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

Pseudomonas oleovorans의 pyrimidine nucleotide N-ribosidase의 생성 최적조건

  • Published : 2004.08.01

Abstract

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.

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.

Keywords

References

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