Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, 8 Medical Drive, Singapore 117596, Singapore.
NUS Synthetic Biology for Clinical and Technological Innovation, Centre for Life Sciences, National University of Singapore, 14 Medical Drive, Singapore 117456, Singapore.
Molecules. 2021 Mar 11;26(6):1542. doi: 10.3390/molecules26061542.
Lipases and esterases are important catalysts with wide varieties of industrial applications. Although many methods have been established for detecting their activities, a simple and sensitive approach for picogram detection of lipolytic enzyme quantity is still highly desirable. Here we report a lipase detection assay which is 1000-fold more sensitive than previously reported methods. Our assay enables the detection of as low as 5 pg and 180 pg of lipolytic activity by direct spotting and zymography, respectively. Furthermore, we demonstrated that the detection sensitivity was adjustable by varying the buffering capacity, which allows for screening of both high and low abundance lipolytic enzymes. Coupled with liquid chromatography-mass spectrometry, our method provides a useful tool for sensitive detection and identification of lipolytic enzymes.
脂肪酶和酯酶是具有广泛工业应用的重要催化剂。尽管已经建立了许多方法来检测它们的活性，但仍然非常需要一种简单、灵敏的方法来检测痕量的脂肪酶数量。在这里，我们报告了一种比以前报道的方法灵敏 1000 倍的脂肪酶检测方法。我们的测定方法可以直接点样和酶谱法检测低至 5pg 和 180pg 的脂肪酶活性。此外，我们证明通过改变缓冲能力可以调节检测灵敏度，这允许筛选高丰度和低丰度的脂肪酶。与液相色谱-质谱联用，我们的方法为灵敏检测和鉴定脂肪酶提供了一种有用的工具。
