• BMB Reports
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• 제38권2호
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• pp.198-205
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• 2005

Resveratrol (RES) and genistein (GEN) are the dietary natural products known to possess chemopreventive property and also the ability to repair DNA damage induced by mutagens/carcinogens. It is believed that the therapeutic activity of these compounds could be primarily due to their interaction with nucleic acids but detailed reports are not available. We here explore the interaction of these drugs with nucleic acids considering DNA and RNA as a potential therapeutic target. The interaction of RES and GEN has been analysed in buffered solution with DNA [saline sodium citrate (SSC)] and RNA [tris ethylene diammine tetra acetic acid (TE)] using UV-absorption and Fourier transform infrared (FTIR) spectroscopy. The UV analysis revealed lesser binding affinity with nucleic acids at lower concentration of RES (P/D = 5.00 and 10.00), while at higher drug concentration (P/D = 0.75, 1.00 and 2.50) hyperchromic effect with shift in the ${\lambda}_{max}$ is noted for DNA and RNA. A major RES-nucleic acids complexes was observed through base pairs and phosphate backbone groups with K = $35.782\;M^{-1}$ and K = $34.25\;M^{-1}$ for DNA-RES and RNA-RES complexes respectively. At various concentrations of GEN (P/D = 0.25, 0.50, 0.75, 1.00 and 2.50) hyperchromicity with shift in the ${\lambda}_{max}$ from 260 $\rightarrow$ 263 om and 260 $\rightarrow$ 270 nm is observed for DNA-GEN and RNA-GEN complexes respectively. The binding constant (from UV analysis) for GEN-nucleic acids complexes could not be obtained due to GEN absorbance overlap with that of nucleic acids at 260 nm. Nevertheless a detailed analysis with regard to the interaction of these drugs (RES/GEN) with DNA and RNA could feasibly be understood by FTIR spectroscopy. The NH band of free DNA and RNA which appeared at $3550-3100\;cm^{-1}$ and $3650-2700\;cm^{-1}$ shifted to $3450-2950\;cm^{-1}$ and $3550-3000\;cm^{-1}$ in DNA-RES and RNA-RES complexes respectively. Similarly shifts corresponding to $3650-3100\;cm^{-1}$ and $3420-3000\;cm^{-1}$ have been observed in DNA-GEN and RNA-GEN complexes respectively. The observed reduction in NH band of free nucleic acids upon complexation of these drugs is an indication of the involvement of the hydroxyl (OH) and imino (NH) group during the interaction of the drugs and nucleic acids (DNA/RNA) through H-bonded formation. The interaction of RES and GEN with bases appears in the order of G $\geq$ T > C > A and A > C $\geq$ T > G. Further interaction of these natural compounds with DNA and RNA is also supported by changes in the vibrational frequency (shift/intensity) in symmetrical and asymmetrical stretching of aromatic rings of drugs in the complex spectra. No appreciable shift is observed in the DNA and RNA marker bands, indicating that the B-DNA form and A-family conformation of RNA are not altered during their interaction with RES and GEN.

• Journal of Microbiology and Biotechnology
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• 제26권10호
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• pp.1661-1667
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• 2016

The ginsenoside-hydrolyzing β-glucosidase gene (bgy2) was cloned from Lactobacillus brevis. We expressed this gene in Escherichia coli BL21(DE3), isolated the resulting protein, and then utilized the enzyme for the biotransformation of ginsenosides. The bgy2 gene contains 2,223 bp, and encodes a protein of 741 amino acids that is a member of glycosyl hydrolase family 3. β-Glucosidase (Bgy2) cleaved the outer glucose moieties of ginsenosides at the C-20 position, and the inner glucose at the C-3 position. Under optimal conditions (pH 7.0, 30℃), we used 0.1 mg/ml Bgy2 in 20 mM sodium phosphate buffer (PBS) for enzymatic studies. In these conditions, 1.0 mg/ml ginsenoside Rb1 and ginsenoside F2 were converted into 0.59 mg/ml ginsenoside Rd and 0.72mg/ml compound K, with molar conversion productivities of 69% and 91%, respectively. In pharmaceutical and commercial industries, this recombinant Bgy2 would be suitable for producting ginsenoside Rd and compound K.

