• Applied Biological Chemistry
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• 제38권5호
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• pp.463-467
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• 1995

생쥐(Balb/C) 간과 신장의 microsomal cytochrome P-450 효소계에 의한 ${\alpha}$-endosulfan의 시험관내 대사시험을 수행하였다. ${\alpha}$-Endosulfan은 endosulfan lactone(EL), endosulfan hydroxyether(EHE), endosulfan alcohol(EA), endosulfan sulfate(ES), endosulfan ether(EE) 그리고 ${\beta}$-endosulfan(${\beta}$-E) 등으로 대사되었으며 주요 대사산물은 간에서 EL(13.2%) 및 EA(11.5%)이었으며 신장에서 EA(17.4%) 및 EHE(19.3%)이었다. Microsome 배양액중 유기용매 추출성 대사산물은 63.4%이었으며 수용성 대사산물은 37.1%이었다. 수용성 대사산물은 EA(83.9%), EHE(4.5%) 그리고 ES(2.3%)로서 주요 수용성 대사산물은 EA이었다. Piperonyl butoxide는 ${\alpha}$-endosulfan으로부터 EE의 생성을 86%, EA의 생성을 92% 그리고 EHE, EL 및 ES의 생성을 대부분 저해하였다.

• Biomolecules & Therapeutics
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• 제32권4호
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• pp.474-480
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• 2024

Streptomyces avermitilis genome includes 33 genes encoding monooxygenation-catalyzing cytochrome P450 enzymes. We investigated the structure of CYP107P2 and its interactions with terpenoid compounds. The recombinant CYP107P2 protein was expressed in Escherichia coli and the purified enzyme exhibited a typical P450 spectrum upon CO-binding in its reduced state. Type-I substrate-binding spectral titrations were observed with various terpenoid compounds, including α-pinene, β-pinene, α-terpinyl acetate, and (+)-3-carene. The calculated binding affinities (Kd) ranged from 15.9 to 50.8 µM. The X-ray crystal structure of CYP107P2 was determined at 1.99 Å resolution, with a well-conserved overall P450 folding conformation. The terpenoid compound docking models illustrated that the structural interaction between monoterpenes and CYP107P2, with the distance between heme and terpenes ranging from 3.4 to 5.4 Å, indicates potential substrate binding for P450 enzyme. This study suggests that CYP107P2 is a Streptomyces P450 enzyme capable of catalyzing terpenes as substrates, signifying noteworthy advancements in comprehending a novel P450 enzyme's involvement in terpene reactions.

• Journal of Nutrition and Health
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• 제25권1호
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• pp.3-11
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• 1992

This paper examines the effects of dietary fats on the fatty acid composition and market enzyme activites during liver damage in 2-acetylaminofluorene treated rats. Weaning Sprague-Dawley male rats were fed the diet of beef tallow(BT source of sturated fatty acid) corn oil(CO source of n-6 fatty acid) and perilla oil(PO source of n-3 fatty acid) at the level of 15% fat. Ten days after feeding 2-acetylaminofluorene(2-AAF) was injected intraperitoneally twice every week at the level of 50mg/kg body weight for 7 weeks. Liver microsomal and cytosolic fractions were collected to determine the microsomal fatty acid composition lipid peroxide(malondialdehyde MDA) contents glucose 6-phosphatase(G6 Pase) activity cytochrome(Cyt) P-450 contents and cytosolic glutathione S-transferase(G6 Pase) activity cytochrome(Cyt) P-450 contents and cytosolic glutathione S-transferase(GST) activity. The fatty acid composition in microsomal fraction was reflected by different dietary fats. By 2-AAF treatment linoleic acids were increased regardless of the diet MDA contents were higher in CO group than it was in BT group. However 2-AAF treatment decreased MDA contents in all dietary groups. G6Pase activity of BT group was higher than those of the other gropus. CO group had the highest Cyt P-450 contents and 2-AAF treatment lowered Cyt P-450 contents only in CO gropu GST activites were higher in CO than in BT group whereas the enzyme activites were increased by 20AAF treatment in all dietary groups. These results suggest that dietary fats and 2-AAF treatment in all dietary groups,. These results suggest that dietary fats and 2-AAF treatment affect microsomal fatty acid composition The enzyme activities concerned with liver damage were influenced differently by dietary fats and 2-AFF treatment Although PO diet contains much more polyunsaturated fatty acids than CO diet PO diet doesn't cause more oxidant stress compared with CO diet in these data.

