Controlled lid-opening in Thermomyces lanuginosus lipase- An engineered switch for studying lipase function

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• 作者：Jakob Skjold-Jø ; rgensen^a ; ^b ; Jesper Vind^b ; Olga V. Moroz^c ; Elena Blagova^c ; Vikram K. Bhatia^b ; Allan Svendsen^b ; Keith S. Wilson^c ; ^{keith.wilson@york.ac.uk} ; Morten J. Bjerrum^a ; ^{mobj@chem.ku.dk}
• 关键词：CMC ; critical micelle concentration ; pNP ; para-nitrophenol ; QCM-D ; quartz crystal microbalance with dissipation monitoring ; TCEP ; Tris (2-Carboxyethyl) phosphine
• 刊名：Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics
• 出版年：2017
• 出版时间：January 2017
• 年：2017
• 卷：1865
• 期：1
• 页码：20-27
• 全文大小：845 K
• 卷排序：1865

文摘

Rational design and expression of a lipase variant containing a switch controlling opening and closing of the lid. The closed form of the lipase displayed very low enzymatic activity and interfacial binding compared to wild-type TlL. Release of the lock on the lid by reducing agent restored enzymatic activity. The crystal structure shows formation of a disulfide bond in the locked form which causes strained closure of the lid-domain. The crystal structure reveal sites of interest for rational design of lipases that activate under controlled conditions.