• Title/Summary/Keyword: nitrosoguanidine mutation

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Production of ${\beta}-Galactosidase$ with High Transgalactosylation Activity by Bacillus sp. A4442 Mutant (변이주 Bacillus sp. A4442에 의한 갈락토스 전이활성이 높은 ${\beta}-Galactosidase$의 생산)

  • Choi, Kyung-Ho;Yang, Sung-Joon;Kim, Min-Hong;Han, Keum-Soo;Yang, Ji-Won;Jung, Jin;In, Man-Jin
    • Applied Biological Chemistry
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    • v.38 no.6
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    • pp.507-511
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    • 1995
  • In an attempt to improve the productivity of ${\beta}-galactosidase$ from Bacillus sp. A1, which was isolated from soil and has remarkably higher transgalactosylation activity than lactose hydrolysis activity, a chemical mutation procedure using N-methyl-N'-nitro-N-nitrosoguanidine followed by selection was conducted. The final selection, designated as Bacillus sp. A4442, turned out to show a substantially increased enzyme productivity. Catabolite repression by glucose and lactose requirement as an inducer for the enzyme biosynthesis, which were shown in the parent strain, was markedly diminished; instead it was found out that galactose acts as another inducer. Because pH of medium, one of the most important factors for cell growth as well as enzyme production, is closely related with the sugar concentration during culture, it was kept in the optimum range of $6.5{\sim}7.5$; for this the initial glucose concentration was adjusted to be 0.5% which was thereafter maintained by the controlled pumping-in of lactose using the pH-stat technique. By doing so, we were able to increase the productivity of ${\beta}-galactosidase$ with high transgalactosylation activity up to $44\;unit/m{\ell}-broth$.

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Isolation of Constitutive Mutant of xylA Gene in Escherichia coli (대장균 xylA 유전자의 구성적 변이주의 분리)

  • Soh, Jae Hyun;Roh, Dong Hyun;Rhee, In Koo
    • Current Research on Agriculture and Life Sciences
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    • v.11
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    • pp.81-89
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    • 1993
  • In order to isolate a mutant which was constitutively expressed in xylA gene, Pxyl-cat-xylA fusion gene was constructed by the insertion of cat gene between xylA promoter and xylA structural gene in pEX13 contained xylA gene. The expression of cat and xylA gene from transformants of xylA mutant DH77 with plasmid pEXC131 containing Pxyl-cat-xylA fusion gene was induced by the addition of 0.4% xylose to media. This results indicated that cat and xylA gene were expressed under control of xylA promoter the presence of xylR gene. We have also isolated constitutive mutant plasmid pEXC131-39 from pEXC131 by trementment with N-methyl-N'-nitro-N-nitrosoguanidine(NTG). cat and xylA gene from pEXC131-39 were constitutively expressed without induction of xylose regardless of xylR gene. Transformants of xylR mutant DH60 with pEXC131-39 also expressed chloramphenicol resistances and xylose isomerase without induction of xylose. This result shows that mutation in region of xylA promoter might make it possible to be constitutively expressed.

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Inhibitory Effects of Poturaca oleracea L. Extract on the Mutagenicity of Various Mutagen (각종 변이원에 대한 쇠비름 추출물의 돌연변이 억제 효과)

