Examining the Link between Biofilm Formation and the Ability of Pathogenic Salmonella Strains to Colonize Multiple Host Species

MacKenzie, Keith D.; Palmer, Melissa B.; Köster, Wolfgang; White, Aaron P.

doi:10.3389/fvets.2017.00138

Cited by 83 publications

(83 citation statements)

References 185 publications

(299 reference statements)

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“…Biofilms have demonstrated roles in bacterial persistence and virulence [12,13,58,59] and have been 490 anecdotally reported in Clostridia to be composed of cellulose exopolysaccharides [7,8]. Additionally, the 491 loci encoding the molecular machinery for Pel polysaccharide has been recently identified in members of…”

mentioning

confidence: 99%

Identification of the Clostridial cellulose synthase and characterization of the cognate glycosyl hydrolase, CcsZ

Scott

Lowrance

Anderson

et al. 2019

Preprint

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1Biofilms are community structures of bacteria enmeshed in a self-produced matrix of exopolysaccharides. 2The biofilm matrix serves numerous roles, including resilience and persistence, making biofilms a subject 3 of research interest among persistent clinical pathogens of global health importance. Our current 4 understanding of the underlying biochemical pathways responsible for biosynthesis of these 5 exopolysaccharides is largely limited to Gram-negative bacteria. Clostridia are a class of Gram-positive, 6 anaerobic and spore-forming bacteria, and include the important human pathogens Clostridium 7 perfringens, Clostridium botulinum, and Clostridioides difficile, among numerous others. Clostridia have 8 been reported to form biofilms composed of cellulose, although the specific loci which encode the 9 cellulose synthase have not been identified. Here, we report the discovery of a gene cluster, which we 10 named ccsABZCD, among selected bacteria within class Clostridia that appears to encode a synthase 11 complex responsible for polymerization, modification, and export of an O-acetylated cellulose 12 exopolysaccharide. To test this hypothesis, we subcloned the putative glycoside hydrolase CcsZ and 13 solved the X-ray crystal structure of both apo-and product-bound CcsZ. Our results demonstrate that 14 CcsZ is in fact an endo-acting cellulase belonging to glycoside hydrolase family 5 . This is in 15 contrast to the Gram-negative cellulose synthase, which instead encodes BcsZ, a structurally distinct GH-16 8. We further show CcsZ is capable of hydrolysis of the soluble mock substrate carboxymethylcellulose 17 (CMC) with a pH optimum of 4.5. The data we present here serves as an entry point to an understanding 18 of biofilm formation among class Clostridia and allowed us to predict a model for Clostridial cellulose 19 synthesis.20 Author summary 21 Biofilms are communities of microorganisms that enmesh themselves in a protective matrix of elf-22 produced polysaccharide materials. Biofilms have demonstrated roles in both virulence and persistence 23 among bacterial pathogens of global health importance. The class Clostridia are a polyphyletic grouping 24 of primarily Gram-positive, anaerobic and spore-forming bacteria which contain the important and well-25 studied human pathogens Clostridioides difficile, Clostridium botulinum, and Clostridium perfringens, 26among others. Bacteria belonging to class Clostridia have been anecdotally reported to form biofilms described. In this work, we identify a gene cluster, which we name ccsABZHI, for the Clostridial cellulose 29 synthase, which bears remarkable similarity to molecular machinery required for the production of 30 cellulose biofilms in other Gram-negative bacteria. We further biochemically characterize one of these 31 enzymes, CcsZ, a predicted endoglucanase which we predicted from our model should cleave cellulose 32 exopolysaccharides. We show that CcsZ is in fact capable of this activity and belongs to a broader family 33 of glycoside hydrolases with unexpected...

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confidence: 99%

Identification of the Clostridial cellulose synthase and characterization of the cognate glycosyl hydrolase, CcsZ

Scott

Lowrance

Anderson

et al. 2019

Preprint

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“…Interestingly, representative species of the two cellulose operon lineages identified in HGT events, e.g. the plant and human pathogens, D. dadantii and S. enterica , respectively, are known to produce structurally distinct forms of cellulose with different properties and roles in pathogenesis(77,78). Furthermore, we identified that BcsB divergence was also seen to accompany the rearrangement or horizontal transfer of these operons, which further suggests that it may play a key role in the fine-tuning of cellulose production by coordinating the export of growing cellulose polymers through the periplasm.…”

