Colocalization and Disposition of Cellulosomes in <italic toggle="yes">Clostridium clariflavum</italic> as Revealed by Correlative Superresolution Imaging

ABSTRACT Cellulosomes are multienzyme complexes produced by anaerobic, cellulolytic bacteria for highly efficient breakdown of plant cell wall polysaccharides. Clostridium clariflavum is an anaerobic, thermophilic bacterium that produces the largest assembled cellulosome complex in nature to date, c...

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Autores principales:	Lior Artzi, Tali Dadosh, Elad Milrot, Sarah Moraïs, Smadar Levin-Zaidman, Ely Morag, Edward A. Bayer
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	CLEM SEM STORM bacterial cell surface fluorescence microscopy protuberance-like structures Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/06162ed2e7ca411a91dac5586024e352
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spelling	oai:doaj.org-article:06162ed2e7ca411a91dac5586024e3522021-11-15T15:53:25ZColocalization and Disposition of Cellulosomes in <italic toggle="yes">Clostridium clariflavum</italic> as Revealed by Correlative Superresolution Imaging10.1128/mBio.00012-182150-7511https://doaj.org/article/06162ed2e7ca411a91dac5586024e3522018-03-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00012-18https://doaj.org/toc/2150-7511ABSTRACT Cellulosomes are multienzyme complexes produced by anaerobic, cellulolytic bacteria for highly efficient breakdown of plant cell wall polysaccharides. Clostridium clariflavum is an anaerobic, thermophilic bacterium that produces the largest assembled cellulosome complex in nature to date, comprising three types of scaffoldins: a primary scaffoldin, ScaA; an adaptor scaffoldin, ScaB; and a cell surface anchoring scaffoldin, ScaC. This complex can contain 160 polysaccharide-degrading enzymes. In previous studies, we proposed potential types of cellulosome assemblies in C. clariflavum and demonstrated that these complexes are released into the extracellular medium. In the present study, we explored the disposition of the highly structured, four-tiered cell-anchored cellulosome complex of this bacterium. Four separate, integral cellulosome components were subjected to immunolabeling: ScaA, ScaB, ScaC, and the cellulosome’s most prominent enzyme, GH48. Imaging of the cells by correlating scanning electron microscopy and three-dimensional (3D) superresolution fluorescence microscopy revealed that some of the protuberance-like structures on the cell surface represent cellulosomes and that the components are highly colocalized and organized by a defined hierarchy on the cell surface. The display of the cellulosome on the cell surface was found to differ between cells grown on soluble or insoluble substrates. Cell growth on microcrystalline cellulose and wheat straw exhibited dramatic enhancement in the amount of cellulosomes displayed on the bacterial cell surface. IMPORTANCE Conversion of plant biomass into soluble sugars is of high interest for production of fermentable industrial materials, such as biofuels. Biofuels are a very attractive alternative to fossil fuels, both for recycling of agricultural wastes and as a source of sustainable energy. Cellulosomes are among the most efficient enzymatic degraders of biomass known to date, due to the incorporation of a multiplicity of enzymes into a potent, multifunctional nanomachine. The intimate association with the bacterial cell surface is inherent in its efficient action on lignocellulosic substrates, although this property has not been properly addressed experimentally. The dramatic increase in cellulosome performance on recalcitrant feedstocks is critical for the design of cost-effective processes for efficient biomass degradation.Lior ArtziTali DadoshElad MilrotSarah MoraïsSmadar Levin-ZaidmanEly MoragEdward A. BayerAmerican Society for MicrobiologyarticleCLEMSEMSTORMbacterial cell surfacefluorescence microscopyprotuberance-like structuresMicrobiologyQR1-502ENmBio, Vol 9, Iss 1 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	CLEM SEM STORM bacterial cell surface fluorescence microscopy protuberance-like structures Microbiology QR1-502
