• Title/Summary/Keyword: Escherichia coli WP2 uvrA

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Expression of Recombinant Human Cytochrome P450 1A2 in Escherichia coli Bacterial Mutagenicity Tester Strain

  • Chun, Young-Jin
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.305-309
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    • 1998
  • Human cytochrome P450 1A2 is one of the major cytochrome P450s in human liver. It is known to be capable of activating a number of carcinogens such as arylamines and heterocyclic amines. In order to develop the new bacterial mutagenicity test system with human P450, a full length of human P450 1A2 cDNA inserted into pCW bacterial expression vector was introduced to Escherichia coli WP2 uvrA strain which is a well-known E. coli strain for bacterial reverse mutagenicity assay. Expressed human P450 1A2 showed typical P450 hemoprotein spectra. Maximum expression was achieved at 48 hrs after incubating at $30^{\circ}C$ in terrific broth containing ampicillin, IPTG and other supplements. High level expression of P450 1A2 in E. coli WP2 uvrA membranes was determined in SDS-PAGE. The well-known mutagens 2-aminoanthracene and MElQ increased the revertant colonies of E. coli WP2 uvrA expressing human P450 1A2 without an exogenous rat hepatic post-mitochondrial supernatant (S9 fraction) in a dose-dependent manner. The results show that the functional expression of human P450 in bacterial mutagenicity tester strain will provide a useful tool for studying the mechanism of the mutagenesis and carcinogenesis of new drugs and environmental chemicals.

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Bacterial Reverse Mutation Assay of Xylooligosaccharide (Xylooligosaccharide의 복귀돌연변이 시험)

  • 오화균;박윤제;이운택;이지완;이창승;류보경;양창근;윤세왕;강부현
    • Journal of Food Hygiene and Safety
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    • v.14 no.3
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    • pp.259-264
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    • 1999
  • To evaluate the bacterial reverse mutation of xylooligosaccharide(XO)s the in vitro Ames test using Salmonella typhimurium (TA9S, TAIOO, TA1535, TA1537) and Escherichia coli (WP2 uvrA) was performed. XO was negative in Ames test with Salmonella typhimurium and Escherichia coli with and without rat liver microsomal enzyme (S-9 fraction). According to the results, XO does not cause bacterial reverse mutation.

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Mutagenic Mechanism of Chloropropanols in Escherichia coli (대장균 변이주를 이용한 Chloropropanol 변이원성 기구의 해석)

  • Song, Geun-Seoup;Han, Sang-Bae;Choi, Dong-Seong
    • Korean Journal of Food Science and Technology
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    • v.31 no.1
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    • pp.246-251
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    • 1999
  • This study was designed to evaluate the mutagenicity and the primary mutagenic mechanism of chloropropanols by using various genotypes of E. coli WP2, E. coli TK and E. coli GW series strains. Chloropropanols showed the low mutagenic activities in E. coli WP2s and WP2 establishing the following order; 2,3-DCP> 3-MCPD>1,3-DCP. As compared with E. coli WP2s, the decrease of mutagenic activity and the increase of survival rate in E. coli WP2 $(WP2s\;uvrA^+)$ suggest that DNA lesions produced by chloropropanols could be easily removed by excision-repair system. From the diminution of mutagenic activity and survival rate in E. coli CM611 (WP2s lexA), it was confirmed that the mutagenesis by chloropropanols was dependent on the SOS-repair system. This fact could be also confirmed from the result that both the mutagenic activity and survival rate in E. coli TK610 (umuC) were much lower than those in E. coli TK603 $(umuC^+)$. In the experiment to examine the possibility that chloropropanols might have effects on the LexA of SOS response negative regulator, there was no variation in ${\beta}-galactosidase$ activities of E. coli GW1105 $[lexA3\;(Ind^-)]$ and GW1107 [lexA51 (Def)] by addition of the compounds, indicating that chloropropanols do not have any effects on the LexA, itself.

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Antimutagenic Mechanism of Water Extract from Rehmannia glutinosa Liboshitz on 4-nitroquinoline 1-oxide Induced Mutagenesis n E. coli B.r (대장균에서 4-nitroquinoline 1-oride의 변이원성에 대한 숙지황 물추출물의 항돌연변이 작용특성)

