• Proceedings of the Korean Society of Applied Pharmacology
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• 1992.05a
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• pp.23-23
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• 1992

횐 쥐에서 sulfotransferase 양성균주를 검색한 결과 호기성 조건보다는 혐기성 조건의 균이 많은 것으로 나타났다. 이러한 곁과는 장내미생물의 대부분이 혐기성균이라는 것과도 일치하는 것이다. 검색용 배지에서 형광을 나타내는 Sulfotransferase를 생산하는 4개 균주에서 가장 활성이 높은 k-36균주에 대하여 동정하였다. K-36 균주는 혐기적 및 호기적 양조건에서 잘 자라는 통성혐기성균이고, Gas를 생산하며 포자를 생성하지 않는 Gram음성의 간균이었다. 이것으로 보아 이균은 Enterobacteriacease의 Klebsiella oxytoca 로 동정했다. K-36에서 분리한 Sulfotransferase의 반응양식은 p-nitrophenylsulfate와 p-nitrophenol이 반응시간과 함께 동량씩 감소하여 반응 생성물인 phenylsulfate와 p-nitrophenol을 생성시켰으며 sulfatase 반응은 진행되지 않았다.

• Archives of Pharmacal Research
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• v.13 no.1
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• pp.51-54
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• 1990

Effect of scoparone (6, 7-dimethoxycoumarin) on the hepatic cytosolic sulfotransferase activity was investigated. After treatment with scoparone, hepatic cytosolic sulfotransferase activity was increased with odse and time-dependent manner as compared to control. The $V_{max}$ value (control = 1.33 n moles/mg protein/min, scoparone = 2.39n moles/mg protein/min) without affecting the $K_m$ value for p-nitrophenol was increased by the scoparone treatment. Whereas, the hepatic cytosolic sulfotransferase was not changed by the addition of scoparone in vitro, and was strongly inhibited by the addition of metabolites of scoparone. The results obtained suggest that the characteristics of increase in the enzyme activity may include induction of enzyme proteins, and may be due to the metaboltes of scoparone.

• YAKHAK HOEJI
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• v.36 no.2
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• pp.167-172
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• 1992

Klebsiella K-36 producing novel sulfotransferase was isolated from rat intestinal flora. The novel sulfotransferase catalyzed the transfer of sulfate group from p-nitrophenylsulfate to phenolic compounds but it did not use PAPS(3'-phosphoadenosine 5'-phosphosulfate) as a donor substrate. The present enzyme was 160 K daltons. Optimal pH was 10. When p-nitrophenyl sulfate was used as a donor substrate, 1-naphthol was the best substrate, followed by phenol, phenanthrol and tyrosine. The apparent Km for phenol using p-nitrophenylsulfate as a donor substrate and that for p-nitrophenylsulfate using phenol as an acceptor substrate were determined to be 0.66 mM and 0.11 mM, respectively.

• YAKHAK HOEJI
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• v.36 no.5
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• pp.455-459
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• 1992

Haemophilus K-12 producing novel sulfotransferase was isolated from mouce intestinal flora. The enzyme catalyzed the transfer of sulfate group from p-nitrophenylsulfate to phenolic compounds. The optimum medium condition for the sulfotransferase production was 0.2% sucrose, 1% yeast extract, $Na_{2}HPO_4$ and 0.5% NaCl. The enzyme production was induced by donor substrate, but was not by accepters. When p-nitrophenylsulfate was used as a donor substrate, 1-naphthol was best substrate, followed by phenol, p-acetaminophenol and tyramine.

• YAKHAK HOEJI
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• v.42 no.1
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• pp.53-58
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• 1998

A novel type of sulfotransferase, arylsulfate sulfotransferase (EC 2.8.2.22) purified from Haemophilus K-12, an intestinal bacterium of a mouse, was immobilized onto AH-S epharose 4B, CH-Sepharose 4B and DEAE-celluose. The enzyme was stabilized for storage more markedly by covalent immobilization onto AH-Spharose 4B or CH-Sepharose 4B and by adsorptive immobilization onto DEAE-cellulose than the free enzyme. The optimal pH and acceptor substrate specificity of immobuized enzyme were similar to those of the free enzyme.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.257-257
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• 1994