Section: Discussionmentioning

confidence: 99%

A systematic pipeline for classifying bacterial operons reveals the evolutionary landscape of biofilm machineries

Bundalovic-Torma

Whitfield

Marmont

et al. 2019

Preprint

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19In bacterial functionally related genes comprising metabolic pathways and protein complexes are 20 frequently encoded in operons and are widely conserved across phylogenetically diverse species. The 21 evolution of these operon-encoded processes is affected by diverse mechanisms such gene duplication, 22 loss, rearrangement, and horizontal transfer. These mechanisms can result in functional diversification 23 of gene-families, increasing the potential evolution of novel biological pathways, and serves to adapt 24 evolutionary relationships of operon encoded genes and the evolution of operon organization as a 49 whole. To address this challenge, we present a novel method to study operon evolution by integrating 50 phylogenetic tree based clustering and genomic-context networks. We apply this approach to perform 51 the first systematic survey of all known synthase dependent bacterial biofilm machineries, 52 demonstrating the generalizability of our approach for operons of diverse size, protein family 53 composition, and species distribution. Our approach is able to identify distinct biofilm operon clades 54 across phylogenetically diverse bacteria, resulting from gene rearrangement, duplication, loss, fusion, 55 and horizontal gene transfer. We also elucidate different evolutionary trajectories of Gram-negative and 56Gram-positive biofilm production machineries, and in a companion paper (BIORXIV/2019/768473) 57 present the experimental validation of a novel mode of biofilm production in a subset of Gram-positive 58 bacteria. 59 60

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“…For example, the csgD is a transcriptional protein that activates the biosynthesis and expression of both curli and cellulose. CsgD activates transcription of the csgBAC operon, which encodes curli structural subunits, and transcription of adrA gene, involved in cellulose biosynthesis (Brombacher et al, 2006;MacKenzie et al, 2017).…”

Section: Csga Gene For Detection Of Curli Fimbria Gene (Biofilm)mentioning

confidence: 99%

Genotypic and phenotypic characteristics associated with biofilm formation in Escherichia coli and Salmonella spp. isolated from ulam in Terengganu

Bahri¹,

Abdullah²,

Lani³

et al. 2019

Food Res.

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Nowadays, the foodborne outbreaks associated with fresh produces, including ulam, are increasing worldwide. The biofilm formation or bacterial attachment to plant surface is the initial step towards the contamination in fresh produce. The biofilm phenotype of bacteria grown on congo red agar is termed as red, dry and rough (rdar) morphotype. The binding of congo red dye with both biological proteins and inert surfaces is due to the presence of curli fimbriae and cellulose as the main extracellular components. The objective of this study was to determine the rdar morphotypes, biofilm ability and the role of csgA gene of Escherichia coli and Salmonella spp. isolated from ulam or Malaysian herbs. A total of 29 isolates, including 23 E. coli and 6 Salmonella spp. were analyzed for their ability to produce biofilm by colony morphology test, microtiter plate biofilm assay and qualitative biofilm test (pellicle formation). The presence of the csgA gene of E. coli was identified by PCR, which demonstrated the potential gene that able to produce curli fimbriae. Results revealed that 16 (69.6%) E. coli isolates were categorized as strong biofilm producers, 2 (8.7%) as moderate biofilm producers, 3 (13%) as weak biofilm producers, whereas 2 (8.7%) as negative biofilm producers (did not produce biofilm). While 4 (66.7%) Salmonella spp. isolates were identified as strong biofilm producers, 1 (16.7%) as moderate biofilm producers and 1 (16.7%) as negative biofilm producers. Majority of the E. coli strains (69.6%) were identified as strong biofilm producers and able to express rdar morphotypes. The ability of the of E. coli and Salmonella spp. isolates to form biofilm reveals the ability of these isolates to persist on the fresh vegetables and become hosts for the disease transmission to humans or/and animals.

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Examining the Link between Biofilm Formation and the Ability of Pathogenic Salmonella Strains to Colonize Multiple Host Species

Cited by 83 publications

References 185 publications

Identification of the Clostridial cellulose synthase and characterization of the cognate glycosyl hydrolase, CcsZ

Identification of the Clostridial cellulose synthase and characterization of the cognate glycosyl hydrolase, CcsZ

A systematic pipeline for classifying bacterial operons reveals the evolutionary landscape of biofilm machineries

Genotypic and phenotypic characteristics associated with biofilm formation in Escherichia coli and Salmonella spp. isolated from ulam in Terengganu

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