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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F20%3A00114419" target="_blank" >RIV/00216224:14310/20:00114419 - isvavai.cz</a>

Výsledek na webu

<a href="https://doi.org/10.1021/acssynbio.0c00276" target="_blank" >https://doi.org/10.1021/acssynbio.0c00276</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1021/acssynbio.0c00276" target="_blank" >10.1021/acssynbio.0c00276</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

<a href="/cs/project/GJ19-06511Y" target="_blank" >GJ19-06511Y: Ortogonalizace metabolismu sacharidů v bakteriálním šasi Pseudomonas putida EM42 pro ko-utilizaci cukrů z rostlinné biomasy</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2020

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

ACS Synthetic Biology

ISSN

2161-5063

e-ISSN

—

Svazek periodika

9

Číslo periodika v rámci svazku

10

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

16

Strana od-do

2749-2764

Kód UT WoS článku

000582582100014

EID výsledku v databázi Scopus

2-s2.0-85093538854