생쥐의 장내세균으로 부터 황산전이 반응을 촉매하는 효소인 sulfotransferase를 생산하는 균주를 분리하였으며 동정결과 Haemophilus속 균주로 확인되어 Haemophilus K-12라 명명하였다. 균의 성장과 효소활성과의 관계를 보면 균은 10시간에서 완전히 성장하였으며 효소활성도 이와 비례하였다. Haemophilus K-12의 배지조성에 따른 sulfotransferase의 생산성을 Brain Heart Infusion 배지에서의 생산성과 비교해보면 탄소원으로는 sucrose가 0.2%농도에서 584%로 가장 좋았으며 질소원으로는 yeast extract가 266%로 가장 좋았다. 공여체로 PNS를 최종농도 1mM로 하여 배지에 첨가하였을때 212%로 가장 높은 효소증가를 보였다. 2가금속이온에 의한 효소증가는 현저하지 않았으며 $Mn^{2+}$이 107%로 가장 높았고$Zn^{2+}$와 EDTA에 의해서는 저해를 받았다. 이상의 결과를 종합하여 균배양을 위한 이상적인 배지조성을 sucrose 0.2%, yeast extract 1%, $Na_2$HPO$_4$ 0.25%, NaCl 0.5%로 결정하였다. 결정된 최적배지에 균을 10L 배양하여 초음파 처리, 원심분리한 것을 70 % ammonium sulfate fractionation, DEAE-cellulose column chromatography를 2회, Hydroxyapatite column chromatography, chromatdfocusing column chromatography, Silica PAE column chromatography, Sephacryl S-300 superfine column chromatography를 행한결과 specific activity가 6.76 umoie/min/mg protein인 효소액을 얻었으며 homogeneous enzyme였다. 이렇게 해서 얻은 효소를 이용하여 수용체 기질 특이성을 측정한 결과 1-naphthol이 phenol을 100%로 하였을 때 233%로 가장 좋았으며 Eubacterium A-44 sulfotransferase의 좋은 기질이었던 p-acetaminophenol, tyramine, 9-phenanthrol은 좋은 기질이 되지 못했다. 이상의 결과로 미루어 보아Haemophilus K-12 sulfotransferase는 지금까지 보고된 bacterial sulfotrasferase와는 다른 효소로 사려되며 효소반응기전의 규명이 이루어지면 산업적 응용이 가능할것으로 사료된다.

• Biomedical Science Letters
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• v.11 no.4
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• pp.503-508
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• 2005

Possible mechanisms of increased serum aryl sulfotransferase (AST) isozyme activities in cholestatic rats were studied. Serum AST-I, II and -III, IV isozymes activities were determined from the experimental rats with common bile duct ligation (CBDL) or choledocho-caval shunt (CCS). The activities of serum AST-I, II and -III, IV isozymes were found to be increased significantly in both the CCS plus taurocholic acid (TCA) injected group, and the CBDL plus TCA group than those in each control group, such as CCS or CBDL alone groups. The above results suggest that the elevated serum AST most likely due to increased hepatocyte membrane permeability caused by TCA mediated liver cell necrosis.

• Biomedical Science Letters
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• v.11 no.1
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• pp.37-43
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• 2005

The possible mechanisms of increased aryl sulfotransferase (AST) isozymes activities in cholestatic rat liver were studied. Hepatic AST-I, II and -III, IV activities were determined from the experimental rats with common bile duct ligation (CBDL). The Michaelis-Menten constants in these hepatic enzymes were also measured. The activities of mitochondrial AST-I, II and -III, IV, and microsomal AST-III, IV as well as their Vmax values were found to be increased significantly in CBDL plus taurocholic acid (TCA) injected group than in the control group, such as CBDL alone groups. However, their Km values in the experimental groups did not vary. The results suggest that TCA stimulates biosynthesis of the AST in the liver.

• Archives of Pharmacal Research
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• v.21 no.4
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• pp.475-477
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• 1998

This study is to predict the possible roles of the aryisulfate sulfotransferase (ASST) in the microorganism. At first we studied the spectrum of a distribution of the ASST enzyme through about 1,300 bacteria and the several selected strains were compared with Klebsiella K-36 previously reported in the level of DNA homology using the Southern blot method. From this study, we could predict that this enzyme would not exist in specific bacteria and it might not be a critical enzyme for the life of bacteria.

• Archives of Pharmacal Research
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• v.28 no.5
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• pp.561-565
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• 2005

Bacterial arylsulfate sulfotransferase (ASST) catalyzes the transfer of sulfate group from a phenyl sulfate ester to a phenolic acceptor. The promoter region and the transcripti on start sites of Enterobacter amnigenus astA have been determined by primer extension analysis. Northern blot analysis resolved two mRNA species with lengths of 3.3 and 2.0 kb, which correspond to the distances between the transcriptional initiation sites and the two inverted repeat sequences (IRSs). By length, the 3.3 kb RNA could comprise the three-gene (astA with dsbA and dsbB) operon. ASST has three highly conserved cysteine residues. Reducing and non-reducing SDS-PAGE and activity staining showed that disulfide bond is needed for the activity of the enzyme. To identify the cysteine residues responsible for the disulfide bond formation, a series of Cys to Ser mutants has been constructed and the enzymatic activity was measured. Based on the results, we assumed that the first cysteine (Cys349) might be involved in disulfide bond mainly with the second cysteine (Cys445) and result in active conformation.