• KSBB Journal
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• 제24권6호
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• pp.570-578
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• 2009

헤마토코커스로부터 아스타잔틴의 추출과 제형화의 유효성을 확인하고 적용성을 검토하고자 헤마토코커스 추출물과 아스타잔틴 포접화합물의 생리활성을 측정하였다. 그 결과, 헤마토코커스 추출물은 DPPH 라디칼 소거능, xanthine oxidase 저해 활성 및 hydroxyl 라디칼 소거능에 대해 in vitro 실험에서 활성을 나타냈다. 또한 헤마토코커스 추출물은 대조군인 kojic acid에 비해 동일한 농도에서 2배 이상의 미백활성을 나타냈고, 자궁암 세포주 (HeLa)에 대해 대조항암물질인 5-fluorouracil (5-FU)과 비슷한 항암활성을 나타냈다. 아스타잔틴은 또한 숙취해소 효과를 갖고 있는 것으로 확인되었는데, 대조군인 지구자 추출물에 비해 1.5배 높음을 알 수 있었다. 또한, 아스타잔틴과 싸이클로덱스트린의 포접화합물 As-$\beta$-CD와 헤마토코커스 추출물과 싸이클로 덱스트린의 포접화합물인 H.p.-$\beta$-CD의 생리활성을 조사한 결과, As-$\beta$-CD는 대조군인 지구자추출물보다 높은 숙취해소 효과를 나타냈고, H.p.-$\beta$-CD는 대조군인 kojic acid와 비슷한 수준의 미백효과를 나타냈다.

• 생명과학회지
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• 제8권5호
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• pp.497-503
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• 1998

GTase와 CDase를 함께 분비$\cdot$생산하는 B. stearother-mophilus KJl6 균주의 CDase를 ammonium sulfate 침전, DBAE-cellulose, Sephadex G-100 column chromatogra-phy, 및 FPLC로 수율 7%, 비활성 12.4 units/mg, 정제도 87.6배로 정제된 CDase를 얻었으며 SDS-PAGE 상 단일 band를 확인하였다. 정제된 CDase의 분자량은 약 68,000 dalton 이었고 활성 최적 pH와 온도는 6.0와 55$^{\circ}C$였다. pH 안정성은 5.5~8.5의 범위에서 비교적 안정하였으며, 온도 안정성은 5$0^{\circ}C$에서 2시간까지는 안정하였고, 7$0^{\circ}C$에서 1시간 전처리하여도 80% 이상의 잔존활성을 나타내었다. 효소 활성은 $Cu^{+2}$와 $Hg^{+2}$와 같은 금속이온과 p-chlorome-rcuribenzoate, N-bromosuccinimide, mercaptoethanol, dithiothreitol에 의해서 효소활성이 강하게 저해되었다. 기질에 대한 반응 특이성은 $\gamma$ -CD를 가장 잘 분해하였으며, 그 외에 soluble starch나 amylose, amylopectin 등의 기질도 잘 분해하나 이들의 분해속도는 $\gamma$-CD에 비해서는 늦었다. 이들 기질의 최종 분해산물은 maltose였으며, maltose는 거의 분해되지 않았다.

• Journal of Microbiology and Biotechnology
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• 제2권2호
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• pp.85-91
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• 1992

The intestinal microflora of humans is an extraordinarily complex mixture of microorganisms, the majority of which are anaerobic microorganisms. The distribution of amylolytic microorganisms in the human large intestinal tract was investigated in various individuals of differing ages using anaerobic culture techniques. A large percentage of the amylolytic microorganisms present belonged to the Genus Bifidobacteria. The number of Bifidobacteria increased significantly at two years of age. Adults and children above 2 years old carried about $0.8{\times}10^9-2.0{\times}10^{10}$ colony forming units (CFU/gram) of amylolytic Bifidobacteria. Among these amylolytic Bifidobacteria, Int-57 was chosen for further studies. Between 65% and 85% of the amylase produced was secreted and the remaining amylase was bound to the cell wall facing the outside. Amylase production could be induced by starch in a stable form. When cells were grown on maltose or glucose, amylase production was much lower than on starch and amylase activity disappeared after 24 hours growth on these media. Partially purified enzymes showed optimum activity at a temperature of $50^{\circ}C$ and at an optimum pH of 5.5, respectively. Heat treatment at $70^{\circ}C$ for 30 minutes almost completely inactivated amylase. The hydrolysis products of starch were mainly maltose and maltotriose. Soluble starch, amylose, amylopectin, and $\gamma$-cyclodextrin($\gamma$-CD) were easily hydrolyzed. The rate of hydrolysis of $\alpha$-CD and $\beta$-CD was slower than that of $\gamma$-CD. Carboxymethyl cellulose, $\beta$-1, 3-glucan and inulin were not hydrolyzed.

