TY  - JOUR
AB  - Pseudomonas putida can be used as a host for the autotransporter‐mediated surface display of enzymes (autodisplay), resulting in whole‐cell biocatalysts with recombinant functionalities on their cell envelope. The efficiency of autotransporter‐mediated secretion depends on the N‐terminal signal peptide as well as on the C‐terminal translocator domain of autotransporter fusion proteins. We set out to optimize autodisplay for P. putida as the host bacterium by comparing different signal peptides and translocator domains for the surface display of an esterase. The translocator domain did not have a considerable effect on the activity of the whole‐cell catalysts. In contrast, by using the signal peptide of the P. putida outer membrane protein OprF, the activity was more than 12‐fold enhanced to 638 mU ml−1 OD−1 compared with the signal peptide of V. cholerae CtxB (52 mU ml−1 OD−1). This positive effect was confirmed with a β‐glucosidase as a second example enzyme. Here, cells expressing the protein with N‐terminal OprF signal peptide showed more than fourfold higher β‐glucosidase activity (181 mU ml−1 OD−1) than with the CtxB signal peptide (42 mU ml−1 OD−1). SDS‐PAGE and flow cytometry analyses indicated that the increased activities correlated with an increased amount of recombinant protein in the outer membrane and a higher number of enzymes detectable on the cell surface.
AU  - Tozakidis, Iasson
AU  - Tozakidis, Iasson E. P.
AU  - Tozakidis, Iasson Elias Philippos
AU  - Lüken, Lena M.
AU  - Üffing, Alina
AU  - Meyers, Annika
AU  - Jose, Joachim
DA  - 2019-05-02
DO  - 10.1111/1751-7915.13419
LA  - eng
N1  - Microbial Biotechnology 13 (2019) 1, 176-184
N1  - Finanziert durch den Open-Access-Publikationsfonds der Westfälischen Wilhelms-Universität Münster (WWU Münster)
N1  - Funding Information: Bundesministerium für Bildung und Forschung (031B0002B)
PY  - 2019-05-02
TI  - Improving the autotransporter-based surface display of enzymes in Pseudomonas putida KT2440
UR  - https://nbn-resolving.org/urn:nbn:de:hbz:6-78069385464
Y2  - 2024-11-21T23:35:52
ER  -