• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.2
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• pp.351-357
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• 1997

The cyclodextrin glycosyltransferase(EC 3.2.1.19) from Bacillus firmus was purified by precipitating with ammonium sulfate followed by, DEAE-Sephadex A-50 column chromatography and Sephadex G-100 column chromatography. In this way, we were able to obtain the single band protein on SDS-PAGE with a yield of 12%, whose purity was 49 fold. The purified CGTase was identified as a protein having molecular weight of approximately 80,000 dalton and isoelectric point of 9.6. The optimum pH and temperature for the enzyme activity were 8.0 and $65^{\circ}C$, respectively. The enzyme was stable at between pH 5.5 and 9.0 and up to $50^{\circ}C$. After 24hr of enzyme reaction using soluble starch as substrate, the ratio of ${\alpha}-$, ${\beta}-$ and ${\gamma}-cyclodextrin$ production was 0.01 : 2.90 : 1.00, respectively. And this CGTase pro-duced mainly ${\beta}-$ and ${\gamma}-cyclodextrin$.

• Journal of Life Science
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• v.8 no.5
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• pp.614-621
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• 1998

An extracellular inulinase from Penicillium sp. which isolated from soil sample was purified to a single protein th-rough ammonium sulfate fractionation, DEAE-Sephacel ion exchange chromatography and Toyopearl HW 65 F gel filtration. The temperature and pH for the enzyme reaction were around 6$0^{\circ}C$ and 4.0, respectively. The enzyme was stable at 3$0^{\circ}C$-5$0^{\circ}C$ and in the pH range of 4 to 5. $CuCl_2$, $HgCl_2$ and EDTA inhibited the enzyme activity strongly. By contrast, $MnCl_2$ and $CaCl_2$ activated the enzyme activity. The molecular weight of the purified enzyme was esti-mated to be 77,000 dalton by SDS-polyacrylamide gel electrophoresis. The Km values of the enzyme for inulin were calculated to be $2.2\times10^{-3}$M. TLC analysis suggested that purified enzyme is exo-type inulinase. The NH2-terminal amino acid sequences of the purified enzyme was determined to be $NH_2$-X-Glu-Ser-Tyr-Thr-Glu-Lys/Leu-Tyr-Arg-Pro.

• Microbiology and Biotechnology Letters
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• v.16 no.2
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• pp.92-97
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• 1988

Cysteinylglycinase was partially purified from Proteus mirabilis by consecutive procedure. The specific activity was increased about 16-fold to that of cell-free extract. The enzyme was found rather unstable on ammonium sulfate precipitation ann the precipitated enzyme protein became partially insoluble during dialysis. The precipitated enzyme was found to be solubilized by treatment of 4% Triton X-100 effectiviely, The optimum temperature and pH of the enzyme activity were 35$^{\circ}C$ and 7.3, respectively. After heat treatment of the enzyme at 5$0^{\circ}C$ for 30 min, it lost the activity to 70%. The enzyme was stable at pH 7.0-8.0. The molecular weight of the cysteinylglycinase was found to be about 190,000 by Sephadex G-150 gel filtration. The enzyme was activated by the addition of Mn$^{2+}$ and $Mg^{2+}$ ions. The maximal activation was obtained in preincubation with $Mg^{2+}$ ion for 30 min. The enzyme catalyzed the hydrolysis of various dipeptides and tripeptides. The Km and Vmax values for cysteinylglycine were 1.60 mM and 0.24 m unit/ mg, respectively.

• Microbiology and Biotechnology Letters
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• v.30 no.4
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• pp.293-297
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• 2002

The cyclodextrin glucanotransferase (CGTase) gene from Bacillus stearothermophilus NO2 was expressed in Saccharomyces cerevisiae 2805 under the adhl promoter. The CGTase was purified from S. cerevisiae 2805/pVT-CGTS. The purified enzyme exhibited a optima of activity around pH 7.0 and $65^{\circ}C$. Thermal stability of the enzyme was increased fairly as compared with the CGTase of B. stearothermophilus NO2. The conversion yield of cyclodextrin (CD) and the production ratio of $\alpha$-, $\beta$,-, ${\gamma}$-CD from starch were showed similarly aspect to the CGTase of B. stearothermophilus NO2.

• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.559-564
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• 1992

To increase the efficency of utilizing cellulosic biomass, a potent xylanase producing bacteria was isolated and identified as Bacillus sp. N-25. Extracellular xylanase from Bacillus sp. N-25 was partially purified by ammonium sulfate precipitation, DEAE-Sephadex A-25 and Sephadex G-IOO column chromatographies. The xylanase was single fraction on chromatography and was true xylanase without cellulase activity. The enzyme was stable at pH 6-8 and 80% activity was remained at $50^{\circ}C$ for 30 min, but was inhibited by $Hg^{2+}$, $Ag^{2+}$, and $Mn^{2+}$. From the fact that the major end product was xylose, we suggested that the enzyme is an exo-xylanase which may be a prime candidate for industrial use.

• Microbiology and Biotechnology Letters
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• v.17 no.3
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• pp.198-201
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• 1989

Steptomyces sp. GI 32 with high production of glucose isomerase was isolated from soil. The maximum enzyme production was observed in the culture medium containing 1% sorbitol, 0.6% tryptone, 0.4% yeast extract, 1mM Fe$_2$(SO$_4$)$_3$ with initial pH 7.0 when the cell was cultured at 35$^{\circ}C$ about 18 hours with shaking. The enzyme was partially purified by ammonium sulfate fractionation and DEAE cellulose chromatography. The enzyme was also appeared to be relatively thermostable, and no apopreciable inactivation was observed after incubation at 7$0^{\circ}C$ for 1 hour. The optimal pH and temperature of the enzyme were pH 8.0 and 7$0^{\circ}C$, respectively.

• YAKHAK HOEJI
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• v.51 no.3
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• pp.199-205
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• 2007

Thermostable asparaginase was purified to homogeneity from mesophilic Strepfomyces lincolnensis M-20 by 30${\sim}$70% ammonium sulfate precipitation and asparagine-Sepharose CL 6B affinity column chromatography, The apparent molecular mass of L-asparaginase by SDS-PAGE was found to be 47 kDa, whereas by its mobility on Sephacryl S-300 column was around 180 kDa, indicating that the enzyme at the native stage acts as tetramer, The purified enzyme showed a single band on acrylamide gel electrophoresis. The optimum pH and temperature were pH 9.5 and 55${\circ}$C, respectively. Chemical modification experiments of purified asparagines implied the existence cystein residue located at or near active site. Purified asparaginase retained the 85% of the initial activity after incubation at 90${\circ}$C for 30 min. A correlation between themostability and resistance to proteolysis of commercial asparaginase and purified asparaginase from Strepfomyces lincolnensis M-20 was investigated. Purified thermostable asparaginase was resistant to trypsin and chymotrypsin treatment, while the commercial asparaginase was not themostable and was susceptible to proteolytic treatment with trypsin and chymotrypsin.

• Proceedings of the Korean Society of Propulsion Engineers Conference
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• 2017.05a
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• pp.409-411
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• 2017

미사일 추력기 체계에 적용되는 하이드라진[$N_2H_4$]추진제는 MSDS-OHS 유해성 분류상 급성독성 물질로서 사용이 제한되고 있는 바, 다양한 대체물질이 개발 중이다. 최근 해외에서 안전성과 취급이 우수한 질산 히드록실암모늄[$NH_3OHNO_3$]과 암모늄 디나이트라마이드[$NH_4N(NO_2)_2$] 기반 단일계 액상추진제가 개발중이며, 이 물질들을 이용한 추력기 시스템 적용 시험이 진행되고 있다. 그러나 저온에서의 연로물질 산성화 반응으로 인한 디나이트라마이드[$N(NO_2)_2{^-}$] 물질의 분해는 나이트레이트[$NO_3{^-}$] 이온 생성을 촉진시키며, 부수적으로 발생하는 침전물은 촉매 및 노즐의 막힘 현상을 유발하므로 추력기 성능의 저해요인으로 작용한다. 그러므로 저온분해 방지를 위한 첨가제 조성 개발 및 열분해 특성 연구가 최근의 관심사이다. 본 연구는 합성/정제/추출한 암모늄 디나이트라마이드 산화제를 주요 조성물로 적용하였으며, 염기성 안정화제를 질량비율 4~5% 첨가하여 산성화 반응을 억제시킨 단일계 액상추진제(KMP) 형태로 제조하였다. 합성한 추진제는 시차주사열량계(DSC)를 이용하여 분해온도를 측정하여 열안정성을 평가해보았다.

