A generic system for the Escherichia coli cell-surface display of lipolytic enzymes.
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.
Becker
Stefan
S
Abteilung für Molekulare Genetik und Präparative Molekularbiologie, Institut für Mikrobiologie und Genetik, Georg-August-Universität Göttingen, Grisebachstrasse 8, D-37077 Göttingen, Germany.
Theile
Sebastian
S
Heppeler
Nele
N
Michalczyk
Anja
A
Wentzel
Alexander
A
Wilhelm
Susanne
S
Jaeger
Karl-Erich
KE
Kolmar
Harald
H
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics; D008049:Lipase / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D011550:Pseudomonas aeruginosa; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012706:Serratia marcescens / Q000201:enzymology / Q000235:genetics
D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D002265:Carboxylic Ester Hydrolases; C098953:EstA protein, bacteria; D008049:Lipase