• 제목/요약/키워드: Bacillus sp. 79-23

검색결과 6건 처리시간 0.017초

Cellulase를 생산하는 Bacillus sp. 79-23 분리와 효소 생산성

  • 윤기홍;정경화;박승환
    • 한국미생물·생명공학회지
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    • 제25권6호
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    • pp.546-551
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    • 1997
  • A bacterium producing the extracellular cellulases was isolated from soil and has been identified as Bacillus sp. The isolate, named Bacillus sp. 79-23, was shown to be very similar to B. subtilis on the basis of its biochemical properties. The carboxymethyl cellulase (CMCase) of culture supernatant was most active at 60$\circ$C and pH 6.0, and retained 90% of its maximum activity at pH 7.0. The additional carbon sources affected the CMCase productivity than nitrogen sources in the culture medium. The carbon sources including wheat bran, rice straw, maltose and glucose increased the enzyme productivty. Especially, the maximum CMCase production was 5.2 units/ml in LB medium supplemented with 3% (w/v) wheat bran, which was 13-folds more than that in LB medium. It was found that the enzyme production was in association with the growth of Bacillius sp. 79-23. But, whean bran did not affect the growth of isolate, suggesting that increasement of CMCase production was owing to the induction of CMCase biosynthesis by wheat bran. In addition, both water-soluble and insoluble components of wheat bran was involved in induction of CMCase biosynthesis.

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Hyper-CMCase-Producing Mutants of Bacillus sp. 79-23 Induced by Gamma- Radiation

  • Yoon, Ki-Hong;Shin, In-Kyung;Jung, Kyung-Hwa;Park, Seung-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제9권4호
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    • pp.518-521
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    • 1999
  • Bacillus sp. 79-23 spores were irradiated with $^{60}Co$ gamma-rays at doses ranging from 0.5 to 5 kGy. Following gamma-irradiation, seven mutant strains were isolated by scoring the halo sizes formed around the colonies grown on LB agar plates containing 4% carboxymethylcellulose (CMC) and trypan blue. The mutant strains showed a 1.5 to 2-fold increase in carboxymethylcellulase (CMCase) activity over the parent strain. Wheat bran acted as an effective inducer for CMCase production in the parent and mutant strains. Mutant strains 68 and 70 were identified as exhibiting higher CMCase activities than those of other mutants in LB media both with and without 3% wheat bran. In addition, these strains seem to produce substantially lower amounts of capsular materials, whereas the parent strain produced large amounts of them in both liquid and solid LB media. In flask cultures, the CMCase production by mutants 68 and 70 reached maximum levels of 17.5 unit/ml and 15.7 unit/ml, respectively, in an LB medium containing 3% wheat bran.

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한국산 꽃소금과 천일염의 이화학적 특성 및 미생물 분석 (Physicochemical Properties and Microbial Analysis of Korean Solar Salt and Flower of Salt)

  • 이혜미;이우경;진중현;김인철
    • 한국식품영양과학회지
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    • 제42권7호
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    • pp.1115-1124
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    • 2013
  • 한국산 꽃소금의 특성을 알아보기 위하여 국내에서 생산된 꽃소금 3종과 천일염 1종의 이화학적 및 미생물 분석을 실시하였다. 수분함량은 $10.54{\pm}0.10{\sim}13.82{\pm}0.12%$, 나머지 조단백, 조지방, 조섬유는 거의 존재하지 않았다. NaCl은 신의도산 꽃소금이 $78.81{\pm}0.28%$, 비금산 꽃소금이 $81.67{\pm}0.34%$, 도초산 꽃소금 $84.61{\pm}0.21%$, 도초산 천일염이 $80.82{\pm}0.17%$로 나타났다. 불용분, 사분은 각각 0.01~0.05%, 0.01~0.03%로 낮게 검출되었다. 미네랄 분석은 도초산 꽃소금에서 K과 Mg 함량이 2,975.23 mg/kg, 9,886.72 mg/kg으로 비교적 낮게 나타났다. Ca은 도초산 소금 2종이 945.53 mg/kg, 942.43 mg/kg으로 낮은 함량을 보였다. 중금속 As, Cd, Pb, Hg은 모두 규격 이하로 검출되었다. 소금결정을 관찰한 결과로 천일염과 꽃소금 모두 핵이 중복되어 겹으로 적층된 것을 확인하였으며 크기는 비금산 꽃소금이 $0.067{\times}0.067mm$로 가장 작고 도초산 천일염이 $0.112{\times}0.124mm$로 꽃소금에 비해 크게 나타났다. 소금의 색도는 꽃소금의 L값이 천일염에 비해 높아 밝게 확인되었다. 관능검사 결과로 짠맛은 NaCl 함량과 유사하며, 쓴맛은 K과 Mg 함량이 적은 도초산 꽃소금이 낮게 나타났다. 천일염에 비해 꽃소금의 단맛과 기호도가 더 높은 것으로 확인되었다. 호염균 동정 결과 19종 모두 Firmicutes로 꽃소금에서 Marinibacillus, Paenibacillus, Bacillus 속이 12종으로 확인되며, 천일염은 Planococcus, Staphylococcus, Bacillus 속 3종으로 검출되었다. DGGE 실험 결과로 도초산 꽃소금에서 16개의 band와 도초산 천일염에서 15개의 band를 확인하였다. 동정 결과 도초산 꽃소금에서 Cupriavidus sp. ATHA3(14.43%), Cupriavidus sp. TSA5(10.41%), Maritimibacter sp. YCSD61-2(8.13%), uncultured bacteria(68%)로 확인되었고, 도초산 천일염에서는 Dunaliella salina(4.76%), Cupriavidus sp. ATHA3(15.80%), uncultured Mycoplasmataceae bacteria(51.72%), uncultured bacteria(27%)로 확인되었다.

