• Natural Product Sciences
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• 제9권4호
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• pp.235-240
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• 2003

Tectorigenin has an apoptosis-inducing ability and immunosuppressive activity. We investigated the effect of tectorigenin on Phase I and II enzyme activities to elucidate the pharmacological action of the immunosuppressive tectorigenin in the diabetic rat. This compound was obtained from the hydrolysis of tetoridin isolated from the flower of Pueraria thumbergiana (Leguminosae). This crude drug (Puerariae Flos) has been used as a therapeutic for diabetes mellitus in traditional Korean medicine. Tecotrigenin inhibited the formation of malondialdehyde (MDA) and hydroxy radicals in serum and liver but promoted superoxide dismutase (SOD) activity. Low MDA contents and low xanthine oxidase and aldehyde oxidase activities were observed in the tectorigenin-treated rats, suggesting that such Phase I enzyme activities are the major source of lipid peroxidation. However, tectorigenin increased Phase II enzyme activities such as SOD, glutathione peroxidase and catalase, suggesting the activation of free radical-scavenging enzymes. The activities of tectorigenin were comparable to those of glibenclamide, which was employed as a positive control. These results suggest that tectorigenin may share some biological properties with glibenclamide in insulin-dependent-diabetes mellitus (IDDM).

• Asian-Australasian Journal of Animal Sciences
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• 제14권2호
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• pp.231-236
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• 2001

This study was conducted to investigate the effect of supplemented enzyme complex on growth performance and nutrient digestibility in pigs weaned at 14 days of age. Eighty pigs ($4.02{\pm}0.11kg$ of average body weight) were allotted in a completely randomized block design. Treatments were as follows: 1) control (negative), 2) control (positive, $Kemzyme^{(R)}$), 3) 0.1%, 4) 0.2% and 5) 0.3% of newly developed enzyme complex. Each treatment has 4 replicates with 4 pigs per replicate. During phase I period (d 0 to 14), ADG and ADFI were numerically higher in pigs fed diets supplemented enzyme complex regardless of their inclusion levels compared to pigs fed control (negative) diet. Feed/gain (F/G) was also better in pigs fed enzyme complex diet than that of pigs fed control (negative) diet. In addition, with increasing the inclusion level of enzyme complex, ADG and ADFI were improved. However, there was no significant difference between treatment in all growth parameters. During phase II period (d 15 to 28), ADG, ADFI and F/G showed the same tendency as in phase I period. For overall period (d 0 to 28) ADG was highest in pigs fed diet included 0.2% enzyme complex in all treatments but not significantly different. During phase I period, the digestibilities of all nutrients did not showed any significant difference between treatments. However, pigs fed diet contained enzyme complex and positive control diet (Kemzyme) showed numerically higher nutrient digestibilities in all nutrients than pigs fed negative control diet. During phase II period, data were consistent with those observed in phase I period. Especially, the digestibility of phosphorus was significantly higher in pigs fed diets contained enzyme complex including phytase than pigs fed control (negative and positive) diets (p<0.05). For overall experimental period, fecal or ileal amino acid digestibility were not affected by dietary treatment. Enzyme complex newly developed and used in this study can be possibly recommended as a growth promoter when supplemented in diet for early weaned piglets.

• 대한수의학회지
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• 제28권2호
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• pp.321-325
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• 1988

This experiment was carried out to determine the progesterone concentration of bovine plasma by liquid phase double antibody enzyme immunoassay. The optimum conditions of assay-system, double (first and second) antibody and carrier (normal rabbit serum) were investigated. The optimum dilution rate of first antibody, second antibody and normal rabbit serum was $10{\times}10^3$ to $15{\times}10^3$, 20 and $1{\times}10^3$ times, respectively.

• BMB Reports
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• 제30권1호
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• pp.60-65
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• 1997

Expression of human Cu.Zn-superoxide dismutase (SOD) with activity comparable to human erythrocyte enzyme was achieved in E. coli B21(DE3) by using the pET-17b expression vector containing a T7 promoter. Recombinant human SOD was found in the cytosol of disrupted bacterial cells and represented > 25% of the total bacterial proteins. The protein produced by the E. coli cells was purified using a combination of ammonium sulfate precipitation, Sephacryl S-100 gel filtration and DEAE-Sephacel ion exchange chromatography. The recombinant Cu,Zn-SOD and human erythrocyte enzyme were compared using dismutation activity, SDS-PAGE and immunoblotting analysis. The mass of the subunits was determined to be 15,809 by using a electrospray mass spectrometer. The copper specific chelator. diethyldithiocarbamate (DOC) reacted with the recombinant Cu,Zn-SOD. At $50{\mu}M$ and $100{\mu}M$ concentrations of DOC, the dismutation activity was not inhibited for one hour but gradually reduced after one hour. This result suggests that the reaction of DOC with the enzyme occurred in two distinct phases (phase I and phase II). During phase I of this reaction, one DOC reacted with the copper center, with retention of the dismutation activity while the second DOC displaced the copper, with a loss of activity in phase II.

