Year: 2016
Type: dissertation
Abstract: During chronic infection, Pseudomonas aeruginosa generate alginate-containing biofilms in the lungs of cystic fibrosis patients. Mature bacterial alginate is a heteropolymer of ι-L-guluronic acid and partially acetylated β-D-mannuronic acid. P. aeruginosa alginate biosynthesis requires thirteen proteins, including an alginate lyase, AlgLPa, which has been proposed to have a bi-functional role, both as a lyase and as an integral part of the cell envelope biosynthetic complex. How AlgLPa performs these functions is currently poorly understood. We have used an in vitro structure-function approach to explore the substrate recognition and catalytic mechanism of AlgLPa, and in vivo complementation to explore the role of its activity in alginate biosynthesis. The results presented suggest that AlgLPa is an endo-acting lyase with at least six subsites, and that the loop containing K66 is critical for substrate binding and/or catalysis. The complementation data supports the hypothesis that AlgLPa assembles into the cell envelope biosynthetic complex.
Source: TSpace (University of Toronto)
Author: Steven Wong
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W3088978835