Priyanka Priyanka, Kinsella Gemma, Henehan Gary T, Ryan Barry J
Dublin Institute of Technology, Ireland.
Dublin Institute of Technology, Ireland.
Protein Expr Purif. 2019 Jan;153:121-130. doi: 10.1016/j.pep.2018.08.007. Epub 2018 Aug 22.
The Pseudomonas sp. have been long recognized for their exogenous lipolytic activities yet the genus still contains a lot of unexplored strains. Due to the versatile metabolic machinery and their potential for adaptation to fluctuating environmental conditions Pseudomonas sp. are of great interest for biotechnological applications. In this study, a new extracellularly produced lipolytic enzyme from Pseudomonas sp. (P. reinekei) was purified and characterized. The production of lipase from P. reinekei (H1) was enhanced 10-fold by optimizing the nitrogen source. The 50 kDa H1 lipase was purified using negative and positive mode anion exchange chromatography. The purified lipase was active over a broad pH range (5.0-9.0) and was stable for 24 h at 40 °C. The lipase showed significant stability, and indeed activation, in the presence of organic solvents with log P ≥ 2.0. These features render this lipase of interest as a biocatalyst for applications such as biodiesel production, detergent formulations and biodegradation of oil in the environment.
假单胞菌属因其胞外脂肪分解活性早已为人所知，但该属仍包含许多未被探索的菌株。由于其多样的代谢机制以及适应波动环境条件的潜力，假单胞菌属在生物技术应用方面具有极大的吸引力。在本研究中，一种来自假单胞菌属（雷氏假单胞菌）新的胞外产生的脂肪分解酶被纯化并进行了表征。通过优化氮源，雷氏假单胞菌（H1）脂肪酶的产量提高了10倍。使用正负模式阴离子交换色谱法纯化了50 kDa的H1脂肪酶。纯化后的脂肪酶在较宽的pH范围（5.0 - 9.0）内具有活性，并且在40°C下可稳定24小时。在log P≥2.0的有机溶剂存在下，该脂肪酶表现出显著的稳定性，甚至有激活现象。这些特性使得这种脂肪酶作为生物催化剂可用于生物柴油生产、洗涤剂配方以及环境中石油的生物降解等应用。
