A halotolerant thermostable lipase from the marine bacterium Oceanobacillus sp PUMB02 with an ability to disrupt bacterial biofilms
A halotolerant thermostable lipase from the marine bacterium Oceanobacillus sp PUMB02 with an ability to disrupt bacterial biofilms
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DOI：
10.4161/bioe.29898
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发表时间：
2014-09-01
期刊：
影响因子：
4.9
通讯作者：
Selvin, Joseph
中科院分区：
文献类型：
--
作者：
Kiran, George Seghal;Lipton, Anuj Nishanth;Selvin, Joseph
从海洋细菌 Oceanobacillus sp. 中纯化并表征了耐盐耐热脂肪酶。泵02。该脂肪酶在 pH、盐度和温度等多种条件下均表现出高度稳定性。它分别在 30 摄氏度和 pH 8.0 时具有最佳活性，并且在较高温度（50-70 摄氏度）和碱性 pH 值下稳定。根据 SDS-PAGE 和 MALDI-TOF 指纹分析，脂肪酶的分子量约为 31 kDa。海洋杆菌增强脂肪酶生产的条件。 PUMB02 是在响应面法引导的优化中获得的，评估了橄榄油、蔗糖、铬酸钾和 NaCl 等因素，最终达到 58.84 U/ml 的水平。评估了 PUMB02 脂肪酶的生物膜破坏潜力，并与海洋海绵宏基因组衍生的耐盐脂肪酶 Lpc53E1 进行比较。观察到两种脂肪酶对潜在的食品病原体（例如蜡样芽孢杆菌 MTCC1272、李斯特菌）具有良好的生物膜破坏活性。 MTCC1143，沙雷氏菌属。 MTCC4822、大肠杆菌 MTCC443、荧光假单胞菌 MTCC1748 和副溶血弧菌 MTCC459。相差显微镜、扫描电子显微镜和共焦激光扫描显微镜显示出对致病生物膜的非常有效的破坏。这项研究表明，海洋来源的水解酶（例如脂肪酶）可能具有抑制食品加工环境中生物膜形成的潜在用途，并且是海洋杆菌属脂肪酶在生物膜破坏策略中潜在应用的第一份报告。
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 degrees C and pH 8.0 respectively and was stable at higher temperatures (50-70 degrees 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.