• Journal of Microbiology and Biotechnology
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• 제22권1호
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• pp.141-146
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• 2012

Malassezia globosa is a common pathogenic fungus that causes skin diseases including dandruff and seborrheic dermatitis in humans. Analysis of its genome identified a gene (MGL_1677) coding for a putative NADPH-P450 reductase (NPR) to support the fungal cytochrome P450 enzymes. The heterologously expressed recombinant M. globosa NPR protein was purified, and its functional features were characterized. The purified protein generated a single band on SDS-PAGE at 80.74 kDa and had an absorption maximum at 452 nm, indicating its possible function as an oxidized flavin cofactor. It evidenced NADPH-dependent reducing activity for cytochrome c or nitroblue tetrazolium. Human P450 1A2 and 2A6 were able to successfully catalyze the O-deethylation of 7-ethoxyresorufin and the 7-hydroxylation of coumarin, respectively, with the support of the purified NPR. These results demonstrate that purified NPR is an orthologous reductase protein that supports cytochrome P450 enzymes in M. globosa.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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• pp.188.2-189
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• 2003

To develop cytochrome P450 (P450) modulators with low toxicity from natural products, we have evaluated more than 20 compounds in mouse and rat models. Among those, the effects of 1,8-cineole and diallyl sulfide (DAS) were most profound in modulating P450 expression. In this presentation, the effects of 1,8-cineole and diallyl sulfide (DAS) on P450 expression were introduced. (omitted)

• Toxicological Research
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• 제9권1호
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• pp.35-44
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• 1993

Role of rat cytochrome P-450 2E1(P-450 2E 1) in the metabolism of chlorzoxazone was examined by using several approaches' (1) selective inhibiton of catalytic activity in rat liver microsomes by diethyldithiocarbamate, (2) correlation of dimethylnitrosomine N-demethylation with chlorzoxazone 6-hydroxylation, and (3) immunoinhibition of catalytic activity with rabbit anti-rat P-450. The results indicated that P-450 2E1 is responsible for the metabolism of chlorzoxazone.

• Biomolecules & Therapeutics
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• 제27권2호
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• pp.127-133
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• 2019

The study of the genus Streptomyces is of particular interest because it produces a wide array of clinically important bioactive molecules. The genomic sequencing of many Streptomyces species has revealed unusually large numbers of cytochrome P450 genes, which are involved in the biosynthesis of secondary metabolites. Many macrolide biosynthetic pathways are catalyzed by a series of enzymes in gene clusters including polyketide and non-ribosomal peptide synthesis. In general, Streptomyces P450 enzymes accelerate the final, post-polyketide synthesis steps to enhance the structural architecture of macrolide chemistry. In this review, we discuss the major Streptomyces P450 enzymes research focused on the biosynthetic processing of macrolide therapeutic agents, with an emphasis on their biochemical mechanisms and structural insights.

• 대한수의학회지
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• 제38권1호
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• pp.17-28
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• 1998

