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Sagrario Arias, Monica Bassas-Galia, Gabriella Molinari and Kenneth N. Timmis Tight coupling of polymerization and depolymerization of polyhydroxyalkanoates ensures efficient management of carbon resources in Pseudomonas putida Microbial Biotechnology 6

Version of Record online: 28 FEB 2013 | DOI: 10.1111/1751-7915.12040

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In order to obtain insights into the role of polyhydroxyalkanoate (PHA) depolymerase (PhaZ) and its relationship to a PHA polymerase (PhaC2) in the carbon management activity of Pseudomonas putida strain U, PHA polymerase hyperexpression and PHA depolymerase knockout mutants were created and their phenotypes analyzed. Hyperexpression of PhaC2 led to accumulation of higher amounts of PHA (44 %wt) than in the wild type strain (24 %wt) after 24 h of cultivation, which then returned to wild-type levels by 48 h, as a result of elevated depolymerization. The phaZ mutant, however, accumulated higher levels of PHA than the parental strain (62 %wt), which were maintained for at least 96 h. Transcriptional analysis of the pha cluster by RT-PCR revealed regulatory coupling of PHA polymerase and depolymerase activities that act in synergy, and in concert with other PHA proteins, to provide dynamic PHA granule synthesis and remodeling that rapidly and sensitively respond to changes in availability of carbon and the physiological-metabolic needs of the cell, to ensure optimal carbon resource management.

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