Purification of Materials Produced by Amylocolatosis sp. and Anticancer Effect in Oral Cancer Model

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(1)Journal of Dental Hygiene Science Vol. 3, No. 1, pp. 11~ 14 (2003). Amylocolatosis sp..

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(3) . . 1. 2. Purification of Materials Produced by Amylocolatosis sp. and Anticancer Effect in Oral Cancer Model †. Jung Kim , Young-Min Park1 and Do-Seon Lim2 Department of Dental Hygiene, Suwon Women's College, Suwon City, Kyunggi-do 441-748, Korea 1 Department of Dental Hygiene, Yeojoo Institute of Technology(YIT) Kyo-ri 454-5, Yeojoo-up, Yeojoo-gun, Kyunggi-do 880-705, Korea 2 Department of Dental Hygiene, Seoul Health College, Sungnam City, Kyunggi-do 461-713, Korea. A methylotrophic Actinomycetes strain, which produce the anti-oral cancer activity compound, was isolated from soil and estimated as Amylocolatosis sp. based on taxonomic studies. A methanol didn't have influence on the production of the anticancer compounds. These compound were isolated by ethylacetate extract, silica gel column chromatography, sephadex LH-20 column and reverse phase HPLC. The compounds were very stable under heat (121oC), acid(pH 2.0) and alkali(pH 11.0) treatment. The cytotoxic effect of isolated anticancer compounds on various cancer cell lines such as A549, SNU-1, KB, L1210, and Sarcoma 180 was investigated by MTT assay method. And these produced compounds also showed the broad antimicrobial spectrum to test strains such as bacteria and yeast.

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(23) 45 A549(lung carcinoma, human), SNU-1. (stomach, adenocarcinoma, human), KB(mouth, carcinoma, human), L1210(lymphocyte, leukemia, mouse), Sarcoma 180 (sarcoma, mouse) . RPMI1640(GibcoBRL Co.) 10% fetal calf serum(Hyclone Co.) 100 units penicillin-streptomycin(Gibco BRL Co.) 37oC, 5% CO2, 100% relative humidity . 2~6 , monolayer 0.05% trypsin-EDTA cell dissociation .. 8IÂ? #@L" 8I & z üi z üi7 ý~ L" 8IÂ }H £ø" 2 )LY 0 8IÂ @þ6 L". Procedure for purification method of anti-oral cancer activity materials by Amylocolatosis sp...

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(43) & FR ©6 û{=".. lacetate magnesium sulfate , ethanol , 10 overnight water 4 oC (12,000 rpm, 30 ) methylene chloride silica gel 60 column(φ35 600 mm, Merck Co.), methylene chloride-ethanol(96: 4) sephadex LH-20 column(φ15 300 mm, Pharmacia LKB) 100% methanol HPLC (Waters, ODS) . 2. Compound Gram(+) (6 ), Gram(−) (11 ), (2 ), (1 ) . 3. ,. . 1. Faber E: The multistep nature of cancer development. Cancer Res 44: 4217-4225, 1984. 2. Slaughter DP, Southwick HW, Smejkal W: Field cancerization in oral stratified squamous epithelium. Clinical implication of multicentric origin. Cancer 6: 963-968, 1953. 3. Silverman S Jr, Gorsky M, Lozada NF: Oral leukoplakia and malignant transformation, Cancer 53: 563-568, 1984. 4. Masayuki H, Nomomura H: A new method for intensive isolation of Actinomycetes from soil. Actinomycetol 3: 95-104, 1989. 5. Hayakawa M, Nonomura H: Humic acid-vitamin agar, a new medium for the selective isolation of soil Actinomycetes. J Fermet Technol 65: 501-509, 1987. 6. Kim HS: Isolation and production of antifungal compound from methylotrophic actinomycetes strain M-17. N Inst Nat Sci 17: 75-84, 1977. 7. Miyazak SS, Toki I, Izumi Y, Yamada H: Purification and characterization of methanol dehydrogenase of a serine producing methylotroph, Hyphomicrobium methylvorum. J Ferment Technol 65: 371-377, 1987. 8. Yoon BD, Ueno M, Tami Y: Improvement of cytochrome c production by Glycin and Glycine Analog resistant mutants of Methyllomonas sp. J Ferment Technol 52: 917-920, 1987. 9. Kato N, Tsuji K, Tani Y, Ogata K: A Methanol-utilizing Actinomycetes. J Ferment Technol 52: 917-920. 1974. 10. Shibuya H, Amagasa T, Seto K, Ishibashi K, Horiuchi J, Suzuki S: Leukoplakia-associated multiple carcinomas in patients with tongue carcinoma. Cancer 57: 843, 1986. 11. Robinson E, Neugut Al, Murray T, Rennert G: A comparison of the clinical characterization of first and second primary head and neck cancers. A population based study. Cancer 68: 189, 1991. 12. Pinborg JJ, Renstrup G, Jolst O, Roed-Petersen B: Studies in oral leukoplakia: a preliminary report on the period prevalence of malignant transformation in leukoplakia based on a follow-up study of 248 patient, J Am Dent Assoc 76: 767-771, 1968. 13. Silverman S Jr, Gorsky M, Lozada F: Oral leukoplakia and malignant transformation. Cancer 53: 563-568, 1984. (Received May 21, 2003; Accepted June 16, 2003).

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