• 제목/요약/키워드: cyclodextrin glucanotransferase

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Cyclodextrin Glucanotransferase의 열안정성 증가 (Increase of the Thermostability of Cyclodextrin Glucanotransferase)

  • 김진현;홍승서;이현수
    • KSBB Journal
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    • 제16권2호
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    • pp.212-215
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    • 2001
  • The effect of various additives on the thermostability of Bacillus sp. cyclodextrin glucanotransferase (CGTase) was investigated. CaCl$_2$, starch, and glycerol had a positive effect on the thermostability of the CGTase, which was very stable for 6 months with added starch (5%, w/v) and CaCl$_2$(0.05 M) at 30$^{\circ}C$.

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Bacillus stearothermophilus KJ16이 생산하는 Cyclodextrin Glucanotransferase 의 정제와 효소특성 (Purification and Characterization of Cyclodextrin Glucanotransferase from Bacillus stearothermophilus KJ16)

  • 권현주;남수완;김광현;송승구;윤종원;김병우
    • 생명과학회지
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    • 제8권3호
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    • pp.326-332
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    • 1998
  • Cyclodextrin glucanotransferase from B. stearothermophilus KJ16 that can produce both cyclodextrin glucanotransferase and cyclodextrinase was purified by ammonium sulfate precipitation, DEAE-cellulose chromatography, Sephadex G-100 chromatography, and FPLC. The molecular weight of the purifice enzyme was about 65,000 dalton by SDS-PAGE. The optimal pH and temperature were 6.0 and $60^{\circ}C$, respectively. The enzyme was stable at $50^{\circ}C$ for 1 hr and in the pH range of 5.5 and 8.5. Mercaptoethanol and dithiothreitol inhibited the enzyme activity strongly. The enzyme produced 60% cyclodextrin(CD) from 5% soluble starch with the $^{\alpha}$, $^{\beta}$, $^{\gamma}$-CD ratio of 42:46:12. Amylopectin was the most suitable substrate with 67% conversion to CD.

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Affinity Chromatography를 이용한 Cyclodextrin Glucanotransferase의 정제 (Purification of Cyclodextrin Glucanotransferase by Affinity Chromatography)

  • 안중훈;황진봉;김승호;김경은
    • 한국미생물·생명공학회지
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    • 제19권3호
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    • pp.313-314
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    • 1991
  • Bacillus stearothermophilus의 mutant가 생산하는 cyclodextrin glucanotransferase(CGTase)를 affinity chromatography법을 이용하여 정제하였다. 이 CGTase의 회수율은 95%이었고 specific activity는 26.2U/mg protein에서 485.5U/mg protein으로 증가하였다. 정제된 CGTase는 SDS-polyacrylamide gel 전기영동 결과 단일 band로 나타났다. CGTase는 affinity chromatography를 이용하여 one-step으로 정제할 수 있었다.

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여러 첨가물의 용매가 Bacillus stearothermophilus가 생산하는 Cyclodextrin Glucanotransferase의 열안정성에 미치는 영향 (Effect of Various Additives and Solvents on Thermostability of Cyclodextrin Glucanotransferase from Bacillus stearothermophilus)

  • 안중훈;황진봉;김승호
    • 한국미생물·생명공학회지
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    • 제19권4호
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    • pp.368-371
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    • 1991
  • Ethylene glycol, glycerol, sorbitol 그리고 sucrose가 Bacillus stearothermophilus가 생산하는 cyclodextrin glucanotransferase(CGYase)의 열안정성에 미치는 영향을 조사하였다. Glycerol, sorbitol 그리고 sucrose가 CGTase의 열안정성에 효과가 있었다. 이 효과는 첨가물의 농도와 밀접한 관계가 있었다. n-Butanol, 1,4-dioxane그리고 n-octane과 같은 유기 용매에서 CGTase의 열안정성을 조사하였다. 1,4-dioxane 과 n-octane은 CGTase의 열안정성을 증가시켰다. 특히, n-octane에서 CGTase를 $75^{\circ}C$에서 90분간 정치시킨 후에도 원래 효소활성의 81를 유지하였다.

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Bacillus stearothermophilus가 생산하는 Cyclodextrin Glucanotransferase: Affinity Chromatography를 이용한 정제 및 성질 (Cyclodextrin Glucanotransferase from Bacillus stearothermophilus:Purification by Affinity Chromatography and Its Properties)

  • 안중훈;황진봉;김승호
    • 한국미생물·생명공학회지
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    • 제18권6호
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    • pp.585-590
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    • 1990
  • Bacillus stearothermophilus가 생산하는 cyclodextrin glucanotransferase (CGTase)를 ammonium sulfate 법과 affinity chromatography법에 의해 정제하였다. 이 CGTase의 비활성은 11.5U/mg protein에서 33445.0U/mg protein으로 증가하였다. 이 효소의 분자량을 측정하기 위해 SDS-PAGE를 실시한 결과 분자량은 약 7,800이었다. 이 효소의 최적 pH와 온도는 각각 6.0과 $60^{\circ}C$이었다. 이 효소는 pH5.5와 10.0에서 안정하였다. 이 효소에 calcium ion을 첨가하였더니 열안정성이 증가하였다. 이 효소의 등전점은 약 4.8 이었다.

