| Download | - View author's version: Assay of thymidine phosphorylase in whole cell suspensions of Escherichia coli (PDF, 1.2 MiB)
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| DOI | Resolve DOI: https://doi.org/10.1016/B978-0-08-025385-5.50093-6 |
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| Author | Search for: Brosseau, J. D.; Search for: Khachatourians, G. G.; Search for: Child, J. J.1 |
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| Affiliation | - National Research Council of Canada. Prairie Regional Laboratory
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| Format | Text, Article |
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| Conference | Sixth International Fermentation Symposium, July 20–25, 1980, London, Ontario, Canada |
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| Subject | thymidine; phosphorylase; assay |
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| Abstract | A simple in situ assay for thymidine phosphorylase activity (E.C.2.4.2.4.) in intact E. coli cells is described which requires only standard spectrophotometric equipment. The assay is based upon a characteristic hyperchromic shift for the product thymine under a change from neutral to alkaline conditions. Conversion reactions can therefor be quantitated in terms of the increased absorption at 290 nm following addition of NaOH.
The enzymic activity may be measured in actively growing or non-growing cells. Maximum rates of activity (μmoles thymine·min⁻¹·A₆₀₀⁻¹) have been recorded at pH 5.5 and 47°C for non-growing cells and at pH 6.5 and 37°C for actively growing cells. The assay is sufficiently sensitive and convenient to permit the measurement of thymidine conversion in stirred tank or continuous flow reactors without the need for cell disruption. |
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| Publication date | 1981 |
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| Publisher | Elsevier |
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| In | |
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| Language | English |
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| Peer reviewed | Yes |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction (opens in a new tab) |
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| Record identifier | 3e4f3151-3e69-4373-bdfd-c55fd09bdbf3 |
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| Record created | 2023-11-15 |
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| Record modified | 2025-04-09 |
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