Drug-metabolizing activities of Korean native cattle and swine were investigated from viewpoints of the cytochrome P-450's level, their dependent mixed function oxidase activities, the reactive oxygen species formation and cytosolic enzyme acitivities from each liver homogenates. Level of cytochrome P-450 in the liver microsome of Korean native cattle was $0.28{\pm}0.05nmole/mg$ and that in pigs $0.35{\pm}0.03nmole/mg$. Level of cytochrome $b_5$ of Korean native cattle was $0.24{\pm}0.06nmole/mg$, and that of pigs $0.2{\pm}0.05nmole/mg$, showing no difference between two species. NADPH P-450 reductase were higher in Korean native cattle ($58.3{\pm}5.3nmole/mg/min$) than in pigs ($29.9{\pm}3.8nmole/mg/min$)(p<0.01). The activities of cytochrome P-450 dependent monooxygenases such as ethoxyresorufin O-deethylase (cattle, $96.5{\pm}12.5nmole/mg/min$ ; pigs, $13.6{\pm}2.1nmole/mg/min$), N-benzphetamine N-demethylase (cattle, $5.23{\pm}0.82nmole/mg/min$ ; pigs, $0.76{\pm}0.3nmole/mg/min$) and aniline hydroxylase (cattle, $0.95{\pm}0.1nmole/mg/min$ ; pigs, $0.33{\pm}0.08nmole/mg/min$) were much higher in Korean native cattle than in swine(p<0.01). However, the activity of testosterone $7{\alpha}$-hydroxylase was higher in swine ($90.4{\pm}1.2nmole/mg/min$) than cattle (cattle, $32.8{\pm}1.2nmole/mg/min$). Interestingly, testosterone $16{\alpha}$-hydroxylase, a marker enzyme for P-450 IIA was not detected in both animal species. These results suggest that Korean native cattle and pigs have high contents of P-450 IA1 and P-450 IIIA. Total sulfhydryl compound (cattle, $10.3{\pm}1.1nmole/mg$ ; Pigs, $14.5{\pm}1.8nmole/mg$) and glutathione related enzymes except glutathione reductase (cattle, $38.1{\pm}7.9nmole/mg/min$; swine, $22{\pm}3.6nmole/mg/min$) showed higher levels in swine than in Korean native cattle. Superoxide dismutase (cattle, $7.64{\pm}0.84nmole/mg/min$ ; pigs, $4.47{\pm}0.94nmole/mg/min$) and catalase (cattle, $30.4{\pm}3.7nmole/mg/min$ ; pigs, $17.2{\pm}1.8nmole/mg/min$) were remarkably higher in Korean native cattle than in swine (p<0.05).

• 사상체질의학회지
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• 제24권4호
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• pp.84-91
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• 2012

Objectives : The purpose of this study is to investigate the inhibitory or inductive potentials of Yuldahanso-tang (YDT) and Chungsimyonja-tang (CST), herbal formulas for Taeeumin, on cytochrome P450 (CYP450) drug metabolizing enzyme. The mechanisms for the herbal formula-drug interaction has not been well reported in spite of the chance for co-administration with conventional drugs. Methods : To evaluate the interaction potential of YDT-drug or CST-drug, the fluorescence-based enzyme assays on CYP450 isozymes including CYP3A4, CYP2C19, CYP2D6 and CYP2E1 were established in vitro. The inhibitory effects of herbal formulas were characterized with $IC_{50}$ values. Results : YDT showed inhibitory effects on CYP2D6 and CYP2E1-mediated metabolism, while it exhibited week inhibition on CYP3A4 and CYP2C19 relatively. CST exerted relatively weak inhibitory effects on the four CYP450 isozymes compared to that of YDT. Conclusions : These results suggest that the herbal formula-drug interaction could be occur when YDT are co-administered with drugs mediated by CYP2D6 or CYP2E1.

• Journal of Microbiology and Biotechnology
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• 제27권5호
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• pp.983-989
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• 2017

NADPH-P450 reductase (NPR) transfers electrons from NADPH to cytochrome P450 and heme oxygenase enzymes to support their catalytic activities. This protein is localized within the endoplasmic reticulum membrane and utilizes FMN, FAD, and NADPH as cofactors. Although NPR is essential toward enabling the biochemical and pharmacological analyses of P450 enzymes, its production as a recombinant purified protein requires a series of tedious efforts and a high cost due to the use of $NADP^+$ in the affinity chromatography process. In the present study, the rat NPR clone containing a $6{\times}$ Histidine-tag (NPR-His) was constructed and heterologously expressed. The NPR-His protein was purified using $Ni^{2+}$-affinity chromatography, and its functional features were characterized. A single band at 78 kDa was observed from SDS-PAGE and the purified protein displayed a maximum absorbance at 455 nm, indicating the presence of an oxidized flavin cofactor. Cytochrome c and nitroblue tetrazolium were reduced by purified NPR-His in an NADPH-dependent manner. The purified NPR-His successfully supported the catalytic activities of human P450 1A2 and 2A6 and fungal CYP52A21, yielding results similar to those obtained using conventional purified rat reductase. This study will facilitate the use of recombinant NPR-His protein in the various fields of P450 research.