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Functional Cell‐Surface Display of a Lipase‐Specific Chaperone
Author(s) -
Wilhelm Susanne,
Rosenau Frank,
Becker Stefan,
Buest Sebastian,
Hausmann Sascha,
Kolmar Harald,
Jaeger KarlErich
Publication year - 2007
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200600203
Subject(s) - foldase , periplasmic space , chaperone (clinical) , escherichia coli , biochemistry , lipase , directed evolution , chemistry , microbiology and biotechnology , biology , enzyme , groel , gene , mutant , medicine , pathology
Lipases are important enzymes in biotechnology. Extracellular bacterial lipases from Pseudomonads and related species require the assistance of specific chaperones, designated “Lif” proteins ( li pase specific f oldases). Lifs, a unique family of steric chaperones, are anchored to the periplasmic side of the inner membrane where they convert lipases into their active conformation. We have previously shown that the autotransporter protein EstA from P. aeruginosa can be used to direct a variety of proteins to the cell surface of Escherichia coli . Here we demonstrate for the first time the functional cell‐surface display of the Lif chaperone and FACS (fluorescence‐activated cell sorting)‐based analysis of bacterial cells that carried foldase–lipase complexes. The model Lif protein, LipH from P. aeruginosa , was displayed at the surface of E. coli cells. Surface exposed LipH was functional and efficiently refolded chemically denatured lipase. The foldase autodisplay system reported here can be used for a variety of applications including the ultrahigh‐throughput screening of large libraries of foldase variants generated by directed evolution.