Objective: Most protein secretion systems are found in gram-negative bacteria, but the mechanism of endoglucanase (BcsZ) secretion in Escherichia coli (E. coli) remains unclear.
Methods: In this study, we used JBZ-DH5α (which overexpresses BcsZ on the E. coli DH5α genome) as the initial strain. A mutant library was created by randomly inserting the TN5 transposon into the genome, and mutants with reduced transparent circles were identified on Congo red plates. The insertion sites of transposons in the genome were determined through whole-genome sequencing.
Results: The results revealed that the genes rnc, lon, and suhB, which encode RNC-ribonuclease III (RNC), LON-protease (LON), and SuhB-inositol phosphatase (SuhB), respectively, were disrupted. BcsZ secretion decreased in E. coli DH5α when the lon, rnc, or suhB genes were deleted, but the overexpression of these genes restored their secretion levels.
Conclusion: These findings suggest that the lon, rnc, and suhB genes play a role in BcsZ secretion in E. coli, potentially enhancing our knowledge of BcsZ secretion and offering a strategy to increase protein secretion in E. coli as a cell factory.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.gene.2024.148936 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Impact of a random TN5 mutation on endoglucanase secretion in ruminal cellulolytic Escherichia coli.
Gene
January 2025
Center for Energy Conservation and Emission Reduction in Fermentation Industry in Inner Mongolia, Hohhot 010051, Inner Mongolia, China; Engineering Research Center of Inner Mongolia for Green Manufacturing in Bio-fermentation Industry, Hohhot 010051, Inner Mongolia, China; Specialized Technology Research and Pilot Public Service Platform for Biological Fermentation in Inner Mongolia, Hohhot 010051, Inner Mongolia, China; College of Chemical Engineering, Inner Mongolia University of Technology, Hohhot 010051, Inner Mongolia, China. Electronic address:
Objective: Most protein secretion systems are found in gram-negative bacteria, but the mechanism of endoglucanase (BcsZ) secretion in Escherichia coli (E. coli) remains unclear.
Methods: In this study, we used JBZ-DH5α (which overexpresses BcsZ on the E.
Insights into phosphoethanolamine cellulose synthesis and secretion across the Gram-negative cell envelope.
Nat Commun
September 2024
Department of Molecular Physiology and Biological Physics, University of Virginia School of Medicine, Charlottesville, VA, USA.
Phosphoethanolamine (pEtN) cellulose is a naturally occurring modified cellulose produced by several Enterobacteriaceae. The minimal components of the E. coli cellulose synthase complex include the catalytically active BcsA enzyme, a hexameric semicircle of the periplasmic BcsB protein, and the outer membrane (OM)-integrated BcsC subunit containing periplasmic tetratricopeptide repeats (TPR).
View Article and Find Full Text PDFMolecular insights into phosphoethanolamine cellulose formation and secretion.
bioRxiv
April 2024
Department of Molecular Physiology and Biological Physics, University of Virginia School of Medicine, Charlottesville, VA 22903, USA.
Phosphoethanolamine (pEtN) cellulose is a naturally occurring modified cellulose produced by several Enterobacteriaceae. The minimal components of the cellulose synthase complex include the catalytically active BcsA enzyme, an associated periplasmic semicircle of hexameric BcsB, as well as the outer membrane (OM)-integrated BcsC subunit containing periplasmic tetratricopeptide repeats (TPR). Additional subunits include BcsG, a membrane-anchored periplasmic pEtN transferase associated with BcsA, and BcsZ, a conserved periplasmic cellulase of unknown biological function.
View Article and Find Full Text PDFBacteriophage S6 requires bacterial cellulose for Erwinia amylovora infection.
Environ Microbiol
August 2022
Food Microbiology Research Group, Institute of Food and Beverage Innovation, Zurich University of Applied Sciences (ZHAW), Wädenswil, Switzerland.
Bacteriophages are highly selective in targeting bacteria. This selectivity relies on the specific adsorption of phages to the host cell surface. In this study, a Tn5 transposon mutant library of Erwinia amylovora, the causative agent of fire blight, was screened to identify bacterial receptors required for infection by the podovirus S6.
View Article and Find Full Text PDFReverse vaccinology approach for the identifications of potential vaccine candidates against Salmonella.
Int J Med Microbiol
July 2021
School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai, PR China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, PR China. Electronic address:
Salmonella is a leading cause of foodborne pathogen which causes intestinal and systemic diseases across the world. Vaccination is the most effective protection against Salmonella, but the identification and design of an effective broad-spectrum vaccine is still a great challenge, because of the multi-serotypes of Salmonella. Reverse vaccinology is a new tool to discovery and design vaccine antigens combining human immunology, structural biology and computational biology with microbial genomics.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!