Pellicle formation by Escherichia coli K-12: role of adhesins and motility

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Pellicle formation by Escherichia coli K-12 : role of adhesins and motility. / Golub, Stacey; Overton, Tim.

In: Journal of Bioscience and Bioengineering, Vol. 131, No. 4, 04.2021, p. 381-389.

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@article{ef407a090f3c4416be39d7fd4f1f3a96,
title = "Pellicle formation by Escherichia coli K-12: role of adhesins and motility",
abstract = "Initial work to generate physically robust biofilms for biocatalytic applications revealed that Escherichia coli K-12 can form a floating biofilm at the air-liquid interface, commonly referred to as a pellicle. Unlike other species where pellicle formation is well-characterised, such as Bacillus subtilis, there are few reports of E. coli K-12 pellicles in the literature. In order to study pellicle formation, a growth model was developed and pellicle formation was monitored over time. Mechanical forces, both motility and shaking, were shown to have effects on pellicle formation and development. The role and regulation of curli, an amyloid protein adhesin critical in E. coli K-12 biofilm formation, was studied by using promoter-green fluorescent protein reporters; flow cytometry and confocal laser scanning microscopy were used to monitor curli expression over time and in different locations. Curli were found to be not only crucial for pellicle formation, but also heterogeneously expressed within the pellicle. The components of the extracellular polymeric substances (EPS) in pellicles were analysed by confocal microscopy using lectins, revealing distinct pellicle morphology on the air-facing and medium-facing sides, and spatially- and temporally-regulated generation of the EPS components poly-N-acetyl glucosamine and colanic acid. We discuss the difference between pellicles formed by E. coli K-12, pathogenic E. coli strains and other species, and the relationship between E. coli K-12 pellicles and solid surface-attached biofilms. ",
keywords = "Colanic acid, Confocal microscopy, Curli, Motility, Poly-N-acetyl glucosamine",
author = "Stacey Golub and Tim Overton",
note = "Acknowledgments: Genome sequencing was provided by MicrobesNG, which is supported by the UK Biotechnology and Biological Sciences Research Council (BBSRC grant number BB/L024209/1). We thank Paolo Landini for pT7-CsgD and pT7-7 and James Leech for pJLC-T. We are extremely grateful to Alessandro Di Maio at the Birmingham Advanced Light Microscopy facility for confocal microscopy assistance and Charles Penn for discussions regarding hypermotility.",
year = "2021",
month = apr,
doi = "10.1016/j.jbiosc.2020.12.002",
language = "English",
volume = "131",
pages = "381--389",
journal = "Journal of Bioscience and Bioengineering",
issn = "1389-1723",
publisher = "Elsevier",
number = "4",

}

RIS

TY - JOUR

T1 - Pellicle formation by Escherichia coli K-12

T2 - role of adhesins and motility

AU - Golub, Stacey

AU - Overton, Tim

N1 - Acknowledgments: Genome sequencing was provided by MicrobesNG, which is supported by the UK Biotechnology and Biological Sciences Research Council (BBSRC grant number BB/L024209/1). We thank Paolo Landini for pT7-CsgD and pT7-7 and James Leech for pJLC-T. We are extremely grateful to Alessandro Di Maio at the Birmingham Advanced Light Microscopy facility for confocal microscopy assistance and Charles Penn for discussions regarding hypermotility.

PY - 2021/4

Y1 - 2021/4

N2 - Initial work to generate physically robust biofilms for biocatalytic applications revealed that Escherichia coli K-12 can form a floating biofilm at the air-liquid interface, commonly referred to as a pellicle. Unlike other species where pellicle formation is well-characterised, such as Bacillus subtilis, there are few reports of E. coli K-12 pellicles in the literature. In order to study pellicle formation, a growth model was developed and pellicle formation was monitored over time. Mechanical forces, both motility and shaking, were shown to have effects on pellicle formation and development. The role and regulation of curli, an amyloid protein adhesin critical in E. coli K-12 biofilm formation, was studied by using promoter-green fluorescent protein reporters; flow cytometry and confocal laser scanning microscopy were used to monitor curli expression over time and in different locations. Curli were found to be not only crucial for pellicle formation, but also heterogeneously expressed within the pellicle. The components of the extracellular polymeric substances (EPS) in pellicles were analysed by confocal microscopy using lectins, revealing distinct pellicle morphology on the air-facing and medium-facing sides, and spatially- and temporally-regulated generation of the EPS components poly-N-acetyl glucosamine and colanic acid. We discuss the difference between pellicles formed by E. coli K-12, pathogenic E. coli strains and other species, and the relationship between E. coli K-12 pellicles and solid surface-attached biofilms.

AB - Initial work to generate physically robust biofilms for biocatalytic applications revealed that Escherichia coli K-12 can form a floating biofilm at the air-liquid interface, commonly referred to as a pellicle. Unlike other species where pellicle formation is well-characterised, such as Bacillus subtilis, there are few reports of E. coli K-12 pellicles in the literature. In order to study pellicle formation, a growth model was developed and pellicle formation was monitored over time. Mechanical forces, both motility and shaking, were shown to have effects on pellicle formation and development. The role and regulation of curli, an amyloid protein adhesin critical in E. coli K-12 biofilm formation, was studied by using promoter-green fluorescent protein reporters; flow cytometry and confocal laser scanning microscopy were used to monitor curli expression over time and in different locations. Curli were found to be not only crucial for pellicle formation, but also heterogeneously expressed within the pellicle. The components of the extracellular polymeric substances (EPS) in pellicles were analysed by confocal microscopy using lectins, revealing distinct pellicle morphology on the air-facing and medium-facing sides, and spatially- and temporally-regulated generation of the EPS components poly-N-acetyl glucosamine and colanic acid. We discuss the difference between pellicles formed by E. coli K-12, pathogenic E. coli strains and other species, and the relationship between E. coli K-12 pellicles and solid surface-attached biofilms.

KW - Colanic acid

KW - Confocal microscopy

KW - Curli

KW - Motility

KW - Poly-N-acetyl glucosamine

UR - http://www.scopus.com/inward/record.url?scp=85099716076&partnerID=8YFLogxK

U2 - 10.1016/j.jbiosc.2020.12.002

DO - 10.1016/j.jbiosc.2020.12.002

M3 - Article

VL - 131

SP - 381

EP - 389

JO - Journal of Bioscience and Bioengineering

JF - Journal of Bioscience and Bioengineering

SN - 1389-1723

IS - 4

ER -