Surface Display of Designer Protein Scaffolds on Genome-Reduced
Strains of Pseudomonas putida

J 2020

Surface Display of Designer Protein Scaffolds on Genome-Reduced Strains of Pseudomonas putida

DVOŘÁK, Pavel; Edward A. BAYER a Víctor DE LORENZO

Základní údaje

Originální název

Surface Display of Designer Protein Scaffolds on Genome-Reduced Strains of Pseudomonas putida

Autoři

DVOŘÁK, Pavel (203 Česká republika, garant, domácí); Edward A. BAYER a Víctor DE LORENZO

Vydání

ACS Synthetic Biology, Washington, DC, American Chemical Society, 2020, 2161-5063

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 5.110

Kód RIV

RIV/00216224:14310/20:00114419

Organizační jednotka

Přírodovědecká fakulta

DOI

https://doi.org/10.1021/acssynbio.0c00276

UT WoS

000582582100014

EID Scopus

2-s2.0-85093538854

Klíčová slova anglicky

pseudomonas putida; cellulosome; designer scaffoldin; surface display; synthetic biology

Štítky

rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 7. 12. 2020 15:24, Mgr. Marie Novosadová Šípková, DiS.

Anotace

V originále

The bacterium Pseudomonas putida KT2440 is gaining considerable interest as a microbial platform for biotechnological valorization of polymeric organic materials, such as lignocellulosic residues or plastics. However, P. putida on its own cannot make much use of such complex substrates, mainly because it lacks an efficient extracellular depolymerizing apparatus. We seek to address this limitation by adopting a recombinant cellulosome strategy for this host. In this work, we report an essential step in this endeavor—a display of designer enzyme-anchoring protein “scaffoldins”, encompassing cohesin binding domains from divergent cellulolytic bacterial species on the P. putida surface. Two P. putida chassis strains, EM42 and EM371, with streamlined genomes and differences in the composition of the outer membrane were employed in this study. Scaffoldin variants were optimally delivered to their surface with one of four tested autotransporter systems (Ag43 from Escherichia coli), and the efficient display was confirmed by extracellular attachment of chimeric beta-glucosidase and fluorescent proteins. Our results not only highlight the value of cell surface engineering for presentation of recombinant proteins on the envelope of Gram-negative bacteria but also pave the way toward designer cellulosome strategies tailored for P. putida.

Návaznosti

GJ19-06511Y, projekt VaV

Název: Ortogonalizace metabolismu sacharidů v bakteriálním šasi Pseudomonas putida EM42 pro ko-utilizaci cukrů z rostlinné biomasy

Investor: Grantová agentura ČR, Ortogonalizace metabolismu sacharidů v bakteriálním šasi Pseudomonas putida EM42 pro ko-utilizaci cukrů z rostlinné biomasy

Citovat

DVOŘÁK, Pavel; Edward A. BAYER a Víctor DE LORENZO. Surface Display of Designer Protein Scaffolds on Genome-Reduced Strains of Pseudomonas putida. ACS Synthetic Biology. Washington, DC: American Chemical Society, 2020, roč. 9, č. 10, s. 2749-2764. ISSN 2161-5063. Dostupné z: https://doi.org/10.1021/acssynbio.0c00276.

@article{1688740,
   author = {Dvořák, Pavel and Bayer, Edward A. and de Lorenzo, Víctor},
   article_location = {Washington, DC},
   article_number = {10},
   doi = {https://doi.org/10.1021/acssynbio.0c00276},
   keywords = {pseudomonas putida; cellulosome; designer scaffoldin; surface display; synthetic biology},
   language = {eng},
   issn = {2161-5063},
   journal = {ACS Synthetic Biology},
   title = {Surface Display of Designer Protein Scaffolds on Genome-Reduced Strains of Pseudomonas putida},
   url = {https://doi.org/10.1021/acssynbio.0c00276},
   volume = {9},
   year = {2020}
}

TY  - JOUR
ID  - 1688740
AU  - Dvořák, Pavel - Bayer, Edward A. - de Lorenzo, Víctor
PY  - 2020
TI  - Surface Display of Designer Protein Scaffolds on Genome-Reduced Strains of Pseudomonas putida
JF  - ACS Synthetic Biology
VL  - 9
IS  - 10
SP  - 2749-2764
EP  - 2749-2764
PB  - American Chemical Society
SN  - 21615063
KW  - pseudomonas putida
KW  - cellulosome
KW  - designer scaffoldin
KW  - surface display
KW  - synthetic biology
UR  - https://doi.org/10.1021/acssynbio.0c00276
L2  - https://doi.org/10.1021/acssynbio.0c00276
N2  - The bacterium Pseudomonas putida KT2440 is gaining considerable interest as a microbial platform for biotechnological valorization of polymeric organic materials, such as lignocellulosic residues or plastics. However, P. putida on its own cannot make much use of such complex substrates, mainly because it lacks an efficient extracellular depolymerizing apparatus. We seek to address this limitation by adopting a recombinant cellulosome strategy for this host. In this work, we report an essential step in this endeavor—a display of designer enzyme-anchoring protein “scaffoldins”, encompassing cohesin binding domains from divergent cellulolytic bacterial species on the P. putida surface. Two P. putida chassis strains, EM42 and EM371, with streamlined genomes and differences in the composition of the outer membrane were employed in this study. Scaffoldin variants were optimally delivered to their surface with one of four tested autotransporter systems (Ag43 from Escherichia coli), and the efficient display was confirmed by extracellular attachment of chimeric beta-glucosidase and fluorescent proteins. Our results not only highlight the value of cell surface engineering for presentation of recombinant proteins on the envelope of Gram-negative bacteria but also pave the way toward designer cellulosome strategies tailored for P. putida.
ER  -

DVOŘÁK, Pavel; Edward A. BAYER a Víctor DE LORENZO. Surface Display of Designer Protein Scaffolds on Genome-Reduced Strains of Pseudomonas putida. \textit{ACS Synthetic Biology}. Washington, DC: American Chemical Society, 2020, roč.~9, č.~10, s.~2749-2764. ISSN~2161-5063. Dostupné z: https://doi.org/10.1021/acssynbio.0c00276.

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