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분쇄마찰매체 불균일상 효소반응계를 활용한 생전분을 당공여체로 하는 Cyclodextrin Glucanotransferase의 당전이 반응 (Transglycosylation Reaction of Cyclodextrin Glucanotransferase in the Attrition Coupled Reaction System using Raw Starch as a Donor)

  • 이용현;백승걸;박동찬;신현동
    • 한국미생물·생명공학회지
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    • 제21권5호
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    • pp.461-467
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    • 1993
  • Transglycosylation reaction of cyclodextrin glucanotransferase (CGTase) was analyzed in the attrition coupled heterogeneous reaction system using raw starch as a donor` and mono-, di-saccharide, and glycoside as acceptors. For transglycosylation reaction of stevioside, the transglycosylation rate was similar and the transglycosylation yield was increased compare with conventional process using liquefied starch as the donor. Also the accumulation of maltooligosaccharides in reaction mixture was minimized.

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Saccharomyces cerevisiae에 발현된 재조합 cyclodextrin glucanotransferase의 생화학적 특성 (Biochemical Properties of Recombinant Cyclodextrin Glucanotransferase Expressed in Saccharomyces cerevisiae)

  • 박현이;남수완;김병우
    • 생명과학회지
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    • 제11권3호
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    • pp.230-234
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    • 2001
  • The cyclodextrin glucanotransferase(CGTase) gene of Bacillus macerans was expressed in Saccharomyces cerevisiae and the recombinant CGTase was partially purified from the yeast culture supernatant. The optimal pH and temperature of the CGTase were found to be 6.0 and 5$0^{\circ}C$, respectively. The pH and temperature stabilities of the recombinant enzyme were significantly enhanced and the half life at 55$^{\circ}C$ was about 60 hr. When the recombinant CGTase was reacted with 5% soluble starch, the conversion yield of total cyclodextrin (CD) from starch was estimated to be 41% at 48 hr, whereas the wild type enzyme showed the yield of 12%. This improvement of conversion yield and thermal stability of CGTase may be useful for the development of low-cost CD production process.

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수성2상계를 이용한 Cyclodextrin Glucanotransferase 분리 및 회수 (Separation and Recovery of Cyclodextrin Glucanotransferase Using Aqueous Two-Phase Systems)

  • 김진현;홍승서;이현수
    • KSBB Journal
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    • 제15권6호
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    • pp.556-559
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    • 2000
  • Cyclodextrin Glucanotransferase(EC 2.4.1.19 : 1,4-${\alpha}$-glucano) transferase, cyclizing; CGTase) can be separated and recovered in an aqueous two-phase system composed of poly(ethylene glycol)(PEG)/dextran and PEG/salt. In an aqueous two-phase system consisting of PEG 35000 (5%) and dextran T2000 (7%), all cell and debris were collected at the interphase. CGTase partitioned to the denser dextran phase at an yield of 83.4%. On the other hand, in an aqueous two-phase system consisting of PEG 35000 (10%) and sodium phosphate (15%), CGTase partitioned to the denser salt phase at an yield of 95.5%. In order to recover CGTase using an aqueous two-phase system, the PEG/salt system proved to be more efficient than the PEG/dextran system in terms of yield and cost.

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Cyclodextrin Glucanotransferase 고생산 호알칼리성 세균의 탐색과 분비 효소의 특성 (Screening of Alkalophilic Bacillus sp. for Overproduction of Cyclodextrin Glucanotransferase and Its Enzymatic Properties)

  • 도은주;박종부;이용현
    • 한국미생물·생명공학회지
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    • 제21권2호
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    • pp.119-124
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    • 1993
  • An alkalophilic microorganism for overproduction of cyclodextrin glucanotransferase (CGTase) was newly isolated from hot-water spring soil, and identified as Bacillus firmus var. alkalophilus H609. The strain maintained stability during preservation and cultivation for the enzyme production, and produced significant amount of CGTase corresponding to the volumetric activity of 75 units/mL at 37C, initial pH of 11.2, and after 40 hours. The strain excreted several different proteins showing CGTase activity that catalyzed the formation of mainly beta-and Gamma-type cyclodextrin (ratio of 7:1) from soluble starch without accumulation of alpha-type. Other enzymatic properties were also investigated.

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김치에서 분리한 Bacillus sp. S-6의 Cyclodextrin Glucanotransferase의 특성과 최적생산조건 (Some Properties and Optimal Culture Conditions of Cyclodextrin Glucanotransferase of Bacillus sp. S-6 Isolated from Kimchi)

  • 전홍기;조영배;김수진;배경미
    • 한국식품영양과학회지
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    • 제27권4호
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    • pp.609-617
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    • 1998
  • A microorganism capable of producing high level of extracellular cyclodextrin glucanotransferase(EC 2.4.1.19 ; CGTase) was isolated from Kimchi. 2-O-$\alpha$-D-glucopyranosyl L-ascorbic acid(AA-2G) was synthesized by transglycosylation reaction of CGTase using starch as a donor and L-ascorbic acid as an acceptor. The isolated strain S-6 was identified as Bacillus sp. S-6. The maximal CGTase production was observed in a medium containing 0.5% soluble starch, 1% yeast extract, 1% NaCO3, 0.1% K2HPO4, and 0.02% MgSO4 with initial pH 8.0. The strain was cultured at 37$^{\circ}C$ for 40 hr with reciprocal shaking. Using the culture supernatant as crude enzyme, the optimal pH and temperature of the CGTase activity of this strain were 7.0 and 4$0^{\circ}C$. In the effects of pH and temperature on the stability of the enzyme, the enzyme was stable in the range of pH 6.0~10.0 and up to 45$^{\circ}C$, respectively.

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