Regulation of thymidine metabolism in Escherichia coli K-12: optimal conditions for the assay of 1,5-phosphodeoxyribomutase in ultrasonic extracts.
Assay conditions have been established which allow maximum expression of phosphodeoxyribomutase activity in ultrasonic extracts of Escherichia coli K-12. The enzyme requires 2-mercaptoethanol, manganous ions, and glucose 1,6-diphosphate for optimal activity. When cells are grown in minimal medium with glycerol as the carbon source and supplemented with 10(-3)m thymidine, phosphodeoxyribomutase is induced three- to four-fold. Cell pellets may be frozen for 24 hr before sonic disruption without loss of enzyme activity. The assay is highly reproducible.
D002474:Cell-Free System; D003124:Colorimetry; D003470:Culture Media; D004926:Escherichia coli / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism*; D005615:Freezing; D005990:Glycerol / Q000378:metabolism; D006600:Hexosephosphates / Q000494:pharmacology; D007477:Ions; D008345:Manganese / Q000494:pharmacology; D008623:Mercaptoethanol / Q000494:pharmacology; D010770:Phosphotransferases / Q000378:metabolism*; D013053:Spectrophotometry; D013936:Thymidine / Q000378:metabolism*; D014465:Ultrasonics; D014732:Vibration
D003470:Culture Media; D006600:Hexosephosphates; D007477:Ions; D008345:Manganese; D008623:Mercaptoethanol; D010770:Phosphotransferases; D005990:Glycerol; D013936:Thymidine