• Biomolecules & Therapeutics
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• 제14권4호
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• pp.226-234
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• 2006

Disturbance of antioxidant system is very common in chronic alcoholics and herbal or natural products with antioxidant activity have been used for its treatment. This study was to investigate the effect of Vitis vinifera extract(V), Schisandra chinensis extract(S), Taraxacum officinale extract(T), Gardenia jasminoides extract(G), Angelica acutiloba extract(A) and Paeonia japonica extract(P), and their combinations on the antioxidant and ethanol oxidation system. Male Sprague-Dawley rats were subjected to Lieber-DeCarli ethanol liquid diet(ED) and were then given different herbal extract mixtures for 6 weeks including VST(V 100+S 150+T 150mg/kg/day), VSG(V 100+S 150+G 150mg/kg/day), VTG(V 100+T 150+G 150mg/kg/day), and VAP(V 100+A 150+P 150mg/kg/day). When the activity of alcohol dehydrogenase(ADH) and acetaldehyde dehydrogenase(ALDH) were compared between ED only group and herbal extracts treatment group, the differences were statistically significant. Phase I and II(glutathione-S-transferase, phenol sulfatransferase) enzyme activities were found to be significantly higher in the VAT treatment group compared to the ED group. Herbal extracts not only repressed the ethanol-induced elevation of malondialdehyde level, but also protected against ethanol-induced decrease in glutathione content, glutathione reductase, glutathione peroxidase, catalase and superoxide dismutase activities. The administration of the herbal extracts was found to be effective in eliminating lipid-peroxides induced by long-term consumption of alcohol by activating various enzyme systems and physiological active compound formation system. After a chronic consumption of alcohol, Angelica Radix protected the liver via activating the ethanol-metabolism enzyme system, and Paeoniae Radix via activating the ethanol-metabolism enzyme and the phase I, II-metabolism enzyme system. Taraxaci Herba was also effective in liver protection via activating the ethanol-metabolism enzyme system and the phase I, II-metabolism enzyme system, Gardeniae Fructus via activating the phase II-metabolism enzyme system and the anti-oxidation system enzyme, and Schisandra Fructus and a grapestone via activating the anti-oxidation system. Our data suggest that these herbal extracts may be useful as a health functional food or new drug candidate for fatty liver and hepatotoxicity induced by chronic alcohol consumption.

• Korean Journal of Acupuncture
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• 제18권1호
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• pp.11-20
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• 2001

댑싸리하고초 약침액을 이용하여 phase II detoxification 효소인 quinone reductase (QR 및 GST) 유도, GSH 생성량, phase I 효소 cytochrome P4501A1 활성억제, 발암물질 B[a]P-DNA adduct 형성의 저해효과 등을 측정하였다. QR 생성 유도를 Hepa1c1c7로 실험한 결과, 댑싸리하고초 약침액및 열수추출액 모두에서 유도 되었으며, 농도가 높아질수록 유도율이 더 높게 나타났으며, GSH와 GST의 유도율도 약침액에서 나타났다. 댑싸리하고초 약침액 $5{\times}$에서 45.2%의 cytochrome P4501A1 효소활성 저해효과를 측정할 수 있었다. 또한 B[a]P-DNA binding 저해효과를 실험한 결과, 약침액 $3{\times}$ 농도에서 45.0%의 저해효과가 있었다. 이상의 결과에 의하면 댑싸리하고초 약침액은 암억제 물질로서의 기능을 충분히 발휘할 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제22권3호
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• pp.339-342
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• 2012

This study describes the characterization of a new angiotensin I-converting enzyme (ACE) inhibitory peptide from a Korean traditional rice wine. After purification of the ACE inhibitor peptides with ultrafiltration, Sephadex G-25 column chromatography, and successively $C_{18}$ and SCX solid-phase extraction, reverse-phase HPLC, and size exculsion chromatography, two types of the purified ACE inhibitors with $IC_{50}$ values of 0.34 mg/ml and 1.23 mg/ml were finally obtained. The two purified ACE inhibitors (F-1 and F-2) were found to have two kinds of novel oligopeptides, showing very little similarity to other ACE inhibitory peptide sequences. The amino acid sequences of the two purified oligopeptides were found to be Gln-Phe-Tyr-Ala-Val (F-1) and Ala-Gly-Pro-Val-Leu-Leu (F-2), and their molecular masses were estimated to be 468.7 Da (F-1) and 357.7 Da (F-2), respectively. They all showed a clear antihypertensive effect on spontaneously hypertensive rats at a dosage of 500 mg/kg.

