DOI QR코드

DOI QR Code

Pseudomonas oleovorans의 pyrimidine nucleotide N-ribosidase의 생성 최적조건

Optimization of Culture Conditions for the Production of Pyrimidine Nucleotide N-Ribosidase from Pseudomonas oleovorans

  • 발행 : 2004.08.01

초록

Pyrimidine nucleoade N-ribosidase (pyrimidine 5'-nucleotide phosphoribo (deoxyribo) hydrolase/ pyrimidine 5'-nucleotide nucleosidase, EC 3.2.2.10)는 CMP와 UMP를 직접 분해하여 cytosine과 uracil를 생성하는 가수분해효소이다. 본 연구에서는 Pseudomonas oleovorans ATCC 8062의 생육과 pyrimidine nu-cleotide N-ribosidase생성에 미치는 탄소원과 질소원의 영향 및 생육인자 등에 대하여 검토했다. 효소생성의 최적배양조건은 2% fumarate, 1.5% peptone, 5% corn steep liquor (CSL)과 1% ammonium chloride의 배지조성(초기 pH 7.0)으로 $28^{\circ}C$, 48시간 진탕 배양이 양호했다. 효소의 활성은 생육이 최대에 도달하는 정지기 후기에 최대에 도달하며, 그 이후부터 급속히 불활성화 되었다. P. oleovorans의 pyrimidine nucleocde N-ribosidase는 UMP에 의하여 유도 생성되지 않으므로 구성효소이며, 내생효소였다.

Pyrimidine nucleotide N-ribosidase (pyrimidine 5'-nucleotide phosphoribo (deoxyribo) hydrolase/pyrimidine 5'-nucleoude nucleosidase, EC 3.2.2.10) directly catalyzes pyrimidine 5'-nucleotide to pyrimidine base and ribose (deoxyribo) 5-phosphate. In order to clarify the best nutritional conditions for the growth and the pyrimidine nucleotide N-ribosidase production of Pseudomonas oleovorans ATCC 8062 the effects of various nutrients such as different carbon and nitrogen sources were studied. For the both the growth and the enzyme production, 2% fumarate, 1.5% peptone, 5% corn steep liquor (CSL) and 1% ammonium chloride were excellent carbon and nitrogen sources, respectively. Optimum pH, temperature, and cultivation time for the enzyme production were 7.0, $28^{\circ}C$, and 48 h, respectively. The pyrimidine nucleotide N-ribosidase of P. oleovorans ATCC 8062 was not induced by UMP and its derivatives, and was constitutive enzyme.

키워드

참고문헌

  1. Agarwal, D. P., G. G. Sanwal and P. S. Krishnan. 1963. Improved paper-chromatographic separation of sugar phosphates by using borate-impregnated paper. Analyst. 88, 969-970 https://doi.org/10.1039/an9638800969
  2. Bankel, L., G. Lindstedt and S. Lindstedt. 1976. On the metabolic role of thymidine 2'-hydroxylase (thymidine, 2-oxoglutarate : oxygen oxidoreductase, EC 1.14.11.3) in Nurospora crassa. FEBS Lett. 71, 147-153 https://doi.org/10.1016/0014-5793(76)80918-0
  3. Imada, A., M. Kuno and S. Igarasi. 1967. Degradation of pyrimidine nucleotides by enzyme systems of Streptomyces. I. Ribose-5-phosphate formation from pyrimidine nucleotides. J. Gen. Appl. Microbiol. 13, 255-265 https://doi.org/10.2323/jgam.13.255
  4. Imada, A. 1967. Degradation of pyrimidine nucleotides by enzyme systems of Streptomyces. II. Pyrimidine 5'-nucleotide phosphoribo (deoxyribo) hydrolase of Streptomyces virginiae. J. Gen. Appl. Microbiol. 13, 267-278 https://doi.org/10.2323/jgam.13.267
  5. Kaltwasser, H. and J. Kramer. 1968. Verwertung von cytosine und uracil durch Hydrogenomonas facilis und Hydrogenomonas H 16. Archiv. fur Mikrobiolgie. 60, 172-181 https://doi.org/10.1007/BF00407642
  6. Lowry, O. H., N. J. Rosebrough, A. L. Farr and R. J. Randall. 1951. Protein measurement with the follin phenol reagent. J. Biol. Chem. 193, 265-275
  7. Neuhard, J. 1968. Pyrimidine nucleotide methabolism and pathways of thymidine triphosphate biosynthesis in Salmonella typhimurium. J. Bacteriol. 96, 1519-1527
  8. Sakai, T., T. Watanabe and I. Chibata. 1968. Metabolism of pyrimidine nucleotides in bacteria. J. Ferm. Technol. 46, 202-213
  9. Sakai, T., T. Watanabe and I. Chibata. 1971. Metabolism of pyrimidine nucleotides in bacteria, (II) Studies on the regulation system of the degradation of nucleotides in Pseudomonas oleovorans. J. Ferm. Technol. 49, 488-498
  10. Sakai, T., T. Watanabe and I. Chibata. 1971. Metabolism of pyrimidine nucleotides in bacteria, (III) enzymatic production of ribose-5-phosphate from uridine-5-monophosphate by Pseudomonas oleovorans. Appl. Microbiol. 22, 1085-1090
  11. Sakai, T., T. Yu and S. Omata. 1976. Distribution of enzymes related to cytidine degradation in bacteria. Agric. Biol. Chem. 40, 1893-1895 https://doi.org/10.1271/bbb1961.40.1893
  12. Somogyi, M. 1952. Notes on sugar determination. J. Biol. Chem. 195, 19-23
  13. West, T. P. and G. A. O'Dpnovan. 1982. Repression of cytosine deaminase by pyrimidines in Salmonella typimurium. J. Bacteriol. 149, 1171-1174
  14. Yu, Tae Shick. 1976. Nutritional and cultural characteristics of cytosine deaminase from Pseudomonase aureofaciens. J. Inst. Nat. Sci. Keimyung university. 3, 97-106