  • 최근표;정성원;김은정;함승시
    • Journal of the East Asian Society of Dietary Life
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    • v.7 no.4
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    • pp.527-537
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    • 1997
  • This study was performed to determine the effects of antimutagenicity of Porturaca oleracea L. in Korea. In Ames test, the ethanol extract of Poturaca oleracea L. inhibited mutagenic activity of N-methyl-N'-nitro-N-nitrosoguanidine(MNNG), , 4-nitroquinoline-1-oxide(4NQO), benzo($\alpha$)pyrene (B($\alpha$)P) and 3-amino-1,4-dimethyl-5H-pyrido-(4,3-b)indole (Trp-P-1) in Salmonella typhimurium TA98 and TA100. But hot-water extract Poturaca oleracea L. only Inhibited mutagenic activity of MNNG in Salmonella typhimurium TA100, On 4NQO, the ethanol extract 100-1,600$\mu\textrm{g}$/plate of Porturaca oleracea L. showed a slight inhibitory effect of 13-48%, 4-47% in TA98 and TA100, respectively, but on MNNG, it showed higher inhibitory effect of 6-86% in TA100, And the treatment of 1,600$\mu\textrm{g}$/plate of ethanol extract of Porturacea L. had strong antimutagenicity with 74-87% inhibition against TA98 and TA100 induced by B(a)P and with 85-93% inhibition against TA98 and TA100 induced by Trp-P-1. The ethanol extract was fractionated with ether. chloroform, ethylacetate, butanol and water. Among them, most of the fraction except water fraction showed strong antimutagenicity effects against mutation induced by 4-NQO, MNNG, B(a)P and Trp-P-1. Chloroform fraction had strong antimutagenicity with 91% inhibition against TA100 induced by MNNG, diethyl etherfraction had strong antimutagenicity with 92%, 98% inhibition against TA98 and TA100 induced by 4NQO, Chloroform fraction had strong antimutagenicity with 97% inhibition against TA100 induced by B (a)P and diethyl etherfraction had strong antimutagenicity with 98% inhibition against both strain Induced by Trp-P-1, respectively.

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Bacillus coagulans OFR17 Strain Resistant to Rifampicin and Ofloxacin (리팜피신과 오플록사신에 내성인 Bacillus coagulans OFR17 균주)

  • Kim, Eun-Ah;Oh, Tae-Kwon;Choi, Keum-Hwa;Lee, Jin-Hee;Baek, Moon-Chang;Kim, Byong-Kak;Choi, Eung-Chil
    • YAKHAK HOEJI
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    • v.41 no.4
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    • pp.450-455
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    • 1997
  • The preparation of Bacillus coagulans is used as a therapeutics for human intestinal disorders. However, the bacterium in the preparation is very susceptible to rifampic in and fluoroquinolones. When the preparation is taken with rifampicin or fluoroquinolones, its therapeutic effect can not be expected. So B. coagulans RFR17 resistant to rifampicin was obtained by treating the parent B. coagulans with N-methyl-N'-nitro-N-nitrosoguanidine. B. coagulans OFR17 was produced by serial passage of B. coagulans RFR17 on agar with 2-fold minimal inhibitory concentration of ofloxacin or ciprofloxacin. B. coagulans OFR17 was resistant to fluoroquinolones up to 16~64 fold higher than that for the original strain. B. coagulans OFR17 also exhibited identical characteristics with the parent strain when they were tested for lactic acid production and growth inhibition of E. coli MB4-01 and Shigella sonnei MB4-10411. From in vitro test, it was also identified that rifampicin and ofloxacin are not inactivated by certain factors of B. coagulans OFR17. Conclusively, B. coagulans OFR17 can be regarded as a promising strain which can be developed as the preparation for the treatment of the intestinal disorders of the tuberculosis patients under rifampicin and ofloxacin therapy.

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Antimutagenic and Cytotoxic Effects of an Ethanol Extract of Buckwheat Sprout (메밀싹 에탄올 추출물의 항돌연변이원성 및 암세포 성장억제 효과)