• 한국환경보건학회지
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• 제27권2호
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• pp.10-16
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• 2001

To evaluate an effect of circadian variation on the xylene metabolizing enzyme activities, 50% m-xylene in olive oil(0.25 $m\ell$/100g body weight) was intraperitoneally administered to the rats every other day for 6 days both in the night; 24:00 and the day; 12:00. Then animals were sacrigiced at 8hr after last injection of m-xylene. Hepatic microsomal cytochrome p450 contents were more increased both in control and xylene treated rats of night phase than those of day phase. But the activity of hepatic alcohol dehydrogenase(ADH) in control of night phase showed the similar value with that in those of day phase and xylene treated rats of day phase showed an increasing tendency of hepatic ADH activity as those of night phase showing similar activity. Furthermore, control rats of night phase than those of day phase. And by xylene treatment, enzyme activities of rats of day phase were higher tendency in rats of control but those of night phase were somewhat inhibited. Besides, xylene-treated animals of night phase showed increasing tendency of urinary methylhippuric acid concentration compared with those of day phase. On the other hand, liver weight per body weight(%), hepatic lipid peroxide content and serum xanthine oxidase activity were higher in night phase. And the activities of hepatic oxygen free radical metabolizing enzymes such as xanthine oxidase, gluthathione S-transferase, and xylene-treated rats of night phase than those of day phase. In conclusion, it can be hypothesized on the basis of the results that the accumulation rate of m-xylene intermediate metabolite, i.e. m-methylbenzaldehyde in liver tissus may be higher in night phase than in day phase and it may be responsible for higher liver toxicity in bight phase than in day phase.

• Journal of Microbiology and Biotechnology
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• 제2권3호
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• pp.189-196
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• 1992

A $\alpha$-l, 4-D-glucan maltohydrolase $(\beta$-amylase), secreted by the mesophilic aerobic bacterium Bacillus polymyxa No.26, was purified and characterized. The enzyme production was increased after a logarithmic phase of bacterial growth and paralleled with the onset of bacterial sporulation. By applying anion exchange chromatography and gel filtration the enzyme was purified 16.7-fold and had a specific activity of 285.7 units/mg. Two enzyme activities were eluted on a column of DEAE-Sephadex chromatography, and they were designated as E-I for a major enzyme peak and E-II for a minor peak. Of them, E-I enzyme peak was further purified by using gel chromatography. The molecular mass of this enzyme was determined to be 64, 000 daltons and consisted of a single subunit, showing an isoelectric point of 8.9. The enzyme was able to attack specifically the $\alpha$-l, 4-glycosidic linkages in soluble starch and caused its complete hydrolysis to maltose and $\beta$-limited dextrin. This amylolytic enzyme displayed a temperature optimum at $45^\circ{C}$ and a pH optimum at 7.0. The amino acid composition of the purified enzyme was quite similar to the other bacterial $\beta$-amylases reported. Surprisingly, the purified enzyme from this aerobe only exhibited hydrolytic activity on soluble starch, not on starch granules. The degradation of from starch by $\beta$-amylase was greatly stimulated by pullulanase addition. These results differentiated from other $\beta$-amylases reported. Based on a previous result that showed the enzyme system involves in effective degradation of raw starch granules, this result strongly suggested that the purified enzyme (E-I) can be a synergistic part of starch granule-digestion and E-II plays a crucial role in digestion of starch granules.

• 한국미생물·생명공학회지
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• 제31권1호
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• pp.51-56
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• 2003

안정성이 확보된 식품에서 ACE저해 활성 물질을 검색하는 연구의 일환으로, 옥수수 글루텐을 Flavourzyme, Pescalase, 그리고 Thermolysine/Pescalase 등으로 가수분해하여 얻은 가수 분해물로부터 ACE 활성 저해 펩타이드를 다음과 같은 과정으로 분리, 정제하였다. 10% 에탄올로 평형화된 ODS chromatography를 이용 단백질 분획들을 얻고, Bio-Gel P-2 column과 reverse phase HPLC를 통해 5개의 ACE 저해 펩타이드를 분리, 정제하였다. 그 아미노산 서열은 LPF($IC_{50}$ = 40 $\mu$M), GPP($IC_{50}$ = 17.6 $\mu$M), PNPY($IC_{50}$ = 30.7 $\mu$M), SPPPFYL($IC_{50}$/ = 63 $\mu$M), and SQPP($IC_{50}$ = 17.2 $\mu$M)로 밝혀졌다. 이 펩타이드들은 경구투석 시 가수분해 효소에 대응하여 체내에서 안정성이 뛰어나고, 소장에서도 쉽게 흡수될 것으로 사료되어 상시 섭취하는 식품이나 음료에 첨가하여 이용한다면 그 유용성이 기대된다.