• Title/Summary/Keyword: Bacillus sp. K91

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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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Development of Optimal Culture Media for the Stable Production of Mushroom (버섯의 안정적 생산을 위한 최적배지의 개발)

  • Gal, Sang-Wan;Lee, Sang-Won
    • Applied Biological Chemistry
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    • v.45 no.2
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    • pp.71-76
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    • 2002
  • Several antagonistic bacteria, SD-1, 4, 10, 11, 14, 15, and 16, which have strong CMCase and amylase activities, were isolated from the fermented mushroom media. Among them, SD-1, 10, 11, and 15 have strong antibacterial activities against the mushroom pathogenic bacteria Pseudomonas sp., and SD-1, 10, 11, 14, and 16 have strong antifungal activities against the mushroom pathogenic fungi, Trichoderma sp. SD-14, 15, and 16 did not inhibit the growth of mushroom Pleurotus eryngii ASI-2302, and Pleurotus ostreatus ASI-2042 and ASI-2180. When the culture broth mixture of the seven bacterial strains was applied to the mushroom media, the growths of pathogens, Pseudomonas sp. and Trichoderma sp., were inhibited.

Antimicrobial activities of oral bacteria by lichen extracts (지의류 추출물의 구강세균에 대한 항균효과)

  • Kim, Eun-Mi;Cho, Min-Jeong
    • Journal of Korean society of Dental Hygiene
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    • v.12 no.1
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    • pp.81-91
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    • 2012
  • Objectives : By investigating antimicrobial activity of natural extracts, identifying its usefulness as antibiotic material to oral bacteria. Methods : Antimicrobial activity tests of 25 natural extracts were implemented on 6 type strains alleged to cause dental caries and 10 clinical strains isolated and identified from dental caries. Results : Among medicinal plants, Coptis japonica, Scutellaria baicalensis, Dictamnus dasycarpus showed antimicrobial activity. Among lichens, 3 methanol extracts and 6 acetone extracts showed antimicrobial activity. Minimum Inhibitory Concentration(MIC) test of Usnea aurantiacoatra KoLRI 004184, an acetone extract of lichen with the highest antimicrobial activity, on 6 type strains involved in dental caries and 10 strains isolated from dental caries resulted as follows; Actinomyces oris 1041 $12{\mu}g/ml$, for Corynebacterium durum 3151 $13{\mu}g/ml$, for Rothia dentocariosa KCTC $3204^T$ $14{\mu}g/ml$, for R. dentocariosa 911 $15{\mu}g/ml$, for R. dentocariosa 1822 $12{\mu}g/ml$, for Lacto-bacillus casei KACC $12413^T$ $12{\mu}g/ml$, for L. acidophilus KACC $12419^T$ $16{\mu}g/ml$, for L. rhamnosus 2421 $15{\mu}g/ml$, for Streptococcus mutans KCTC $3065^T$ $15{\mu}g/ml$, for S. mutans 121 $13{\mu}g/ml$, for Streptococcus oralis 2221 $18{\mu}g/ml$, for S. salivarius KCTC 5512 $14{\mu}g/ml$, for S. salivarius 122 $12{\mu}g/ml$, for S. sanguinis KCTC $3284^T$ $14{\mu}g/ml$, for S. sanguinis 912 $14{\mu}g/ml$, for Neisseria sp. KEM232 $12{\mu}g/ml$. Conclusions : Even a small amount of extract from lichen Usnea aurantiacoatra KoLRI 004184 showed very high level of antimicrobial activation against all 16 bacterial strains involved in dental caries.

Synergistic Action Modes of Arabinan Degradation by Exo- and Endo-Arabinosyl Hydrolases

  • Park, Jung-Mi;Jang, Myoung-Uoon;Oh, Gyo Won;Lee, Eun-Hee;Kang, Jung-Hyun;Song, Yeong-Bok;Han, Nam Soo;Kim, Tae-Jip
    • Journal of Microbiology and Biotechnology
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    • v.25 no.2
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    • pp.227-233
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    • 2015
  • Two recombinant arabinosyl hydrolases, α-L-arabinofuranosidase from Geobacillus sp. KCTC 3012 (GAFase) and endo-(1,5)-α-L-arabinanase from Bacillus licheniformis DSM13 (BlABNase), were overexpressed in Escherichia coli, and their synergistic modes of action against sugar beet (branched) arabinan were investigated. Whereas GAFase hydrolyzed 35.9% of L-arabinose residues from sugar beet (branched) arabinan, endo-action of BlABNase released only 0.5% of L-arabinose owing to its extremely low accessibility towards branched arabinan. Interestingly, the simultaneous treatment of GAFase and BlABNase could liberate approximately 91.2% of L-arabinose from arabinan, which was significantly higher than any single exo-enzyme treatment (35.9%) or even stepwise exo- after endo-enzyme treatment (75.5%). Based on their unique modes of action, both exo- and endo-arabinosyl hydrolases can work in concert to catalyze the hydrolysis of arabinan to L-arabinose. At the early stage in arabinan degradation, exo-acting GAFase could remove the terminal arabinose branches to generate debranched arabinan, which could be successively hydrolyzed into arabinooligosaccharides via the endo-action of BlABNase. At the final stage, the simultaneous actions of exo- and endo-hydrolases could synergistically accelerate the L-arabinose production with high conversion yield.

