 : 6, 2 : 8) and Natto (prepared using the strain: Bacillus subtilis) were prepared as soysauce materials. Sensory evaluation for the soysauces made from above-mentioned soysauce materials and the contents of total nitrogen, amino nitrogen, ammonia nitrogen, reducing sugar, total acid and enzyme activity were determined. The results were summarized as follows: 1. Protease activity of Natto was over twice higher than that of Koji and amylase activity of Koji was three times higher than that of Natto. Koji-Natto showed successive increase of amylase activity and successive decrease of protease activity as Koji proportion was increased to Natto. 2. The contents of total nitrogen and amino nitrogen in all the soysauce mashes displayed linear increases or no changes of these contents as aging. The contents of total nitrogen and amino nitrogen in the mahes were higher as Natto proportion increase to Koji during the aging period. 3. In Natto soysauce mash and Koji-Natto soysauce mashes the contents of ammonia nitrogen were gradually increased until $1{\sim}2$ months and rapidly reduced after that period, while Koji soysauce mashes were continuatively increased. 4. In all the soysauce mash the reducing sugar increased preeminently until the two months and after then there was a rapid decrease as aging the contents of reducing sugar in the mash were higher as Natto proportion increased to Koji during aging period. 5. The total acid of Koji and Koji-Natto (8 : 2) soysauce mash showed gradual increase as aging, while contents of total acid in Natto and other Koji-Natto soysauce mashes increased preeminently until 50${\sim}$70 days aging, after then decreased. 6. The results of sensory evaluation for three-month-fermented soysauces showed the most excellent in Koji-Natto (6 : 4) and successive order in the soysauces consisting of Koji and Natto proportion: 8 : 2, 10 : 0, 4 : 6, 2 : 8, 0 : 10.

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Antimicrobial Activity of Ethanol Extract from Sargassum thunbergii (지충이(Sargassum thunbergii) 에탄올 추출물의 항균활성)

  • Lee, So-Young;Song, Eu-Jin;Kim, Koth-Bong-Woo-Ri;Yoon, So-Young;Kim, Seo-Jin;Lee, So-Jeong;Hong, Yong-Ki;Lim, Sung-Mee;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.4
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    • pp.502-508
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    • 2009
  • Antimicrobial activity of Sargassum thunbergii was determined by paper disc assay and minimum concentration inhibitor (MIC) test. A water extract of S. thunbergii did not show the antimicrobial activity, but an ethanol extract of S. thunbergii (SHE) inhibited Serratia liquefaciens, Salmonella Typhimurium, Pseudomonas aerogenosa and all of the tested gram-positive bacteria at 4 mg/mL. Especially, Bacillus subtilis, Clostridium perfringens and Listeria monocytogenes were susceptible to SHE. As the results of MIC test, SHE inhibited the growth of B. subtilis, Staphylococcus aureus and Listeria monocytogenes at concentration of $0.1{\sim}0.3%$, and inhibited C. perfringens at 0.01%. In the thermal and pH stability test for SHE, antibacterial activities of SHE were maintained when the SHE was treated at $121^{\circ}C$ for 15 minutes or under pH $2{\sim}8$. SHE was partitioned in the order of n-hexane, chloroform, ethyl acetate and butanol. As the results of the MIC test for each obtained fraction, no fraction exhibited higher antibacterial activity than that of the crude SHE. However, a mixture of chloroform, ethylacetate and ethanol fractions showed higher antibacterial activity than SHE.

Antimicrobial Activity of the Sargassum fulvellum Ethanol Extract and the Effect of Temperature and pH on Their Activity (모자반 에탄올 추출물의 항균활성과 열 및 pH 안정성)

  • Yoon, So-Young;Lee, So-Young;Kim, Koth-Bong-Woo-Ri;Song, Eu-Jin;Lee, So-Jeong;Lee, Chung-Jo;Park, Na-Bi;Jung, Ji-Yeon;Kwak, Ji-Hee;Nam, Ki-Wan;Ahn, Dong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.42 no.2
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    • pp.155-159
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    • 2010
  • The antimicrobial activity of Sargassum fulvellum (SF) was investigated using the agar diffusion assay and MIC test. In addition, the stability of this activity under extreme heat and pH conditions was examined. The SF ethanol extract was shown to display strong antimicrobial activities against B. subtilis, C. perfringens, L. plantarum, S. aureus, L. monocytogenes, S. cerevisae and C. tropicalis in the agar diffusion assay at the concentration of 4 mg/mL. The MIC value of the SF ethanol extract against the tested microbes ranged from 0.05 to 0.0063%. In the heat and pH stability test, the antimicrobial activity of the SF ethanol extract was not altered when the temperature was maintained at $121^{\circ}C$ for 15 min, and it was also not affected in the pH range of 2-10. These results suggest that the SF ethanol extract is highly stable against drastic changes in temperature and pH.