spellingShingle	CLEM SEM STORM bacterial cell surface fluorescence microscopy protuberance-like structures Microbiology QR1-502 Lior Artzi Tali Dadosh Elad Milrot Sarah Moraïs Smadar Levin-Zaidman Ely Morag Edward A. Bayer Colocalization and Disposition of Cellulosomes in <italic toggle="yes">Clostridium clariflavum</italic> as Revealed by Correlative Superresolution Imaging
description	ABSTRACT Cellulosomes are multienzyme complexes produced by anaerobic, cellulolytic bacteria for highly efficient breakdown of plant cell wall polysaccharides. Clostridium clariflavum is an anaerobic, thermophilic bacterium that produces the largest assembled cellulosome complex in nature to date, comprising three types of scaffoldins: a primary scaffoldin, ScaA; an adaptor scaffoldin, ScaB; and a cell surface anchoring scaffoldin, ScaC. This complex can contain 160 polysaccharide-degrading enzymes. In previous studies, we proposed potential types of cellulosome assemblies in C. clariflavum and demonstrated that these complexes are released into the extracellular medium. In the present study, we explored the disposition of the highly structured, four-tiered cell-anchored cellulosome complex of this bacterium. Four separate, integral cellulosome components were subjected to immunolabeling: ScaA, ScaB, ScaC, and the cellulosome’s most prominent enzyme, GH48. Imaging of the cells by correlating scanning electron microscopy and three-dimensional (3D) superresolution fluorescence microscopy revealed that some of the protuberance-like structures on the cell surface represent cellulosomes and that the components are highly colocalized and organized by a defined hierarchy on the cell surface. The display of the cellulosome on the cell surface was found to differ between cells grown on soluble or insoluble substrates. Cell growth on microcrystalline cellulose and wheat straw exhibited dramatic enhancement in the amount of cellulosomes displayed on the bacterial cell surface. IMPORTANCE Conversion of plant biomass into soluble sugars is of high interest for production of fermentable industrial materials, such as biofuels. Biofuels are a very attractive alternative to fossil fuels, both for recycling of agricultural wastes and as a source of sustainable energy. Cellulosomes are among the most efficient enzymatic degraders of biomass known to date, due to the incorporation of a multiplicity of enzymes into a potent, multifunctional nanomachine. The intimate association with the bacterial cell surface is inherent in its efficient action on lignocellulosic substrates, although this property has not been properly addressed experimentally. The dramatic increase in cellulosome performance on recalcitrant feedstocks is critical for the design of cost-effective processes for efficient biomass degradation.
format	article
author	Lior Artzi Tali Dadosh Elad Milrot Sarah Moraïs Smadar Levin-Zaidman Ely Morag Edward A. Bayer
author_facet	Lior Artzi Tali Dadosh Elad Milrot Sarah Moraïs Smadar Levin-Zaidman Ely Morag Edward A. Bayer
author_sort	Lior Artzi
title	Colocalization and Disposition of Cellulosomes in <italic toggle="yes">Clostridium clariflavum</italic> as Revealed by Correlative Superresolution Imaging
title_short	Colocalization and Disposition of Cellulosomes in <italic toggle="yes">Clostridium clariflavum</italic> as Revealed by Correlative Superresolution Imaging
title_full	Colocalization and Disposition of Cellulosomes in <italic toggle="yes">Clostridium clariflavum</italic> as Revealed by Correlative Superresolution Imaging
title_fullStr	Colocalization and Disposition of Cellulosomes in <italic toggle="yes">Clostridium clariflavum</italic> as Revealed by Correlative Superresolution Imaging
title_full_unstemmed	Colocalization and Disposition of Cellulosomes in <italic toggle="yes">Clostridium clariflavum</italic> as Revealed by Correlative Superresolution Imaging
title_sort	colocalization and disposition of cellulosomes in <italic toggle="yes">clostridium clariflavum</italic> as revealed by correlative superresolution imaging
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/06162ed2e7ca411a91dac5586024e352
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Colocalization and Disposition of Cellulosomes in <italic toggle="yes">Clostridium clariflavum</italic> as Revealed by Correlative Superresolution Imaging

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