Enhancement of the Antibiofilm Activity Against Pseudomonas aeruginosa and Biodiesel Production Using Lipase from Cronobacter dublinensis as a Catalyst
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Abstract
Lipases can convert edible and vegetable oils into biodiesel, a low-toxicity, renewable, biodegradable, and environmentally friendly alternative to traditional fuel. This study investigated the dual functionality of lipase from Cronobacter dublinensis in enhancing antibiofilm activity against Pseudomonas aeruginosa and catalyzing biodiesel production via enzymatic transesterification. Cronobacter dublinensis was isolated from lettuce samples and identified by morphological, biochemical, and molecular methods, with gyrB gene confirmation. Lipase production was screened on oil-supplemented agar, and the gene known as Lip was detected via polymerase chain reaction (PCR). The lipase was then purified using ammonium sulfate precipitation in combination with dialysis and chromatographic techniques. Antibiofilm activity was assessed using a microtiter plate method against Pseudomonas aeruginosa. Biodiesel production was performed via enzymatic transesterification using crude lipase. Four Cronobacter dublinensis isolates were identified from the lettuce samples. Lipase production induced by various oils revealed that olive oil yielded the highest hydrolytic activity, producing zones of 20–50 mm. The Lip gene was detected in all the isolates via PCR (550 bp fragment). Purification through the combined use of ammonium sulfate precipitation and chromatography resulted in a 5.5-times improvement in purity with 58% recovery. Purified lipase significantly inhibited the establishment of biofilms by Pseudomonas aeruginosa, which varied with dosage, showing maximum suppression at a concentration of 200 μg/mL. Enzymatic transesterification using C. dublinensis lipase achieved 38% and 11.2% fatty acid methyl ester yields for processed olive and corn oils, respectively. These findings highlight the dual potential of C. dublinensis lipase in antimicrobial and biofuel applications.
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