• 제목/요약/키워드: Shaking flask culture

검색결과 34건 처리시간 0.018초

Characterization of Endochitosanases-Producing Bacillus cereus P16

  • Jo, Yu-Young;Jo, Kyu-Jong;Jin, Yu-Lan;Jung, Woo-Jin;Kuk, Ju-Hee;Kim, Kil-Yong;Kim, Tae-Hwan;Park, Ro-Dong
    • Journal of Microbiology and Biotechnology
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    • 제13권6호
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    • pp.960-968
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    • 2003
  • A bacterial isolate showing a strong endochitosanase activity was isolated from soil and then characterized. The isolate was identified and designated as Bacillus cereus P16, based on morphological and biochemical properties, assimilation tests, cellular fatty acids pattern, along with 16S rRNA gene sequence. The optimized medium for producing extracellular chitosanase in a batch culture contained 1% tryptone, 0.5% chitosan, and 1% NaCl (pH 7.0). Powder chitosan and tryptone served the best as carbon and nitrogen sources, respectively, for the chitosanase production. Chitosanase activity was the highest when culture was completed at $37^{\circ}C$ among various temperatures ($20-42^{\circ}C$) tested in a shaking incubator (200 rpm). The levels of chitosanase activity in the culture fluid were 2.0 U/ml and 3.8 U/ml, respectively, when incubated in a flask for 60 h and in a jar fermenter for 24 h. The culture supernatant showed a strong liquefying activity on the soluble chitosan. The viscosity of 1% chitosan solution, that was incubated with the culture supernatant, was rapidly decreased, suggesting the secretion of endochitosanolytic enzymes by P16. The culture fluid revealed six endo-type chitosanase isozymes, two major (38 and 45 kD), and four minor (54, 65, 82, and 96 kD) forms by staining profile. The crude enzymes were very stable, and full activity was maintained for 4 weeks at $4^{\circ}C\;or\;-20^{\circ}C$ in the culture supernatant, suggesting a highly desirable stability rate for making an industrial application of the crude enzymes. The supernatant also cleaved the insoluble chitosan powder, but the hydrolysis rate was much lower. The enzymic degradation products of chitosan contained $(GlcN)_n$ (n=2-8). The concentration of chitosan in the reaction mixture of the crude enzyme affected the chitooligosaccharides composition of the hydrolysis products. When the higher concentration of chitosan was used, the higher degree of polymerized chitooligosaccharides were produced. By comparison with other commercial chitosanase preparations, P16 was indeed found to be a valuable enzyme source for industrial production of chitooligosaccharides from chitosan.

Micrococcus sp. M1에 의한 Phenol과 p-Creso의 생분해 Kinetics (Biodegradation Kinetics of Phenol and pcresol by Micrococcus sp. M1)

  • 손홍주;장웅석;이건;이상준
    • 한국환경과학회지
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    • 제6권2호
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    • pp.153-163
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    • 1997
  • In order to fad the most fitted biodegradation model, biodegradation kinetics model to the initial phenol and p-cresot concentrations were investigated and had been fitted by the linear regression. Bacteria capable of degrading p-cresol were isolated from soil by enrichment culture technique. Among them, strain Ml capable of degradillg p.rcresol has also degraded phenal and was identified as the genus Micrococcus from the results from of taxonomical studies. The optimal tonditlons for the biodegradation of phenal and p-cresol by Micrococcus sp. Ml were $NH_4NO_3$ 0.05%, pH 7.0, 3$0^{\circ}C$, respectively, and medium volume 100m1/250m1 shaking flask. iwicrococcus sp. Ml was able to grow on phenal concentration up to 14mM and p-cresol concelltration up to 0.8mM. With increasing substrate concentraction, the lag period increased, but the maximum specific growth rates decreased. The yield coefficient decreased with increasing substrate concentation. The biodegradation kinetics of phenol and p-cresol were best described by Monod with growth model for every experimented concentration. In cultivation of mixed substrate, p-cresol was degraded first and phenol was second. This result implies that p-cresol and phenol was not degraded simultaneously.

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초임계수 처리로 가수분해된 목질계 바이오매스를 이용한 바이오 에탄올 생산 (Bioethanol production from wood biomass hydrolysate with supercritical water treatment)

  • 서현범;한재건;최원석;이오규;이수민;최석환;이현용;정경환
    • KSBB Journal
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    • 제23권6호
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    • pp.494-498
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    • 2008
  • 초임계수 처리를 통하여 얻어진 목질계 바이오매스 가수분해 물을 이용한 바이오 에탄올 생산에 대하여 연구하였다. 초임계수 처리 가수분해물은 바이오에탄올 생산을 위한 배지의 탄소원으로 사용되었다. 농축된 초임계수 처리 가수분해물 (SCW3)을 사용하여 효모를 배양하였을 때, 다른 두 가지 초임계수 처리 가수분해물 (SCW1, SCW2) 을 사용한 경우에 비하여 효모의 성장속도가 늦었다. 그리고 모든 경우에 0.51에서 0.56 (%, w/v)의 바이오 에탄올이 생산되었다. 그래서 농축된 초임계수 처리 가수분해물 (SCW3)을 활성탄과 수산화 칼슘으로 전처리하여 페놀류 독성물질을 제거하였다. 활성탄 전처리가 보다 효과적으로 94.6%의 페놀류 화합물을 제거하였고, 바이오 에탄올도 0.96 (%, w/v) 생산 할 수 있게 하였고, 환원당을 기준으로 한 바이오에탄올 수율도 0.5에 이르렀다.

카탈라제를 생산하는 고초균 (Bacillus sp.)의 고정화 및 과산화수소 분해 특성 (Immobilization of Bacillus sp. Strains, Catalase Producing Bacteria and Their Hydrogen Peroxide Removal Characteristics)

  • 한경아;장윤희;이종일
    • KSBB Journal
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    • 제25권6호
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    • pp.520-526
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    • 2010
  • 토양 미생물로부터 스크리닝한 미생물 중 카탈라제 생산량이 높은 세 가지 미생물 (Bacillaceae bacterium BKBChE-1, Bacillus sp. BKBChE-2와 Bacillus flexus BKBChE-3)을 배양하여 얻은 카탈라제 효소가 과산화수소를 분해하는 것을 확인하고, 미생물 균체를 천연제올라이트에 정전기적 흡착법으로 고정하였다. 주사전자 현미경을 통해 각각의 미생물이 지지체에 잘 흡착된 것이 관찰되었고, 10.5 mM의 과산화수소를 1시간 이내에 완전히 분해되는 것을 확인하였다. 과산화수소 분해 속도가 가장 빠른 Bacillus flexus BKBChE-3은 온도와 pH 영향성 실험을 통해 고온과 pH 10 이상의 강알칼리 조건에서도 30일 이후까지 미생물이 생존하는 것을 관찰하였다. 또한 고정화된 미생물의 경우 온도 $25-40^{\circ}C$, pH 7-10의 조건에서 생균수가 계속하여 증가하는 것을 확인하였다. 본 실험을 통해 Bacillus flexus BKBChE-3이 생산한 카탈라제 효소 뿐만 아니라 고정화된 균체도 과산화수소의 제거에 효과적임을 확인할 수 있었다. 이들 결과로부터 미생물 발효를 통해 생산된 효소의 생산량 증가와 미생물 개체 수 증가를 이용하여 산업폐수에 적용하면, 과산화수소의 제거 효율을 높이고 균체 재활용을 통한 공정비용을 절감할 수 있을 것으로 사료된다.