• The Korean Journal of Physiology and Pharmacology
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• 제3권4호
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• pp.447-454
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• 1999

The production of nitric oxide (NO) and the expression of inducible nitric oxide synthase (iNOS) are known to be modulated by a variety of factors. Recent study showed that endotoxin-induced NO synthesis and iNOS expression were greatly enhanced by elevation of extracellular glucose concentration in murine macrophages. Although this was suggested to be due to the activation of protein kinase C (PKC) via sorbitol pathway, there was lack of evidence for this speculation. This study was performed to delineate the underlying intracellular mechanisms of glucose-enhancing effect on endotoxin-induced NO production in Raw264.7 macrophages. The levels of NO release induced by lipopolysaccharide (LPS) significantly increased by the treatment of glucose in a concentration dependent manner and also, this effect was observed in LPS-preprimed cells. Concurrent incubation of cells with PKC inhibitors, H-7 or chelerythrine, and LPS resulted in the diminution of NO production regardless of glucose concentration but this was not in the case of LPS-prepriming, that is, chelerythrine showed a minimal effect on the glucose- enhancing effect. PMA, a PKC activator, did not show any significant effect on glucose-associated NO production. Modulation of sorbitol pathway with zopolrestat, an aldose reductase inhibitor, did not affect LPS-induced NO production and iNOS expression under high glucose condition. And also, sodium pyruvate, which is expected to normalize cytosolic $NADH/NAD^+$ ratio, did not show any significant effect at concentrations of up to 10 mM. Glucosamine marginally increased the endotoxin-induced nitrite release in both control and high glucose treated group. 6-diazo-5-oxonorleucine (L-DON) and azaserine, glutamine: fructose- 6-phosphate amidotransferase (GFAT) inhibitors, significantly diminished the augmentation effect of high glucose on endotoxin-induced NO production. On the other hand, negative modulation of GFAT inhibitors was not reversed by the treatment of glucosamine, suggesting the minimal involvement, if any, of glucosamine pathway in glucose-enhancing effect. In summary, these results strongly suggest that the hexosamine biosynthesis pathway and the activation of PKC via sorbitol pathway do not contribute to the augmenting effect of high glucose on endotoxin induced NO production in macrophage-like Raw264.7 cells.

• 한국산학기술학회논문지
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• 제15권9호
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• pp.5644-5651
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• 2014

본 연구는 칸디다 알비칸스(C. albicans)에 대한 프로폴리스의 억제효과를 확인하고자 수행되었다. 프로폴리스는 양봉 꿀벌에서 채취하여 C. albicans는 $37^{\circ}C$에서 액체 배지에서 2시간 배양 하였다. 항균 활성검삼를 위해서 생리식염수 (PBS), 3% 차아염소산나트륨 (NaOCl에), 0.1% 클로로헥시딘 (CHX), 프로 폴리스 추출물 ($5{\mu}L/m{\ell}$, $10{\mu}L/m{\ell}$)에서 평가하였다. C. albicans는 3%의 NaOCl, 0.1% CHX, 프로 폴리스($5{\mu}L/m{\ell}$, $10{\mu}{\ell}/m{\ell}$)에서 15, 14.5, 16, 17mm의 억제구간을 확인하였다. 항균활성을 확인하기 위하여 집락형성을 분석한 결과, 3%의 NaOCl, 0.1 % CHX, $5{\mu}L/m{\ell}$와 $10{\mu}L/m{\ell}$의 프로 폴리스가 7, 7, 5, 7 로그지수가 감소를 보였다. C. albicans서는 $10{\mu}L/m{\ell}$의 프로폴리스에서 유의하게 억제되는 결과를 보여주었다. 이러한 결과를 통하여 프로폴리스는 구강 점막 질환에서 새로운 향균 약제가 될수 있는 것으로 생각된다.

