Monitoring transport of a recombinant phosphotriesterase in Streptomyces lividans using a pulse-chase system |
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Authors: | M Sathyamoorthy M K Speedie |
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Institution: | (1) Department of Biomedicinal Chemistry, School of Pharmacy, University of Maryland, 21201 Baltimore, MD, USA |
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Abstract: | A heterologous phosphotriesterase (parathion hydrolase) was previosly shown to be secreted by Streptomyces lividans. To investigate the mechanism of secretion, a system to label the protein and follow its expression and secretion was developed. The recombinant S. lividans was grown first in a defined medium containing 35S]methionine that permitted expression but not secretion. It was then transferred to tryptone/glucose medium with unlabeled methionine for the chase period, during which secretion was observed. The results indicate a relatively slow rate of secretion that is also dependent on the growth medium. |
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