Regulating exopolysaccharide gene wcaF allows control of Escherichia coli biofilm formation

Zhang, Jingyun; Poh, Chueh Loo

doi:10.1038/s41598-018-31161-7

Cited by 24 publications

(13 citation statements)

References 48 publications

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“…However, we cannot rule out the possibility that a second acetylation takes place after formation of the oligosaccharide or polymer. The importance of WcaF is consistent with studies showing that this protein is critical to the formation of colanic acid in vivo and, more recently, the importance of that gene for biofilm formation. , We have also shown that WcaB, the other encoded acetyltransferase, had no influence on the disaccharide but enhances the ability of WcaA to add the fifth sugar to the CA repeating unit and also appears to be a critical protein in the assembly of the oligosaccharide repeat unit.…”

Section: Discussionsupporting

confidence: 89%

Identification of the Functional Roles of Six Key Proteins in the Biosynthesis of Enterobacteriaceae Colanic Acid

2019

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When introduced to harsh conditions such as low pH, pathogenic Escherichia coli can secrete colanic acid to establish a protective barrier between the organism and the acidic environment. The colanic acid is made up of a six-sugar repeating unit polymer comprised of glucose, fucose, galactose, and glucuronic acid. The region of the E. coli genome that encodes for colanic acid biosynthesis has been reported, and the first enzyme in the biosynthesis pathway has been biochemically characterized. However, the specific roles of the remaining genes required for colanic acid biosynthesis have not been identified. Here we report the in vitro reconstitution of the next six steps in the assembly of the colanic acid repeating unit. To do this, we have cloned and overexpressed each gene within the colanic acid biosynthesis operon. We then tested the activity of the protein product of these genes using high performance liquid chromatography (HPLC) analysis and a fluorescent analogue of the isoprenoid anchor bactoprenyl diphospho-glucose as a starting substrate. To ensure that retention time changes were associated with varying sugar additions or modifications, we developed a liquid chromatography mass spectrometry (LC-MS) method for analysis of the products produced by each enzyme. We have identified the function of all but one encoded glycosyltransferase and have identified the function of two acetyltransferases. This work demonstrates the centrality of acetylation in the biosynthesis of colanic acid and provides insight into the activity of key proteins involved in the production of an important and highly conserved bacterial glycopolymer.

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Section: Discussionsupporting

confidence: 89%

Identification of the Functional Roles of Six Key Proteins in the Biosynthesis of Enterobacteriaceae Colanic Acid

2019

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“…Biofilm thickness was detected according to a previously published method, with minor modification [ 11 ]. After incubation, the microscope slides with biofilm from each group were gently rinsed with deionised water to remove unattached cells and then stained with SYTO9/propidium iodide according to the instructions of the L13152 LIVE/DEAD BacLight bacterial viability kit (Invitrogen Molecular Probes, USA).…”

Section: Methodsmentioning

confidence: 99%

Label-free quantitative proteomic analysis of the inhibition effect of Lactobacillus rhamnosus GG on Escherichia coli biofilm formation in co-culture

Song

Lou

et al. 2021

Proteome Sci

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Background Escherichia coli (E. coli) is the principal pathogen that causes biofilm formation. Biofilms are associated with infectious diseases and antibiotic resistance. This study employed proteomic analysis to identify differentially expressed proteins after coculture of E. coli with Lactobacillus rhamnosus GG (LGG) microcapsules. Methods To explore the relevant protein abundance changes after E. coli and LGG coculture, label-free quantitative proteomic analysis and qRT-PCR were applied to E. coli and LGG microcapsule groups before and after coculture, respectively. Results The proteomic analysis characterised a total of 1655 proteins in E. coli K12MG1655 and 1431 proteins in the LGG. After coculture treatment, there were 262 differentially expressed proteins in E. coli and 291 in LGG. Gene ontology analysis showed that the differentially expressed proteins were mainly related to cellular metabolism, the stress response, transcription and the cell membrane. A protein interaction network and Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway analysis indicated that the differentiated proteins were mainly involved in the protein ubiquitination pathway and mitochondrial dysfunction. Conclusions These findings indicated that LGG microcapsules may inhibit E. coli biofilm formation by disrupting metabolic processes, particularly in relation to energy metabolism and stimulus responses, both of which are critical for the growth of LGG. Together, these findings increase our understanding of the interactions between bacteria under coculture conditions.

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“…CA production is not required for surface attachment, however, CA is critical for the formation of the complex three-dimensional structure and depth of E. coli biofilms [ 63 ]. Bacterial biofilms have been described as sessile bacterial communities that live attached to each other and stick onto surfaces [ 68 , 69 ]. Biofilm is so abundant in natural environments, but can also cause persistent, antibiotic-resistant infections [ 68 , 70 ], especially impinge significantly upon our industrialized world [ 71 , 72 ].…”

Section: Polysaccharides Of Lps Consume Lots Of Nutrients and Influenmentioning

confidence: 99%

Insights into the structure of Escherichia coli outer membrane as the target for engineering microbial cell factories

Wang

2021

Microb Cell Fact

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Escherichia coli is generally used as model bacteria to define microbial cell factories for many products and to investigate regulation mechanisms. E. coli exhibits phospholipids, lipopolysaccharides, colanic acid, flagella and type I fimbriae on the outer membrane which is a self-protective barrier and closely related to cellular morphology, growth, phenotypes and stress adaptation. However, these outer membrane associated molecules could also lead to potential contamination and insecurity for fermentation products and consume lots of nutrients and energy sources. Therefore, understanding critical insights of these membrane associated molecules is necessary for building better microbial producers. Here the biosynthesis, function, influences, and current membrane engineering applications of these outer membrane associated molecules were reviewed from the perspective of synthetic biology, and the potential and effective engineering strategies on the outer membrane to improve fermentation features for microbial cell factories were suggested.

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Regulating exopolysaccharide gene wcaF allows control of Escherichia coli biofilm formation

Cited by 24 publications

References 48 publications

Identification of the Functional Roles of Six Key Proteins in the Biosynthesis of Enterobacteriaceae Colanic Acid

Identification of the Functional Roles of Six Key Proteins in the Biosynthesis of Enterobacteriaceae Colanic Acid

Label-free quantitative proteomic analysis of the inhibition effect of Lactobacillus rhamnosus GG on Escherichia coli biofilm formation in co-culture

Insights into the structure of Escherichia coli outer membrane as the target for engineering microbial cell factories

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