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)- (Microbiological Studies of Korean Native Soy-sauce Fermentation -A Study on the Microflora Changes during Korean Native Soy-sauce Fermentation-)

  • 이우진;조덕현
    • Applied Biological Chemistry
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    • 제14권2호
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    • pp.137-148
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    • 1971
  • 한국재래식(韓國在來式) 메주를 전국(全國) 5개도(個道)에서 수집(收集)하여 메주 표면부분(表面部分), 내부부분(內部部分), 재래식(在來式) 메주 및 일본식(日本式) 방법(方法)을 도입(導入)한 개량식(改良式) 메주로 간장을 담그어 담금 중의 미생물(微生物) 소장(消長)에 관(關)하여 연구(硏究)하고 이들 미생물(微生物)의 flora에 영향(影響)을 주는 pH, 총산(總酸), 식염농도(食鹽濃度), 총질소(總窒素)를 조사(調査)하였다. 아울러 한국전성(韓國全城) 14개(個) 지방(地方)에서 현재(現在) 식용중(食用中)인 재래식(在來式) 간장을 수집(收集)하여 microflora 및 화학적성분(化學的成分)을 측정(測定)하여 다음과 같은 결과(結果)를 얻었다. 1. 한국재래식(韓國在來式) 간장의 주발효미생물군(主醱酵微生物群)을 구명(究明)한 결과 호기성(好氣性) 세균군(細菌群)은 Bacillus subtilis와 Bacillus pumilus 유산균군(乳酸菌群)은 Pediococcus halophilus 및 Leuconostoc mesenteroides, 그리고 효모군(酵母群)은 Rhodotorula flava, Torulopsis datila 및 Saccaromyces rouxii이었다. 2. 한국재래식(韓國在來式) 간장, 담금 중의 발효미생물(醱酵微生物)의 소장(消長)은 먼저 호기성(好氣性) 세균(細菌)이 담근후 2주(週)까지 증가(增加)하고 다음 호기성(好氣性)이 3주(週)쯤에 최고(最高)의 증식(增殖)을 보여 유균(乳菌) 등(等)의 유기산함량(有機酸含量)을 높이고 pH를 5,4부근(附近)으로 강하(降下)시킨 다음에 효모(酵母)가 증식(增殖)하기 시작(始作)하여 발효(醱酵)에 참여하고 있다. 유산균(乳酸菌)은 생산(生酸)에 의(依)해 자신(自身)과 호기성(好氣性) 細菌)과 생육(生育)을 조해(阻害)하고 있으며 담금 2개월(個月) 후(後)엔 모든 발효미생물(醱酵微生物)이 감퇴기(減退期) 내지(乃至) 사감기(死減期)에 접어들고 달인 뒤에는 대부분(大部分) 사감(死減)하고 소수(少數) Bacillus sp.만이 잔존(殘存)하나 담금액(液)의 조성(組成)에 따라 효모(酵母)와 유산균(乳酸菌)이 극소수(極少數) 잔존(殘存)하고 있는 경우도 있었다. 3. 담금중의 microflora에 영향(影響)을 주는 성분(成分)의 변화(變化)를 조사(調査)한바 간장의 전(全)담금 기간(其間)을 통(通)하여 총산(總酸), 식염농도(食鹽濃度) 및 총질소(總窒素)는 점차 증가(增加)하고 있으며 pH는 약(約) 4.5 정도(程度)까지 강하(降下)하고 있다. 4. Microflora는 주(主)로 식염농도(食鹽濃度)와 pH에 지배(支配)되는 것 같았다. 호기성세균(好氣性細菌)은 포자(胞子)의 형성(形成)으로 가장 생존력(生存力)이 강(强)한 것 같으며 유산균(乳酸菌)은 식염농도(食鹽濃度) $23{\sim}26%$, pH 4.8에서 생육(生育)이 조해(阻害)되었으며 효모(酵母)는 담금액(液)의 pH가 5.4 이하(以下)에서 비로소 발육(發育)을 시작(始作)하였고 pH $4.5{\sim}4.7$에서의 한계식염농도(限界食鹽濃度)는 26% 전후(前後)로 보였다. 5. 한국재래식(韓國在來式) 간장 재활과정중(製活過程中)의 지징(持徵)인 장시간(長時間)의 장달이기는 본(本) 실험결과(實驗結果)에서 보면 살균(殺菌) 단백질(蛋白質)의 응고(凝固) 식염(食鹽) 및 기지(基地) 성분(成分)의 농축(濃縮) 등(等)에 있어서 매우 효과적(效果的)인 방법(方法)이라는 것이 명백(明白)하다. 달인 후(後)의 간장의 숙성(熟成)은 미생물학적(微生物學的)이라기 보다는 오히려 화학적(化學的) 숙성(熟成)이 주(主)가 되는 것이라고 인정(認定)되었다. 6. 전국(全國)에서 수집(收集)한 한국재래식(韓國在來式) 간장의 발효미생물(醱酵微生物)의 평균생균수(平均生菌數)는 호기성세균(好氣性細菌) $53{\times}10^2$개/ml, 유산균(乳酸菌) 34개/ml, 효모(酵母) 14개/ml이었고 제화학성분(諸化學成分)의 평균치(平均値)는 식염농도(食鹽濃度) 28.9%, pH=4.79, 총산(總酸)(유산(乳酸)) 0.91g/100ml, 총질산(總窒酸) 1.09g/100ml이었다.