• Applied Biological Chemistry
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• 제43권4호
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• pp.247-252
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• 2000

한국 전통장류의 기능성 탐색 연구의 일환으로 청국장 발효과정 중 고혈압을 유발하는 angiotensin converting enzyme(ACE)의 저해 peptide를 분리하고 저해효과를 검토하였다. 청국장은 Bacillus subtilis CH-1023을 이용하여 제조하였고, $20^{\circ}C,\;30^{\circ}C,\;40^{\circ}C,\;50^{\circ}C,\;60^{\circ}C$에서 $0{\sim}72$시간동안 배양하면서 protein양, protease activity, ACE 저해율을 측정하고 저해활성을 가지는 peptide를 정제 후 아미노산조성을 분석하였다. 청국장의 발효시간이 경과함에 따라 protein 함량이 증가하여 $40^{\circ}C$, 60시간에서 최대를 나타낸 후 감소하였으며, 발효시간에 따른 protease activity와 ACE 저해율은 $40^{\circ}C$, 60시간에서 최대로 나타난 후 점차 감소하였다. 따라서 Bacillus subtilis CH-1023을 이용한 청국장 제조는 $40^{\circ}C$에서 60시간 배양이 최적의 배양조건이었다. 최적 발효조건에 따라 청국장을 제조한 후 20 mM sodium phosphate buffer(pH 7.0)를 가해 추출한 추출물을 Amicon YM-3 membrane filtration과 Sephadex G-10, G-25를 이용한 gel filtration으로 부분정제 하였다. 또한 정제한 peptide는 첨가함량이 높아질수록 저해활성은 높게 나타났으며, 0.5 mg 정도의 peptide 함량으로 94.3%의 저해율을 나타내었다. 정제한 peptide의 아미노산 조성은 histidine, alanine, phenylalanine 함량이 높은 것으로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제28권4호
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• pp.606-612
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• 2018

The enzyme xylose isomerase (E.C. 5.3.1.5, XI) is responsible for the conversion of an aldose to ketose, especially xylose to xylulose. Owing to the ability of XI to isomerize glucose to fructose, this enzyme is used in the food industry to prepare high-fructose corn syrup. Therefore, we studied the characteristics of XI from Anoxybacillus kamchatkensis G10, a thermophilic bacterium. First, the gene coding for XI (xylA) was inserted into the pET-21a(+) expression vector and the construct was transformed into the Escherichia coli competent cell BL21 (DE3). The expression of recombinant XI was induced in the absence of isopropyl-thio-${\beta}$-galactopyranoside and purified using Ni-NTA affinity chromatography. The optimum temperature of recombinant XI was $80^{\circ}C$ and measurement of the heat stability indicated that 55% of residual activity was maintained after 2 h incubation at $60^{\circ}C$. The optimum pH was found to be 7.5 in sodium phosphate buffer. Magnesium, manganese, and cobalt ions were found to increase the enzyme activity; manganese was the most effective. Additionally, recombinant XI was resistant to the presence of $Ca^{2+}$ and $Zn^{2+}$ ions. The kinetic properties, $K_m$ and $V_{max}$, were calculated as 81.44 mM and $2.237{\mu}mol/min/mg$, respectively. Through redundancy analysis, XI of A. kamchatkensis G10 was classified into a family containing type II XIs produced by the genera Geobacillus, Bacillus, and Thermotoga. These results suggested that the thermostable nature of XI of A. kamchatkensis G10 may be advantageous in industrial applications and food processing.

• 대한약리학회지
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• 제32권2호
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• pp.269-281
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• 1996

