A halotolerant thermostable lipase from the marine bacteriumOceanobacillussp. PUMB02 with an ability to disrupt bacterial biofilms

Abstract

A halotolerant thermostable lipase was purified and characterized from the marine bacterium Oceanobacillus sp. PUMB02. This lipase displayed a high degree of stability over a wide range of conditions including pH, salinity, and temperature. It was optimally active at 30 °C and pH 8.0 respectively and was stable at higher temperatures (50-70 °C) and alkaline pH. The molecular mass of the lipase was approximately 31 kDa based on SDS-PAGE and MALDI-ToF fingerprint analysis. Conditions for enhanced production of lipase by Oceanobacillus sp. PUMB02 were attained in response surface method-guided optimization with factors such as olive oil, sucrose, potassium chromate, and NaCl being evaluated, resulting in levels of 58.84 U/ml being achieved. The biofilm disruption potential of the PUMB02 lipase was evaluated and compared with a marine sponge metagenome derived halotolerant lipase Lpc53E1. Good biofilm disruption activity was observed with both lipases against potential food pathogens such as Bacillus cereus MTCC1272, Listeria sp. MTCC1143, Serratia sp. MTCC4822, Escherichia coli MTCC443, Pseudomonas fluorescens MTCC1748, and Vibrio parahemolyticus MTCC459. Phase contrast microscopy, scanning electron microscopy, and confocal laser scanning microscopy showed very effective disruption of pathogenic biofilms. This study reveals that marine derived hydrolytic enzymes such as lipases may have potential utility in inhibiting biofilm formation in a food processing environment and is the first report of the potential application of lipases from the genus Oceanobacillus in biofilm disruption strategies.

Production of enzymes and siderophores by epiphytic bacteria isolated from the marine macroalga Ulva lactuca

“…After the sterilization process, agar nutrient, tributyrin and Triton X-100 (emulsifier) were sonicated in distilled water for 30 min. Isolate colonies were streaked on tributyrin plates and incubated at 30°C for 48 h (Kiran et al 2014). Colonies with activity showed a clear halo formed by the hydrolysis of tributyrin.…”

Biopelículas y persistencia microbiana en la industria alimentaria

“…Como el ADN extracelular a538 es un componente habitual de la matriz de biopelículas microbianas, las enzimas con acción DNasa, solas o combinadas con otras estrategias de saneamiento, pueden facilitar la eliminación de las biopelículas, como se ha demostrado recientemente para C. jejuni (Brown, Hanman, Reuter, Betts y van Vliet, 2015) o L. monocytogenes (Nguyen y Burrows, 2014). Además, las proteasas, como la proteinasa K (Nguyen y Burrows, 2014), las lipasas (Kiran, Lipton, Kennedy, Dobson y Selvin, 2014) o las enzimas que degradan carbohidratos, como la β-glucanasa y la α-amilasa (Araújo et al, 2017), por su actividad lítica sobre otros componentes de la matriz extracelular de las biopelículas, también se han propuesto como posibles candidatos para ser utilizados como herramientas de control.…”

Response Surface Methodology (RSM) for analysing culture conditions of Acidocella facilis strain USBA-GBX-505 and Partial Purification and Biochemical Characterization of Lipase 505 LIP

“…We used the titration method to determine the free fatty acids concentrations, according to Seghal et al (2014) [7]. Briefly, the reaction mixture contained 5% (w/v) of the lipidic substrate (tributyrine, tricapryline and trioleate) in Tris-HCl buffer (50 mM, pH 7.5) and 1% of Triton X-100.…”

“…These enzymes are industrially important due to their capacity to act in environments with a low concentration of water, allowing them to perform esterification, transesterification, aminolytic and acidolytic reactions [4] with high efficiency and stability. The lipases are able to http://ciencias.javeriana.edu.co/investigacion/universitas-scientiarum catalyse enzymatic reactions with efficiency, stability and chemo-, region-and enantioselectivity, as well as not requiring cofactors and high activity levels in organic solvents make them versatile biocatalysers for industrial purposes [5], including food, dairy, agrochemical, pharmaceutical, cosmetic, and detergent production [5,6] due to their unique properties, associated with their broad substrate range [7].…”