Becker Stefan, Theile Sebastian, Heppeler Nele, Michalczyk Anja, Wentzel Alexander, Wilhelm Susanne, Jaeger Karl-Erich, Kolmar Harald
Abteilung für Molekulare Genetik und Präparative Molekularbiologie, Institut für Mikrobiologie und Genetik, Georg-August-Universität Göttingen, Grisebachstrasse 8, D-37077 Göttingen, Germany.
FEBS Lett. 2005 Feb 14;579(5):1177-82. doi: 10.1016/j.febslet.2004.12.087.
EstA is an outer membrane-anchored esterase from Pseudomonas aeruginosa. An inactive EstA variant was used as an anchoring motif for the Escherichia coli cell-surface display of lipolytic enzymes. Flow cytometry analysis and measurement of lipase activity revealed that Bacillus subtilis lipase LipA, Fusarium solani pisi cutinase and one of the largest lipases presently known, namely Serratia marcescens lipase were all efficiently exported by the EstA autotransporter and also retained their lipolytic activities upon cell surface exposition. EstA provides a useful tool for surface display of lipases including variant libraries generated by directed evolution thereby enabling the identification of novel enzymes with interesting biological and biotechnological ramifications.
EstA是一种来自铜绿假单胞菌的外膜锚定酯酶。一种无活性的EstA变体被用作脂肪分解酶在大肠杆菌细胞表面展示的锚定基序。流式细胞术分析和脂肪酶活性测量表明，枯草芽孢杆菌脂肪酶LipA、茄病镰刀菌角质酶以及目前已知的最大脂肪酶之一，即粘质沙雷氏菌脂肪酶，均通过EstA自转运蛋白有效输出，并且在细胞表面展示时仍保留其脂肪分解活性。EstA为脂肪酶的表面展示提供了一种有用的工具，包括通过定向进化产生的变体文库，从而能够鉴定具有有趣生物学和生物技术意义的新型酶。
