Utilizing the ABC Transporter for Growth Factor Production by fleQ Deletion Mutant of Pseudomonas fluorescens

Pseudomonas fluorescens, a gram-negative bacterium, has been proven to be a capable protein manufacturing factory (PMF). Utilizing its ATP-binding cassette (ABC) transporter, a type I secretion system, P. fluorescens has successfully produced recombinant proteins. However, besides the target protein...

Full description

Saved in:
Bibliographic Details
Published in:Biomedicines 2021-06, Vol.9 (6), p.679
Main Authors: Fabia, Benedict-Uy, Bingwa, Joshua, Park, Jiyeon, Hieu, Nguyen-Mihn, Ahn, Jung-Hoon
Format: Article
Language:English
Subjects:
ABC transporter
Deletion mutant
E coli
Fibroblast growth factor 1
Flagella
Flagellin
fleQ
Gene deletion
Gene expression
Genes
Genomes
growth factor
Insulin
Insulin-like growth factors
Motility
Mutants
Mutation
negatively supercharged protein
Nerve growth factor
Peptides
Plasmids
Protein purification
Proteins
Pseudomonas fluorescens
Tumor necrosis factor-β
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Pseudomonas fluorescens, a gram-negative bacterium, has been proven to be a capable protein manufacturing factory (PMF). Utilizing its ATP-binding cassette (ABC) transporter, a type I secretion system, P. fluorescens has successfully produced recombinant proteins. However, besides the target proteins, P. fluorescens also secretes unnecessary background proteins that complicate protein purification and other downstream processes. One of the background proteins produced in large amounts is FliC, a flagellin protein. In this study, the master regulator of flagella gene expression, fleQ, was deleted from P. fluorescens Δtp, a lipase and protease double-deletion mutant, via targeted gene knockout. FleQ directs flagella synthesis, so the new strain, P. fluorescens ΔfleQ, does not produce flagella-related proteins. This not only simplifies purification but also makes P. fluorescens ΔfleQ an eco-friendly expression host because it will not survive outside a controlled environment. Six recombinant growth factors, namely, insulin-like growth factors I and II, beta-nerve growth factor, fibroblast growth factor 1, transforming growth factor beta, and tumor necrosis factor beta, prepared using our supercharging method, were successfully secreted by P. fluorescens ΔfleQ. Our findings demonstrate the potential of P. fluorescens ΔfleQ, combined with our supercharging process, as a PMF.
ISSN:2227-9059
2227-9059
DOI:10.3390/biomedicines9060679