  • Cui, Cheng-Bi;Lee, Eui-Yong;Ham, Seung-Shi;Lee, Deuk-Sik
    • Applied Biological Chemistry
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    • v.51 no.3
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    • pp.212-218
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    • 2008
  • This study was performed to determine the antimutagenic and anticancer effects of ethanol extract of buckwheat sprout using Ames test and SRB assay, respectively. An ethyl acetate fraction (200 ${\mu}/plate$) from the ethanol extract of buckwheat sprout showed inhibition rate of 80.6% against the mutagenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in Salmonella typhimurium TA100 strain. Also the ethyl acetate fraction (200 ${\mu}/plate$) showed higher antimutagenic activity than other fractions against the mutagenesis induced by 4-nitroquinoline-1-oxide (4NQO) in Salmonella typhimurium TA98 and TA100. In addition, the ethyl acetate fraction (200 ${\mu}/plate$) showed high antimutagenic effect of 80.9% and 85.9% against the mutation of TA98 and TA100 strains induced by 3-amino-1,4-dimethyl-5H-pyrido-(4,3-b)indol (Trp-P-1), respectively. The cytotoxic effects of each solvent fraction from the ethanol extract of buckwheat sprout against human cancer cell lines including lung carcinoma (A549), gastric carcinoma (AGS), breast adenocarcinoma (MCF-7), hepatocellular carcinoma (Hep3B), and colon adenocarcinoma (Colo 205) were investigated. The ethyl acetate fraction of buckwheat sprout ethanol extract at the concentration of 1.0 mg/ml showed strong cytotoxic activities of 70.3, 94.8, 79.6, 82.3, and 73.2% against A549, AGS, MCF-7, Hep3B and Colo 205 cancer cell lines, respectively.

Antioxidative and Antimutagenic Effects of the Ethanol Elrtract from Cordyceps militaris (번데기동충하초(Cordyceps militaris) 에탄을 추출물의 항산화성 및 항돌연변이원성 효과)

  • 김미남;오상화;이득식;함승시
    • Food Science and Preservation
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    • v.8 no.1
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    • pp.109-117
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    • 2001
  • Cordyceps militaris is a parasitic fungus that has been used as a Chinese medicine for the treatment of fatigue, debility, kidney disease, tuberculosis, asthma and cardiac insufficiency etc. This study was carried out to determine the antioxidative and antimutagenic effects of Cordyceps militaris using DPPH free radical donating method and Ames test, respectively. They were extracted with ethanol and then further fractionated to n-hexane, chloroform, ethyl acetate, butanol and water, stepwise. Among five fractions, the EtOAc and BuOH fractions showed the highest electron donating activities, about 2-fold higher than other fractions. In Ames test, most of the extracts had strong antimutagenic effects against the mutagenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG), 4-nitroquinoline-1-oxide(4NQO), benzo($\alpha$)pyrene(B($\alpha$)P) and 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indol (Trp-P-1). The EtOH extracts of C. militaris (200 $\mu\textrm{g}$/plate) showed 62.8%, 74.4% and 67.2% inhibitory effects on the mutagenesis induced by 4NQO, B($\alpha$)P and Trp-P-1, respectively, against TA98 strain, whereas 78.1%, 78.6%, 78.6% and 82.7% inhibition were observed on the mutagenesis induced by MNNG, 4NQO, B($\alpha$)P and Trp-P-1, respectively, against TA100 strain. Especially, the BuOH fraction showed the highest antimutagenic effects against mutation induced by MNNG.

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Antimutagenic and Cytotoxic Effects of Kochujang Extracts Added Deep Sea Water Salt and Sea Tangle (해양심층수염 및 다시마분말 첨가 고추장추출물의 항돌연변이성 및 암세포 성장억제효과)

