Pyrimidine Ribonucleoside Salvage Metabolism in Pseudomonas Putida ATCC 17536

Sara Fatima

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Chemistry

Date of Award

8-14-2024

Abstract

The study aimed to investigate the pyrimidine ribonucleoside salvage metabolism in Pseudomonas putida ATCC 17536. Specifically, the goal was to assess the enzyme activities involved in the pyrimidine salvage pathway in ATCC 17536 cells cultured in minimal media containing 0.4% succinate or glucose, supplemented with 0.2% pyrimidine bases and ribonucleosides as potential nitrogen sources. The objective was to measure the activities of cytosine deaminase, uridine phosphorylase, and nucleoside hydrolase in extracts from ATCC 17536 cells grown under specific carbon and nitrogen conditions. The bacterial cells were cultured in minimal media containing 0.4% glucose or sodium succinate as the carbon source, with various 0.2% pyrimidine bases as nitrogen sources. ATCC 17536 Cells in the late exponential phase were collected, disrupted using ultrasound, and the cell extract was centrifuged which was followed by overnight dialysis at 4°C. Results indicated that the carbon source influenced the pyrimidine ribonucleoside salvage pathway in P. putida ATCC 17536, with higher activities being observed in the succinate medium compared to the glucose medium. Additionally, the impact of adding pyrimidine bases to the culture medium was investigated, which revealed differential effects on salvage enzymes depending on the carbon source. In summary, this investigation sheds light on the regulation of pyrimidine base and ribonucleoside salvage metabolism in P. putida ATCC 17536 demonstrating the influence of carbon and nitrogen sources on enzyme synthesis, which fills a void in the pyrimidine biochemistry of P. putida cells.

Advisor

Thomas West

Subject Categories

Chemistry | Physical Sciences and Mathematics

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