• Proceedings of the KAIS Fall Conference
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• 2004.06a
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• pp.303-306
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• 2004

참멍게의 체액세포로부터 산추출 후, 조 추출물에서 천연항균소재를 개발하기 위해 먼저 멍게 조 추출액을 직접 Tricine-SDS PAGE를 통하여 주요 펩타이드들의 분자량의 범위를 살펴본 결과 6kDa 이하의 분자량의 펩타이드들이 다량 존재함을 알수 있었다. 펩타이드들의 size별 항균활성을 알아보기 위해 여러 사이즈의(100, 50, 30, 10 kDa)의 한외여과만으로 여과하여 그 여과액들의 specific 활성을 알아본 결과 여과막의 cut-off size에 상관없이 거의 인정한 specific activity를 가짐을 알 수 있었다. 멍게 조 추출액의 여러 미생물에 대한 항균 스펙트럼을 알아보기 위해 E.coli, P. aeruginosa, S. typhi, V. parahaemolyticus, L. monocytogenes, B. sutillus, S. aureus, S. mutans 균주들을 $10^5 CFU/ml$로 부터 4log 감소시키는 농도를 측정한 결과 각각 200, 50, 60, 10, 25, 30, 100, 100ppm 농도였으며, 대표적 상용화 항균 펩타이드인 Nisin과의 항균활성 비교 결과 비슷하거나 월등히 뛰어난 결과를 보여주었다. 또한 추출액의 열안정성을 측정하기 위해 $100^{\circ}C$에서 10분간 가열한 후 원액과의 항균력의 차이를 Radial diffusion assay로 알아본 결과 항균력의 차이가 거의 없음을 알 수 있었다.

• The Korean Journal of Mycology
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• v.24 no.4 s.79
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• pp.293-304
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• 1996

A bacterial strain, KSl, possessing strong antifungal activity was isolated from soil samples of ginseng fields and identified as Bacillus subtilis. In greenhouse test, the culture filtrate of B. subtilis KS1 showed strong protective effect against several fungal diseases of agricultural plants such as cucumber gray mold and wheat leaf rust. In addition, the crude butanol fraction of the culture filtrate exhibited antagonistic effect against several fungi including plant or human pathogens, such as Botrytis maydis, Chytridium lagenarium and Candida albicans. The antifungal compound, SW1, produced by B. subtilis KS1 was purified through consecutive chromatographic separations on a pep-RPC column and a ${\mu}$ Bondapak $C_{18}$ reverse phase column. Temperature and pH showed little effect on the stability of the compound in the ranges $-20-121^{\circ}C$ and pH 4.0-10.0, respectively. The composition and structural characteristics of SW1 were analysed by HPLC and by $^1H-,\;^1H-^1H-COSY$, NOESY, COSY-NOESY and HOHAHA NMR spectroscopy, respectively, which revealed that the compound belongs to iturin A, a typical cyclic antifungal compound produced by B. subtilis. In contrast to the previously reported iturin A compounds which have one or no $-CH_3$ side chain in the hydrophobic hydrocarbon chain of ${\beta}-amino$ acids, SW1 was shown to have a ${\beta}-amino$ acid containing 12-carbon skeleton with two $-CH_3$ side chains.