  • 안병용;한종현;최동성
    • KSBB Journal
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    • v.16 no.5
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    • pp.486-492
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    • 2001
  • The antimutagenic mechanism of the fraction III(RG III)separated from the water extract of Rehmannia glutionosa was investigated by Escherichia. coli GW and B/r strains. RG-III treatment did not affect the ${\beta}$-galactosidase activity E. coli GW-1060, 1106, 1107 and 1105. These results indicated that RG-III did not induce RecA protein amplification and did not also prevent the proteolytic cleavage of LexA. The bio-antimutagenicity and survival effect of RG-III on 4-nitroquinoline 1-oxide(4NQO), N-methyl-N-nitor-N\`-nitrosoguanidine(MNING) were investigate by E. coli B/r strains with have different pathway of DNA repai. RG-III slightly increased the survival of 4NQO-treated WP2, WP2s, WP67, CM561, CM611 cells, but the reactivation of survival cannot ve explained by the repair mode. RG-III caused the decrease of mutagenicity and lethality treated with MNNG in ZA159 despite of the increase in WP2, WP2s, WP67, CW561, CM611. Compared with bio-antimutagenic effects of RG-III on 4NQO, greatly increased antimutagenic effects of RG-III were observed with all the E. coli B/r strains tested, but less active in ZA159. These results suggest that RG-III was identified as a blocking agent for preventing the 4NQO induced mutagenesis, and may act as chl-products.

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Bacterial Reverse Mutation Test of Wild Ginseng Culture Extract (산삼배양추출물의 세균을 이용한 복귀돌연변이시험)

  • Song Si-Whan;Yang Deok Chun;Choung Se Young
    • Journal of Food Hygiene and Safety
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    • v.19 no.4
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    • pp.193-197
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    • 2004
  • To evaluate the bacterial reverse mutation of wild ginseng culture extract, the in vitro Ames test using Salmonella typhimurium (TA100, TA1,535, TA98, TA1,537) and Escherichia coli (WP2 uvrA) were performed with wild ginseng extract at the concentrations 0, 1.6, 8, 40, 200, 1,000, 2,500 and $5,000{\mu}g/ml/plate$. Wild ginseng culture extract was negative in Ames test with both Salmonella typhimurium or Escherichia coli with and without rat liver microsomal enzyme (S-9 fraction). According to these results, we concluded that wild ginseng culture extract did not cause bacterial reverse mutation.

Mutagenicity Studies of the Herbicide Methiozolin (제초제 Methiozolin의 유전독성평가)

  • Koo, Suk-Jin;Lee, Zong-Yun;Park, Cheol-Beom
    • The Korean Journal of Pesticide Science
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    • v.15 no.4
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    • pp.374-382
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    • 2011
  • We investigated the mutagenicity of methiozolin, newly developed herbicide, in vitro reverse mutation test using Salmonella typhimurium and Escherichia coli, chromosome aberration test using chinese hamster lung (CHL) cells and in vivo micronucleus test of mice. In the reverse mutation test, the methiozolin did not induce mutagenicity in Salmonella typhimurium TA98, TA100, TA1535, TA1537, Escherichia coli WP2uvrA with and without metabolic activation at $5,000{\mu}g$/plate. In the chromosome aberration test, the results showed no incidence of increased structural and numerical chromosome abberrations at any doses tested (80, 40, $20{\mu}g$/mL). In micronucleous test, the ratio of micronuclei was measured in polychromatic erythrocytes with treated methiozolin for ICR mice. No incidence of increased micronuclei were observed in polychromatic erythrocytes (1,500, 1,000, 500 mg/kg). Based on these results, we concluded that methiozolin has no mutagenic toxicity in vitro and in vivo systems.

Evaluation of the Mutagenic Properties of Two Lignans from Acanthopanax koreanum Nakai

  • Hong, Chang-Eui;Lyu, Su-Yun
    • Toxicological Research
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    • v.29 no.4
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    • pp.279-283
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    • 2013
  • Acanthopanax koreanum Nakai, a well known traditional herb grown in Jeju Island, South of Korea, has been used as a tonic and sedative agent, as well as in the treatment of diabetes and immune diseases. Mutagenicity of two lignans, syringaresinol and tortoside A isolated from A. koreanum, was assessed using Salmonella/microsome (Ames) test. Tester strains used were Salmonella typhimurium TA98, TA100, TA1535, and Escherichia coli WP2uvrA. The mutagenic activity was determined both in the absence or presence of S9 mixture. As a result, tortoside A did not cause any increase in the number of $his^+$ revertants in S. typhimurium and E. coli WP2uvrA strains in the presence or absence of S9 mix, compared to the controls. Similarly, low concentrations of syringaresinol (750 and 1,500 ${\mu}g$/plate) did not show any mutagenic properties in all bacterial strains, in the presence or absence of S9 mixture. However, in the high concentration of syringaresinol (3,000 ${\mu}g$/plate), the number of revertants were increased in TA1535 strains, in the absence of S9 metabolic activation. Therefore, in vivo experiments such as comet assay are needed to further determine the genotoxic/carciogenic potential of syringaresinol isolated from A. koreanum.