Effect of antioxidation and antibacterial activity on crude extract and Characterization of American Cockroaches (Periplaneta americana L.) in Korea (국내 서식 미국바퀴(Periplaneta americana L.)의 특성 및 추출물의 항산화·항균 효과)

  • Kim, Jung-Eun;Kim, Seon-Gon;Kang, Sung-Ju;Kim, Chun-Sung;Choi, Yong-Soo
    • Journal of Sericultural and Entomological Science
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    • v.53 no.2
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    • pp.135-142
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    • 2015
  • The American cockroaches, Periplaneta americana L. was the most important worldwide pest species. It has been an public health problems. We were determinated life cycle and extraction of crude extracts by chemical reagents from cockraches (P. americana L.). The extracted crude solution has been antibacterial activity to gram negative bacteria (Pseudomonas aeruginosa, $6.44{\pm}1.03mm$), gram positive bacteria (Bacillus subtilis, $1.88{\pm}0.40mm$), and fungus (Candida albicans, $5.61{\pm}0.57mm$) using radial diffusion assay. We were analysed of up-regulation of Glutathione-S-transferases (GSTs) stimulation, indicating that antioxidantial protein from various classes are simultaneously expressed in a single insect upon infection or injury. The gene from Periplaneta americana L. were cloned, analysed sequence, and measured protein expression by Real Time PCR (Polymerase Chain Reaction).

Effect of Extracts and Bacteria from Korean Fermented Foods on the Control of Sesame Seed-Borne Fungal Diseases (발효식품 추출물과 미생물을 활용한 참깨 종자전염성 병 방제)

  • Kim, Yong-Ki;Hong, Sung-Jun;Shim, Chang-Ki;Kim, Min-Jeong;Park, Jong-Ho;Han, Eun-Jung;Park, Jong-Won;Park, So-Hyang;Jee, Hyeong-Jin;Kim, Seok-Cheol
    • Research in Plant Disease
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    • v.21 no.4
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    • pp.297-308
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    • 2015
  • In order to control seed-borne diseases, we obtained extracts from commercial fermented food products of Kimchi, Gochujang, Doenjang, Ganjang and Makgeolli and their suppressive effects against seed-borne diseases were studied. In addition, the suppressive effects of bacterial strains isolated from the fermented foods were screened in vitro and in vivo. Among fifty food extracts, twenty food-extracts suppressed more than 92% incidence of seedling rots in vitro and seven food extracts increased 58.3-66.8% of healthy seedling in the greenhouse. Among 218 isolates from the fermented foods, 29 isolates showing high antifungal activity against seven seed-borne fungal pathogens were selected. Among 29 isolates, 13 isolates significantly reduced seedling rot and increased healthy seedlings. Sixteen isolates with high antifungal activity and suppressive effect against sesame seedling rots were identified by 16S rRNA sequencing. Fourteen of sixteen isolates were identified as Bacillus spp. and the other two isolates from Makgeolli were identified as Saccharomyces cerevisiae. It was confirmed that B. amyloliquifaciens was majority in the effective bacterial population of Korean fermented foods. In addition, when the bioformulations of the two selected effective microorganisms, B. amyloliquifacien Gcj2-1 and B. amyloliquifacien Gcj3-1, were prepared in powder forms using bentonite, kaolin, talc and zeolite, talc- and kaolin-bioformulation showed high control efficacy against sesame seed-borne disease, followed by zeolite-bioformulation. Meanwhile control efficacy of each bentonite-bioformulation of B. amyloliquifacien Gcj2-1 and B. amyloliquifacien Gcj3-1 was lower than that of bacterial suspension of them. It was found that the selected effective microorganisms from Korean fermented foods were effective for controlling seed-borne diseases of sesame in vitro and in the greenhouse. We think that Korean fermented food extracts and useful microorganisms isolated from the extract can be used as bio-control agents for suppressing sesame seed-borne diseases based on above described results.