An AGT-based protein-tag system for the labelling and surface immobilization of enzymes on E. coli outer membrane

Journal of Enzyme Inhibition and Medicinal Chemistry

Sonia Del Prete¹  Clemente Capasso¹  Giuseppe Perugino¹  Rosanna Mattossovich¹  Vincenzo Carginale¹  Rosa Merlo¹  Anna Valenti¹  Claudiu T. Supuran² 

[1] Department of Biology Agriculture and Food Sciences, Institute of Bioscience and BioResources – National Research Council of Italy, Naples, Italy;Neurofarba Department, University of Florence, Polo Scientifico, Sesto Fiorentino Firenze, Ital;

关键词: Carbonic anhydrase;    β;    protein-tag;    ice nucleation protein;    enzyme immobilisation;   

DOI  :  10.1080/14756366.2018.1559161

来源: publisher

【 摘 要 】

The use of natural systems, such as outer membrane protein A (OmpA), phosphoporin E (PhoE), ice nucleation protein (INP), etc., has been proved very useful for the surface exposure of proteins on the outer membrane of Gram-negative bacteria. These strategies have the clear advantage of unifying in a one-step the production, the purification and the in vivo immobilisation of proteins/biocatalysts onto a specific biological support. Here, we introduce the novel Anchoring-and-Self-Labelling-protein-tag (ASLtag), which allows the in vivo immobilisation of enzymes on E. coli surface and the labelling of the neosynthesised proteins with the engineered alkylguanine-DNA-alkyl-transferase (H5) from Sulfolobus solfataricus. Our results demonstrated that this tag enhanced the overexpression of thermostable enzymes, such as the carbonic anhydrase (SspCA) from Sulfurihydrogenibium yellowstonense and the β-glycoside hydrolase (SsβGly) from S. solfataricus, without affecting their folding and catalytic activity, proposing a new tool for the improvement in the utilisation of biocatalysts of biotechnological interest.

【 授权许可】

CC BY