Genotoxicological Safety of Gamma-Irradiated Salted and Fermented Anchovy Sauce (감마선 조사된 멸치액젓의 유전독성학적 안전성 평가)

  • 육홍선;차보숙;김동호;이주운;변명우
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.7
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    • pp.1192-1200
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    • 2004
  • Gamma irradiations at 5 or 10 kGy were applied to salted and fermented anchovy sauce, for improving the hygiene Quality and evaluating the genotoxicological safety. In vitro genotoxicological safety of irradiated sauces was evaluated by Salmonella Typhimurium (TA98, TA100, TAI535 and TAI537) and E. coli WP2 uvrA, reversion assay, SOS chromotest (Escherichia coli PQ37), and chromosome aberration test (Chinese hamster lung fibroblast cells) in the absence or presence of an exogenous metabolizing system (S9 mix). The gamma-irradiated samples were not significantly different from nonirradiated-control for three in vitro tests (p<0.05). :In vivo micronucleus test using ICR mice (male) was not significantly different from the control at p<0.05. The salted and fermented anchovy sauce exposed to 5 or 10 kGy-gamma ray revealed negative results in these three in vitro mutagenetic tests and in vivo micronucleus test upto 50,000 $\mu$g/plate, respectively. The results indicated that 5 or 10 kGy gamma-irradiated salted and fermented anchovy sauces did not show any mutagenicity.

Mutagenic Assessment of Olmesartan Cilexetil by Bacterial Mutation Assay

  • Kim, Ji Won;Ahn, Ilyoung;Ryu, Sung Ha;Jeon, Hong Ryeol;Lee, Bong Sang;Kim, Kyu-Bong
    • Toxicological Research
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    • v.29 no.3
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    • pp.217-219
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    • 2013
  • Hypertension is a serious health problem due to high frequency and concomitant other diseases including cardiovascular and renal dysfunction. Olmesartan cilexetil is a new antihypertensive drug associated with angiotensin II receptor antagonist. This study was conducted to evaluate the mutagenicity of olmesartan cilexetil by bacterial reverse mutation test using Salmonella typhimurium (TA100, TA1535, TA98, and TA1537) and Escherichia coli (WP2 uvrA). At the concentrations of 0, 62, 185, 556, 1667, and 5000 ${\mu}g$/plate, olmesartan cilexetil was negative in both Salmonella typhimurium and Escherichia coli regardless of presence or absence of metabolic activation system (S9 mix). These results demonstrate that olmesartan cilexetil does not induce bacterial reverse mutation.

Single Dose Oral Toxicity and Genotoxicological Safety Study of Ssanghwa-tang Fermented with Lactobacillus acidophyllus (유산균 발효 쌍화탕에 대한 단회 투여 경구 독성 및 유전 독성 연구)

  • Chung, Tae-Ho;Shim, Ki-Shuk;Kim, Dong-Seon;Lee, Jae-Hoon;Ma, Jin-Yeul
    • The Journal of Korean Medicine
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    • v.32 no.1
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    • pp.67-83
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    • 2011
  • Objectives: The purpose of this study was to examine the single dose toxicity with oral administration and genotoxicities of Ssanghwa-tang fermented with Lactobacillus acidophyllus. Materials and Methods: Clinical signs, weight changes, lethal doses$(LD_{50})$, and postmortem evaluation were determined by Globally Harmonized Classification System(GHCS) in a single-dose oral toxicity study. In vitro mammalian chromosomal aberration test was conducted with Ames test by cell proliferation suppression assessment using the cultivated CHO-K1(Chinese hamster ovary fibroblast) origins. Bacterial reversion assay was performed using Salmonella typhimurium (TA98, TA100, TA1535, and TA1537) and Escherichia coli (WP2uvrA). In vivo micronucleus test was performed using ICR mouse bone marrow. Results: No clinical sign was observed and none of the groups with doses up to 2000 mg/kg showed significant acute oral toxicity in the single dose oral administration. None of the sample doses taken during the 6 to 18 hour groups showed significant aberrant metaphases comparing to the negative control group in the in vitro mammalian chromosomal aberration test. No evidence of mutagenicity was seen for Escherichia coli (WP2uvrA) or Salmonella typhimurium (TA98, TA100, TA1535, and TA1537). No significant increase in the frequency of micronuclei was seen in the micronucleus test. Conclusion: These results indicate that the $LD_{50}$ value of Ssanghwa-Tang fermented with Lactobacillus acidophyllus may be over 2000 mg/kg and it have no acute oral toxicity and genotoxicity.