Effect of Biozyme on Alcohol Metabolism

  • Park, Seung-Hee;Nam, Suk-Woo;Sungpil Yoon;Minsik Son;Nam, Tae-Kyun;Seo, Dong-Wan;Sungyoul Hong;Lee, Hyang-Woo
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.91-91
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    • 1995
  • 에탄올에 의한 급성독성은 에탄올, 아세트알데히드 및 에탄을 대사산물의 변형생성물질 등에 기인하므로 알콜 섭취 후 혈중 에탄을 농도 및 아세트알데히드의 농도를 낮추는 것은 음주에 의한 급성 중독상태에 머무는 시간을 단축시키는데 중요하다. 본 실험에서는 Bacillus subtilis natto sp.를 식물 추출액을 배지로 하여 배양한 후 단백 분해효소로 고분자물질을 절단하여 얻은 발효액인 바이오짐(상품명: Biozyme)을 주성분으로 한 '비지니스(조선무약합자회사)' 의 인체 혈중 알콜대사 촉진작용을 검색하였으며, 비지니스 및 바이오짐이 랫드의 에탄을 대사에 미치는 효과를 검토하여 다음과 같은 결과를 얻었다. 인체 혈중 알콜대사 촉진작용을 검색하기 위해 알콜 섭취 전과 후의 혈중 에탄올 농도를 비교하였는데, 대조군에 비해 비지니스 투여군이 30분 후부터 2시간 후까지 혈청 알콜농도가 낮게 나타났다. 대조군의 혈청 알콜농도를 100으로 하였을 때 알콜 투여 30분, 60분, 90분, 120분 경과 후 시험군은 각각 대조군의 84.3%, 89,0%, 85.9%, 75.8%를 나타내어 평균 16% 정도 낮았다. 시험군의 AUC는 대조군의 AUC의 89%로 비지니스 투여군에서 혈액내 알콜의 제거가 빠르게 진전된다는 것을 보여주었다. 또한, 랫드의 에탄을 대사에 미치는 효과를 알아보고자 바이오짐 및 비지니스 투여 후 채혈하여 혈중 에탄을 및 아세트알데히드 농도를 측정하였다. 비즈니스 투여시 혈중 알콜 농도는 알콜 투여 60분 경과후 가장 큰 감소 효과(대조군:83.70$\pm$11.80mg/이, 시험군:45.12$\pm$6.63mg/d1, 47% 감소)를 나타내었으며, 시험군의 AUC는 대조군에 비해 30% 감소하였다. 혈중 아세트알데히드 농도는 투여 60분 후 비지니스 투여군(4.56$\pm$0.51nmol/$m\ell$)이 대조군(6.45$\pm$0.64nmo1/$m\ell$)에 비해 유의성 있는 감소(29%)를 나타내었으며, 시험군의 AUC는 대조군에 비해 35% 감소하였다. 바이오짐 투여시 혈중 에탄을 농도가 알콜 투여 2시간 경과 후 가장 큰 감소 효과(대조군:49.10$\pm$5.20mg/dl, 시험군:25.90$\pm$7.16mg/d1, 47% 감소)를 나타내었으며, 시험군의 AUC는 대조군에 비해 39% 감소하였고, 혈중 아세트알데히드의 농도는 투여 60분후 시험군(3.96$\pm$0.07nmo1/$m\ell$)이 대조군(6.45$\pm$0,64nmo1/$m\ell$)에 비해 유의성 있는 감소(39%)를 나타내었으며, 시험군의 AUC는 대조군에 비해 48% 감소하였다 한편, 시험관내 에탄올 대사 효소에 대한 바이오짐의 효과를 검색해본 결과 바이오짐(2.0 $\mu\textrm{g}$/assay)에 의해 Aldehyde dehydrogenase(1.5unit/assay)의 활성이 14% 증가되었다. 본 연구의 결과로 볼 때, 비지니스 및 바이오짐은 음주 후 상승된 혈중 에탄을 농도 및 아세트알데히드의 농도를 현저히 감소시키는 효과가 있었다.

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Hypoglycemic and Antioxidative Effects of Fermented Chaga Mushroom (Inonotus obliquus) on Streptozotocin-induced Diabetic Rats (Streptozotocin 유발 당뇨쥐에서 발효 차가버섯의 항당뇨 및 항산화 효과)

  • Cha Jae-Young;Jun Bang-Sil;Lee Chi-Hyeoung;Yooi Ki-Soo;Moon Jae-Chul;Cho Young-Su
    • Journal of Life Science
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    • v.15 no.5 s.72
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    • pp.809-818
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    • 2005
  • The effects of fermented chaga mushroom (Inonotus obliquus) on the concentrations of serum glucose, insulin, lipids and lipid peroxidation in streptozotocin (STZ)-induced diabetic rats were investigated. Rats were fed a semisynthetic diet supplemented with 50 g/kg chaga mushroom powder (the CM group) and fermented chaga mushroom powder (the FCM group), and no supplemented (the control group) for 3 weeks. The polysaccharide concentrations were CM by $42.9\%$ and FCM by $ 39.1\%$, and the total polyphenol concentrations were CM by $ 0.80\%$ and FCM by $0.91\%$. Feed intakes and water consumption, serum glucose, insulin, triglyceride, and blood urea nitrogen concentrations were significantly lower in the FCM group than in both the CM and control groups. The activities of AST and ALT were also significantly lower in the FCM group than in the control group. No significant differences were detected with regard to the serum cholesterol and creatinine concentrations among the experimental groups. Lipid peroxidations in hepatic homogenate, microsomal and mitochondrial subcellular and pancreas were significantly lowered by the administration of FCM in the STZ-diabetic rats. Hepatic glutathione concentrations, which is closely associated with antioxidant system, was significantly higher in the FCM group than in the control group, indicating a marked effect of FCM administration on the endogenous antioxidant system. However, CM treatment showed a moderate antioxidative activity in the STZ-diabetic rats. Our results indicate that fermented chaga mushroom exert hypoglycemic and antioxidative effects in type 1 diabetes mellitus.

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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