• Title/Summary/Keyword: cyclodextrin glucanotransferase

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

  • 김진현;홍승서;이현수
    • KSBB Journal
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    • v.16 no.2
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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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Purification and Characterization of Cyclodextrin Glucanotransferase from Bacillus stearothermophilus KJ16 (Bacillus stearothermophilus KJ16이 생산하는 Cyclodextrin Glucanotransferase 의 정제와 효소특성)

  • Kwon, Hyun-Ju;Nam, Soo-Wan;Kim, Kwang-Hyun;Song, Seong-Koo;Yun, Jong-Won;Kim, Byung-Woo
    • Journal of Life Science
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    • v.8 no.3
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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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Purification of Cyclodextrin Glucanotransferase by Affinity Chromatography (Affinity Chromatography를 이용한 Cyclodextrin Glucanotransferase의 정제)

  • 안중훈;황진봉;김승호;김경은
    • Microbiology and Biotechnology Letters
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    • v.19 no.3
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    • pp.313-314
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    • 1991
  • - The cyclodextrin glucanotransferase (CGTase) of a mutant of Bacillus stearothermophilus was purified in one step by affinity chromatography. The recovery was 95%. The specific activity of the CGTase increased from 26.2 U/mg protein to 485.5 U/mg protein. The purified CGTase was almost homogeneous by SDS-polyacrylamide gel electrophoresis. The one-step purification proved to be feasible with the mutant in contrast to the parent strain, which required pre-purification step of ammonium sulfate precipitation.

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

  • 안중훈;황진봉;김승호
    • Microbiology and Biotechnology Letters
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    • v.19 no.4
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    • pp.368-371
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    • 1991
  • The influence of ethylene glycol, glycerol, sorbitol and sucrose on the thermostability of Bacilus stearothermophzlus cyclodextrin glucanotransferase (CGTase) was investigated. Glycerol, sorbitol and sucrose had effect on thermostability of the CGTase. The effects appeared to be strongly dependent on concentration of additives. The thermostability of CGTase also was assayed in organic solvents such as n-butanol, l, &dioxane, n-octane. The therrnostability of CGTase increased in l, 4-dioxane and n-octane. Particularly, in n-octane, the CGTase retained the 81% of the initial activity after incubation at $75^{\circ}C$ for 90 min.

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

  • 안중훈;황진봉;김승호
    • Microbiology and Biotechnology Letters
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    • v.18 no.6
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    • pp.585-590
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    • 1990
  • The cyclodextrin glucanotransferase (CGTase) was purified from the culture broth of Bacillus stearothermophilus by ammonium sulfate precipitation and affinity chromatography. The specific activity of the CGTase increased by about 31-fold from 111.5 U/mg protein to 3445.0 Ulmg protein. The SDSPAGE indicated that the purified CGTase was homogeneous and the molecular weight of the purified CGTase was about 78,000. The optimum pH and temperature was 6.0 and $60^{\circ}C$, respectively. This enzyme was stable from pH 5.5-10.0. The enzyme retained its full activity at the incubation temperature up to $60^{\circ}C$ and calcium ion increased the thermal stability. The isoelectric point was about 4.8.

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

  • 이용현;백승걸;박동찬;신현동
    • Microbiology and Biotechnology Letters
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    • v.21 no.5
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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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Biochemical Properties of Recombinant Cyclodextrin Glucanotransferase Expressed in Saccharomyces cerevisiae (Saccharomyces cerevisiae에 발현된 재조합 cyclodextrin glucanotransferase의 생화학적 특성)

  • 박현이;남수완;김병우
    • Journal of Life Science
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    • v.11 no.3
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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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Separation and Recovery of Cyclodextrin Glucanotransferase Using Aqueous Two-Phase Systems (수성2상계를 이용한 Cyclodextrin Glucanotransferase 분리 및 회수)

  • 김진현;홍승서;이현수
    • KSBB Journal
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    • v.15 no.6
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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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Screening of Alkalophilic Bacillus sp. for Overproduction of Cyclodextrin Glucanotransferase and Its Enzymatic Properties (Cyclodextrin Glucanotransferase 고생산 호알칼리성 세균의 탐색과 분비 효소의 특성)

  • 도은주;박종부;이용현
    • Microbiology and Biotechnology Letters
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    • v.21 no.2
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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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Some Properties and Optimal Culture Conditions of Cyclodextrin Glucanotransferase of Bacillus sp. S-6 Isolated from Kimchi (김치에서 분리한 Bacillus sp. S-6의 Cyclodextrin Glucanotransferase의 특성과 최적생산조건)

  • 전홍기;조영배;김수진;배경미
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.4
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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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