  • Ham, Seung-Shi;Choi, Hyun-Jin;Kim, Soo-Hyun;Oh, Hyun-Taek;Chung, Mi-Ja
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.4
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    • pp.410-415
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    • 2008
  • This study was performed to observe the antimutagenic and cytotoxic activities of methanol extract of kochujang added with sea tangle and deep sea water salts (SDK) and kochujang added with sea tangle (SK) using the Ames test and SRB assay, respectively. The direct antimutagenic effect of SDK and SK methanol extracts were examined by Ames test using Salmonella Typhimurium TA98 and TA100. In the Ames test, methanol extract of SDK and SK alone did not exhibit mutagenicity and most of the samples showed high antimutagenic effects against mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 4-nitroquinoline-1-oxide (4NQO). Methanol extract of SDK ($200{\mu}g$/plate) showed approximately 71.4% inhibitory effect on the mutagenesis induced by 4NQO against TA98 strain; whereas 56.1% and 83.6% inhibitions were observed on the mutagenensis induced by 4NQO and MNNG against TA100 strain. The cytotoxic effects of SDK and SK increased with increasing sample concentration against human cervical adenocarcinoma (HeLa), human hepatocellular carcinoma (Hep3B), human breast adenocarcinoma (MCF-7), human stomach adenocarcinoma (AGS), and human lung carcinoma (A549). The SDK at the concentration of 1 mg/ml showed cytotoxicities of 61.5%, 61.3%, 51.4%, 57.9% and 77.7% against HeLa, Hep3B, MCF-7, AGS and A549, respectively. In contrast 1 mg/ml treatment of SDK and SK methanol extract had only $2{\sim}38%$ cytotoxicity on human transformed primary embryonal kidney cell (293).

Antimutagenic and Cytotoxicity Effects of Agaricus blazei Murill Extracts (아가리쿠스버섯(Agaricus blazei Murill) 추출물의 항돌연변이원성 및 세포독성 효과)

  • Ji, Jeong-Hwan;Kim, Mi-Nam;Choi, Kun-Pyo;Chung, Cha-Kwon;Ham, Seung-Shi
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1371-1378
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    • 2000
  • This study was performed to determine the antimutagenic and cytotoxic effect of Agaricus blazei Murill methanol extract on Salmonella typhimurium TA98, TA100 and human cancer cell lines using Ames test and cytotoxicity assay, respectively. In Ames test, methanol extract from A. blazei Murill did not exhibit any mutagenicity and most of the samples showed high antimutagenic effects against mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG), 4-nitroquinoline-1-oxide(4NQO), 3-amino-1,4-dimethyl-5H-pyrido [4,3-b] indol(Trp-P-1) and $benzo({\alpha})pyrene(B({\alpha})P)$. The methanol extracts of A. blazei Murill$(200\;{\mu}g/plate)$ showed approximately 92.4%, 81.9% and 83.4% inhibitory effect on the mutagenesis induced by 4NQO, Trp-P-1 and $B({\alpha})P$ against TA98 strain, whereas 87.3%, 94.7%. 92.3% and 89.9% inhibitions were observed on the mutagenesis induced by MNNG, 4NQO, Trp-P-1 and $B({\alpha})P$ against TA100 strain. The solvent fractions of methanol extracts from A. blazei Murill except water fraction showed high antimutagenic effects of $70{\sim}90%$ against mutation induced by MNNG, 4NQO. Trp-P-1 and $B({\alpha})P$. In anticancer effects of A. blazei Murill extract and fraction against cancer cell lines including human breast adenocarcinoma(MCF7), human lung carcinoma(A549), human fibrosarcoma(HT1080), human hepatocellular carcinoma(Hep3B), human epitheloid carcinoma(HeLa), human gastric carcinoma(KATO III) and human chronic myelogenous leukemia(K562) were investigated. The treatment of 1 mg/mL A. blazei Murill extracts had the highest cytotoxicity with 91.9% against HeLa, followed by KATO III(88.7%), A549(86.5%) and Hpe3B(84.3%). Whereas 1 mg/mL treatment of A. blazei Murill extracts had only $10{\sim}40%$ cytotoxicity on human normal liver cell (WRL68).