Pharmacokinetics of prednisolone and prednisone undergoing reversible interconversion were analyzed from the model including this metabolic process. Blood samples were drawn serially upto 12 hours after I,V. bolus injection of 1 mg/kg prednisolone sodium phosphate and prednisone into 8 dogs as a crossover manner. Plasma concentrations of those two steroids were simultaneously measured with the method of HPLC. After injection, plasma concentrations of administered prednisolone and prednisone were declined with a biexponential pattern and their metabolic partner was rapidly formed. Plasma concentrations of those metaboite were decayed in parallel with their parent steroids throught the elimination phase. Apparent clearances of prednisolone and prednisone were $11.1{\pm}2.0\;ml/min/kg$ and $45.9{\pm}6.4\;ml/min/kg$, and they were underestimated by 29.4% and 33.6% compared to their real clearances$(15.7{\pm}4.4\;and\;69.2{\pm}17.7\;ml/min/kg)$ estimated using reversible interconversion model. Apparent volume of distribution of prednisolone$(1.32{\pm}0.43\;L/kg)$ and prednisone$(4.81{\pm}2.75\;L/kg)$ were overestimated by 53.5 and 52.7% and were compared to the real volumes $(0.86{\pm}0.30\;and\;3.15{\pm}2.13\;L/kg)$. Mean residence time of prednisolone$(2.0{\pm}0.61\;h)$ and prednisone$(1.74{\pm}0.74\;h)$ were much longer than the real sojourn time$(0.93{\pm}0.26\;and\;0.88{\pm}0.54\;h)$. Essential clearances In the reversible interconversion were greater as following orders: $Cl_{21}$(44.3 ml/min/kg) > $Cl_{20}$(24.2 ml/min/kg) > $Cl_{12}$ (7.9 ml/min/kg) > $Cl_{10}$(7.8 ml/min/kg). Estimated mean values of RF, EE, $%X^1_{ss}$ and $RHO^2_1$ were $0.31{\pm}0.10$, $1.49{\pm}0.23$, $69.3{\pm}16.7%$ and $0.65{\pm}0.10$, respectively. These results suggested that true pharmacokinetic parameters estimated from the model including reversible interconversion were significantly different from the apparent parameters estimated from the conventional mamillary model, and disposition of these two steroids seemed to be well explained by the model including reversible interconversion.

• 한국미생물·생명공학회지
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• 제30권4호
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• pp.359-366
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• 2002

B. polyfermenticus SCD와 상업적으로 사용되는 생균제들의 pH 2.0, 3.0에서 내산성을 비교한 결과, 2시간에서의 생존율은 pH 2.0, 3.0에서 각각 85.6과 92.6%의 생존율을 나타내었으며 4시간에서도 각각 62.8%와 81.2%의 높은 생존율을 나타내었다. B. polyfermenticus SCD의 인공담즙산에 대한 내성을 확인한 결과, 인공담즙산이 함유되지 않은 대조구($3.2$\times$10^{7}$ CFU/ml)보다 균수($6.5$\times$10^{7$ CFU/ml)가 더 많이 증식되었다. 또한 B. polyfermenticus SCD는 비타민 $B_1$, $B_2$를 생산하고 장내유해균웨 의한 비타민 B군의 분해를 억제하여 영양을 공급한다. 게다가 비타민 C를 다량 생산하여 공급하므로 비타민 C의 항산화작용에 의한 피부노화 방지 및 항암작용 등의 여러 가지 효능들도 기대할 수 있다. B. polyfermenticus SCD의 소화력을 다른 Bacillus sp.과 비교했을 때 지방소화력은 비슷한 활성을 나타내었으나, 전분 소화력은 약 2배, 단백소화력은 약 25배의 높은 소화력을 가지고 있었다. B. polyfermenticus SCD는 낮은 pH에서도 안정성이 확보되었으며 저장온도는 가혹조건($40^{\circ}C$, RH 70%)에서 26.8%거 비교적 낮은 안정성을 나타내었을 뿐 저온 및 실온에서는 매우 안정하였다. 그리고 수중에서와 5% glucose용액, 고농도의 NaCl 에서도 매우 높은 안정성을 나타내어 식품을 포함한 다양한 형태의 제품에 적용했을 때에도 그 안정성이 탁월하여 응용성이 크다고 할 수 있겠다. B. polyfermenticus SCD복용에 따른 장내균총의 변화에서는 Salmonella, E. coli, Staphylococcus등 유해균의 유의성 있는 감소효과를 나타내었으며, 유익균과의 공생작용과 유익균에 대한 생육촉진작용을 나타내었다. 장내 암모니아의 변화에서는 복용 중에 유의성 있는 감소효과를 보였다. 이상의 결과로서 B. polyfermenticus SCD의 생균제로서의 특성은 기능성 식품의 개발 등 산업적으로 매우 유용하다고 판단된다.

