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A permeabilized-cell assay for transaminase B activity in Escherichia coli K-12

โœ Scribed by Joseph J. Whittaker; Julius H. Jackson

Publisher
Elsevier Science
Year
1980
Tongue
English
Weight
577 KB
Volume
105
Category
Article
ISSN
0003-2697

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โœฆ Synopsis

The assay for transaminase B (EC 2.6.1.6) activity, developed by D. E. Duggan and J. A. Wechsler (1973, Anal. Biochem. 51,67-79) has been modified to allow for the measurement of activity in Escherichia coli cells made permeable by cetyltrimethylammonium bromide (CETAB). A concentration of 10 mg% CETAB was found to be most effective in treating the cells without having a significant effect on transaminase B activity. Extraction of the dinitrophenylhydrazone of 2-oxoisovalerate by toluene was not affected by the CETAB treatment. We further report that the Na,CO, extraction step is not required to measure color formed by the dinitrophenylhydrazone of 2-oxoisovalerate. This CETAB-treated cell assay is accurate to study transaminase B activity through most of the logarithmic phase of growth of Escherichia coli.

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