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Antimutagenic and Antitumor Effects of Codonopsis lanceolata Extracts (더덕 추출물의 항돌연변이 및 항종양 효과)

  • Kim, Soo-Hyun;Choi, Hyun-Jin;Chung, Mi-Ja;Cui, Cheng-Bi;Ham, Seung-Shi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.10
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    • pp.1295-1301
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    • 2009
  • This study was carried out to investigate the mutagenic, antimutagenic, cytotoxicity and antitumor effect of Codonopsis lanceolata (CL). CL was extracted with 70% ethanol and then further fractionated to hexane, chloroform, ethyl acetate, butanol and water. Antimutagenic, cytotoxicity and antitumor effects of CL extracts were measured by using Ames test, SRB method, and the tumor growth inhibition test. CL extracts did not show any mutagenicity in the Ames test; however, 70% ethanol extracts and its fractions had strong antimutagenic effects against mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 4-nitroquinoline-1-oxide (4NQO). The ethyl acetate fraction of CL (200 ${\mu}g$/plate) showed approximately 72.1% inhibitory effect on the mutagenesis induced by 4NQO against TA98 strain, whereas 69.6% and 67.0% inhibitions were observed on the mutagenesis induced by MNNG and 4NQO against TA100 strain. In anticancer effects, the cytotoxicity of CL extract and its fractions against cancer cell lines including human cervical adenocarcinoma (HeLa), human hepatocellular carcinoma (HepG2), human breast adenocarcinoma (MCF-7), human lung carcinoma (A549) and transformed primary human embryo kidney (293) were investigated. The treatment of 1 mg/mL CL ethyl acetate fraction had the highest cytotoxicity of 74.5%, 70.7% and 80.3% against HeLa, MCF-7 and A549 cells, respectively. In contrast, the extract and its fractions showed only 2$\sim$31% cytotoxicity for a normal human kidney cell line (293). In vivo anticancer effect of CL extract was tested using Balb/c mice transplanted sarcoma-180 cells. CL ethyl acetate fraction showed the highest inhibition rate of 56.4% at the 50 mg/kg concentration.

Antimutagenic and Antitumor Effects of Adenophora triphylla Extracts (잔대 추출물들의 항돌연변이 및 항종양 효과)

  • Ham, Young-An;Choi, Hyun-Jin;Kim, Soo-Hyun;Chung, Mi-Ja;Ham, Seung-Shi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.1
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    • pp.25-31
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    • 2009
  • This study was carried out to investigate the mutagenic, antimutagenic, cytotoxicity and antitumor effects of Adenophora triphylla (AT). AT was extracted with 70% ethanol and then further fractionated to hexane, chloroform, ethyl acetate, butanol and water. Antimutagenic, cytotoxicity and antitumor effects of AT extracts were measured by using Ames test, SRB method, and the tumor growth inhibition test. AT extracts did not show any mutagenicity in the Ames test; however, 70% ethanol extracts and its fractions had strong antimutagenic effects against mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 4-nitroquinoline-1-oxide (4NQO). The ethyl acetate fraction of AT (200 ${\mu}g$/plate) showed approximately 66.5% inhibitory effect on the mutagenesis induced by 4NQO against TA98 strain, whereas 83.3% and 75.1% inhibitions were observed on the mutagenesis induced by MNNG and 4NQO against TA100 strain. In anticancer effects, the cytotoxicity of AT extract and its fractions against cancer cell lines including human cervical adenocarcinoma (HeLa), human hepatocellular carcinoma (Hep3B), human breast adenocarcinoma (MCF-7), human gastric carcinoma (AGS), human lung carcinoma (A549) and transformed primary human embryo kidney (293) were investigated. The treatment of 1 mg/mL AT ethyl acetate faction had the highest cytotoxicity of 79.9%, 74.9%, 66.0%, 71.0% and 74.3% against HeLa, Hep3B, MCF-7, AGS and A549 cells, respectively. In contrast, the extract and its fractions showed only $3{\sim}36%$ cytotoxicity for a normal human kidney cell line (293). In vivo anti-cancer effect of Adenophora triphylla extract was tested using Balb/c mice transplanted sarcoma-180 cells. Adenophora triphylla ethyl acetate fraction showed the highest inhibition rate of 37.2% at the 50 mg/kg concentration.