• 한국미생물·생명공학회지
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• 제36권2호
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• pp.106-114
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• 2008

김치로부터 강력한 H. pylori 생육 저해활성을 보이는 균주를 분리, 동정하여 Lb. plantarum NO1으로 명명하였다. Lb. plantarum NO1은 H. pylori 뿐만 아니라 그람 양성균 및 그람 음성균들에 넓은 범위의 저해활성을 나타내었다. Lb. plantarum NO1의 배양 상징액을 H. pylori배양액에 첨가한 후 H. pylori의 urease 활성을 측정한 결과 Lb. plantarum NO1의 강한 urease 억제활성($40{\sim}60%$ 저하)을 확인할 수 있었다. AGS위암 세포주에 H. pylori를 부착시킨 후 Lb. plantarum NO1의 배양액을 첨가하여 AGS세포에서 H. pylori 탈착능을 측정한 결과 Lb. plantarum NO1은 유산균 배양액 무첨가보다 33% 이상 높은 H. pylori 탈착능을 나타내었으며, 비교구로 사용된 Lb. rhamnosus GG, Lb. sakei SI3에 비해 더 우수한 H. pylori 탈착능을 나타내었다. 분리균주의 장내 생존성 여부 확인을 위하여 내산성, 인공위액에서 2시간동안 처리한 결과 Lb. plantarum NO1이 초기균수$(10^9CFU/ml)$를 유지하면서 높은 저항성을 나타내었다. Oxgall 농도 0.3%와 0.5%의 인공담즙에서 24시간 처리한 후에도 초기균수$(10^9CFU/ml)$를 유지하였다. 뿐만 아니라 인공위액에서 생존한 균주를 연속적으로 인공담즙으로 처리하였을 때에도 높은 생존율$(10^8{\sim}10^9CFU/ml)$를 유지하였다. Lb. plantarum NO1의 용혈성 반응 유무 결과 용혈반응이 일어나지 않았으므로 인체에 안전하다는 것을 간접적으로 확인할 수 있었다. 본 연구에서 김치로부터 분리한 H. pylori억제 유산균 Lb. plantarum NO1은 장내에 생존 가능성도 높으며, 동시에 위에서 효과적으로 H. pylori를 억제 할 수 있을 것으로 기대되어진다.

• 대한화학회지
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• 제4권1호
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• pp.18-26
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• 1957

There are two problems to be solved by our efforts in the enamel frit. One is how we can cover the enamel frit thin with complete milk white as possible, and the other is how it can be, made resistant for chemicals than before one. The frit which can solved the two problems just mentioned above is titanium enamel frit. This frit has been developed in America after War Ⅱ, and now the research for concerning antimony frit into titanium frit is under development entirely. In order to develope the enamel industry in Korea, it is urgent problem to convert antimony frit into titanium frit. By the way the titanium frit is emulsified titanium oxide crystal which made through reheating the supersaturated solution of titanium oxide in the basis of glass. Unfortunately, there are many obscure points in active fact or which influence on its composition and characteristics yet. However, this task was tried for the first in Korea. As first step, the test was carried on the reference books, and we can be possible convert antimony frit into titanium frit as a result of this experiment. As a conclusion, for the purpose of developing the enamel industry in Korea, we studied that the research for converting antimony enamel frit which has been used popularly into titanium enamel frit which is more economic and resistant for chemicals. As a result of experiments, the following points concerning with titanium frit have become clearly. 1. It is better when the composition of titanium enamel frit has as following table.Man Duck San Silica 24 An Yang Feldspar 20 Borax 28 Sodium Nitrate 4 Cryolite 7 Calcium Carbonate 3.6∼1 Titanium Oxide 10 Calcium phosphate 0 ∼3.2 Calcium Fluoride 0∼1.8 Antimony Oxide 0∼0.5 2. The amount of $TiO_2$, to be added is $10%\;to\;12{%,\;CaF_2\;is\;under\;1.8%,\;P_2O_5\;is\;under\;1.6%,\;Sb_2O_3\;is\;under\;0.5%$. 3. In the titanium frit, the limit of iron oxide amount to be included is under 0. 5%. 4. Comparing the titanium enamel frit with antimony enamel frit not only the titanium frit can be savely 20.6% in the price of raw materials, but one time of glazing and heating process is omitted in each case, and it is known the titanium frit is more resistant for chemicals than antimony frit.