• BMB Reports
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• 제37권4호
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• pp.408-415
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• 2004

The expression of the Paenibacillus sp. A11 cyclodextrinase (CDase) gene using the pUC 18 vector in Escherichia coli JM 109 resulted in the formation of an insoluble CDase protein in the cell debris in addition to a soluble CDase protein in the cytoplasm. Unlike the expression in Paenibacillus sp. A11, CDase was primarily observed in cytoplasm. However, by adding 0.5 M sorbitol as an osmolyte, the formation of insoluble CDase was prevented while a three-fold increase in cytoplasmic CDase activity was achieved after a 24 h-induction. The recombinant CDase protein was purified to approximately 14-fold with a 31% recovery to a specific activity of 141 units/mg protein by 40-60% ammonium sulfate precipitation, DEAE-Toyopearl 650 M, and Phenyl Sepharose CL-4B chromatography. It was homogeneous by non-denaturing and SDS-PAGE. The enzyme was a single polypeptide with a molecular weight of 80 kDa, as determined by gel filtration and SDS-PAGE. It showed the highest activity at pH 7.0 and $40^{\circ}C$. The catalytic efficiency ($k_{cat}/K_m$) values for $\alpha$-, $\beta$-, and $\gamma$-CD were $3.0{\times}10^5$, $8.8{\times}10^5$, and $5.5{\times}10^5\;M^{-1}\;min^{-1}$, respectively. The enzyme hydrolyzed CDs and linear maltooligosaccharides to yield maltose and glucose with less amounts of maltotriose and maltotetraose. The rates of hydrolysis for polysaccharides, soluble starch, and pullulan were very low. The cloned CDase was strongly inactivated by N-bromosuccinimide and diethylpyrocarbonate, but activated by dithiothreitol. A comparison of the biochemical properties of the CDases from Paenibacillus sp. A11 and E. coli transformant (pJK 555) indicates that they were almost identical.

• Journal of Pharmaceutical Investigation
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• 제31권3호
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• pp.151-160
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• 2001

The purpose of this study is to investigate the interaction of progesterone with various cyclodextrins (CDs) in the aqueous solution and in solid state, and finally to formulate a parenteral aqueous formulation. CDs used were ${\alpha}-$, ${\beta}-$, and ${\gamma}-CD$, $2-hydroxypropyl-{\beta}-CD$ (HPCD), sulfobutyl $ether-{\beta}-CD$ (SBCD), $dimethyl-{\beta}-CD$ (DMCD) and $trimethyl-{\beta}-CD$ (TMCD). The solubility studies of progesterone were performed in the presence of various CDs as a function of concentration or temperature. The solubility of progesterone increased in the rank order of ${\alpha}-CD$ < ${\beta}-CD$ < ${\gamma}-CD$ < TMCD$ < HPCD < DMCD < SBCD. Addition of SBCD (200 mg/ml) in water increased the aqueous solubility $(9.36\;{\mu}g/ml)$ about 3,200 times, and lowering the temperature facilitated the solubilization of progesterone. However, the addition of HPCD and SBCD in 20:80 (v/v) polyethylene glycol 300-water and propylene glycol-water cosolvents markedly decreased the solubility of progesterone, compared with solubilizing effects in water. Physical mixtures and solid dispersions of progesterone with HPCD or SBCD were prepared, and evaluated by differential scanning calorimetry (DSC), Fourier-transform infrared spectroscopy (FT-IR), near IR spectroscopy and dissolution studies. By DSC and IR studies, it was found that progesterone was dispersed in HPCD in monotectic state and dissolved rapidly from both solid dispersions. Based on solubility studies, new aqueous progesterone fonnulations (5 mg/ml) containing SBCD (200 mg/ml) could be prepared and did not form precipitates even after 2 months at $4^{\circ}C$. The solution was transparent when mixed with normal saline and 5% dextrose injection at 1: 1, 1:10 and 1:20 (v/v) even after 7 days. Permeation rates of progesterone through a cellulose membrane from 20% PEG 300 solution $(50\;{\mu}g/ml)$ containing HPCD or SBCD were compared with oily formulation. Permeation of progesterone from oily formulation did not occur up to 8 hr, but aqueous formulations showed fast permeation rates from early stage of permeation study. The addition of HPCD or SBCD retarded the permeation rates of progesterone with the increase of CD concentrations, suggesting the possibility of a controlled absorption from the site administered intramuscularly. These results demonstrate that it is feasible to develop a new progesterone parenteral aqueous injection (5 mg/ml) using SBCD.