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Bacillus subtilis HJ18-3과 KACC 15935를 이용하여 제조한 청국장의 품질특성과 isoflavone 함량의 변화 (Changes in isoflavone content and quality characteristics of Cheonggukjang prepared with Bacillus subtilis HJ18-3 and KACC 15935)

  • 이경하;최혜선;최윤희;박신영;송진
    • 한국식품저장유통학회지
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    • 제21권1호
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    • pp.121-128
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    • 2014
  • 청국장 제조 시 사용되는 균주인 Bacillus sp. HJ18-3과 B. subtilis KACC 15935균주를 청국장 제조 시 starter로 접종하여 발효시켜 품질특성과 isoflavone 비배당체 함량을 측정하였다. a-amylase와 cellulase활성은 control과 B. subtilis HJ18-3 균주를 접종한 청국장이 B. subtilis KACC 15935 균주를 접종한 청국장보다 각각 2.56, 2.59배 그리고 1.13, 1.23배 높은 결과를 보였다. 그러나 protease활성은 B. subtilis KACC 15935과 B. subtilis HJ18-3의 활성이 높은 효소활성을 보였다. 아미노태질소 함량은 Control, B. subtilis KACC 15935, B. subtilis HJ18-3의 함량이 각각 $88.20{\pm}13.86$, $129.62{\pm}1.15$, $118.40{\pm}48.65mg%$로 나타났으며 이는 protease 효소활성의 결과와 유사한 경향을 보였다. 환원당 함량은 control, KACC 15935, HJ18-3의 함량이 각각 $1.95{\pm}0.72$, $2.62{\pm}1.17$, $2.59{\pm}0.76%$로 전반적으로 control에 비해 처리구가 더 높은 경향을 나타냈다. 호기적 총균수와 미비한 차이로 B. subtilis HJ18-3의 균수가 적은 경향을 나타낸 것은B. cereus와Candida albicans에 대해 항균력이 우수하다고 보고된 영향으로 해석된다. control과B. subtilis KACC15935, B. subtilis HJ18-3의 isoflavone 비배당체 형태인 daidzein과 genistein, glycitein을 합한 함량은 각각 $158.20{\pm}6.55$, $131.60{\pm}4.00$, $200.79{\pm}7.54ug/g$으로 나타났다. 이는 초기 raw soybean의 aglycone함량이 3.96 ug/g인 것에 비하여 39~50배 전 환율을 보인다. 또한 ${\beta}$-glucosidase활성이 있는 B. subtilis HJ18-3균을 접종하여 제조한 청국장이 control과 Kacc15935균을 접종하여 제조한 청국장보다 1.27, 1.53배 더 높은 aglycone 전 환율을 보였다. 따라서 본 연구에서 사용한 메밀 속성 장으로부터 분리된 B. subtilis HJ18-3 균주는 amylase, protease, cellulase와 같은 세포의 효소분비능이 우수하며, 아미노태질소와 환원당 함량이 높고, isoflavone 비배당체 함량을 증가시키는 기능을 함유한 균주로 확인되었으며, 앞으로도 우리나라 전통 청국장의 세계화를 위해 유용 starter로 찾는 노력이 더 필요할 것이다.

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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    • 한국균학회 2015년도 춘계학술대회 및 임시총회
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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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