A New Family of Capsule Polymerases Generates Teichoic Acid-Like Capsule Polymers in Gram-Negative Pathogens

ABSTRACT Group 2 capsule polymers represent crucial virulence factors of Gram-negative pathogenic bacteria. They are synthesized by enzymes called capsule polymerases. In this report, we describe a new family of polymerases that combine glycosyltransferase and hexose- and polyol-phosphate transferas...

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Autores principales:	Christa Litschko, Davide Oldrini, Insa Budde, Monika Berger, Jochen Meens, Rita Gerardy-Schahn, Francesco Berti, Mario Schubert, Timm Fiebig
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	TagF capsular polysaccharide capsule enzymatic synthesis nuclear magnetic resonance polymerases Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/6c00b5c8ad3940f2af66b90fac7376c3
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spelling	oai:doaj.org-article:6c00b5c8ad3940f2af66b90fac7376c32021-11-15T16:00:25ZA New Family of Capsule Polymerases Generates Teichoic Acid-Like Capsule Polymers in Gram-Negative Pathogens10.1128/mBio.00641-182150-7511https://doaj.org/article/6c00b5c8ad3940f2af66b90fac7376c32018-07-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00641-18https://doaj.org/toc/2150-7511ABSTRACT Group 2 capsule polymers represent crucial virulence factors of Gram-negative pathogenic bacteria. They are synthesized by enzymes called capsule polymerases. In this report, we describe a new family of polymerases that combine glycosyltransferase and hexose- and polyol-phosphate transferase activity to generate complex poly(oligosaccharide phosphate) and poly(glycosylpolyol phosphate) polymers, the latter of which display similarity to wall teichoic acid (WTA), a cell wall component of Gram-positive bacteria. Using modeling and multiple-sequence alignment, we showed homology between the predicted polymerase domains and WTA type I biosynthesis enzymes, creating a link between Gram-negative and Gram-positive cell wall biosynthesis processes. The polymerases of the new family are highly abundant and found in a variety of capsule-expressing pathogens such as Neisseria meningitidis, Actinobacillus pleuropneumoniae, Haemophilus influenzae, Bibersteinia trehalosi, and Escherichia coli with both human and animal hosts. Five representative candidates were purified, their activities were confirmed using nuclear magnetic resonance (NMR) spectroscopy, and their predicted folds were validated by site-directed mutagenesis. IMPORTANCE Bacterial capsules play an important role in the interaction between a pathogen and the immune system of its host. During the last decade, capsule polymerases have become attractive tools for the production of capsule polymers applied as antigens in glycoconjugate vaccine formulations. Conventional production of glycoconjugate vaccines requires the cultivation of the pathogen and thus the highest biosafety standards, leading to tremendous costs. With regard to animal husbandry, where vaccines could avoid the extensive use of antibiotics, conventional production is not sufficiently cost-effective. In contrast, enzymatic synthesis of capsule polymers is pathogen-free and fast, offers high stereo- and regioselectivity, and works with high efficacy. The new capsule polymerase family described here vastly increases the toolbox of enzymes available for biotechnology purposes. Representatives are abundantly found in human pathogens but also in animal pathogens, paving the way for the exploitation of polymerases for the development of a new generation of vaccines for animal husbandry.Christa LitschkoDavide OldriniInsa BuddeMonika BergerJochen MeensRita Gerardy-SchahnFrancesco BertiMario SchubertTimm FiebigAmerican Society for MicrobiologyarticleTagFcapsular polysaccharidecapsuleenzymatic synthesisnuclear magnetic resonancepolymerasesMicrobiologyQR1-502ENmBio, Vol 9, Iss 3 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	TagF capsular polysaccharide capsule enzymatic synthesis nuclear magnetic resonance polymerases Microbiology QR1-502
spellingShingle	TagF capsular polysaccharide capsule enzymatic synthesis nuclear magnetic resonance polymerases Microbiology QR1-502 Christa Litschko Davide Oldrini Insa Budde Monika Berger Jochen Meens Rita Gerardy-Schahn Francesco Berti Mario Schubert Timm Fiebig A New Family of Capsule Polymerases Generates Teichoic Acid-Like Capsule Polymers in Gram-Negative Pathogens
description	ABSTRACT Group 2 capsule polymers represent crucial virulence factors of Gram-negative pathogenic bacteria. They are synthesized by enzymes called capsule polymerases. In this report, we describe a new family of polymerases that combine glycosyltransferase and hexose- and polyol-phosphate transferase activity to generate complex poly(oligosaccharide phosphate) and poly(glycosylpolyol phosphate) polymers, the latter of which display similarity to wall teichoic acid (WTA), a cell wall component of Gram-positive bacteria. Using modeling and multiple-sequence alignment, we showed homology between the predicted polymerase domains and WTA type I biosynthesis enzymes, creating a link between Gram-negative and Gram-positive cell wall biosynthesis processes. The polymerases of the new family are highly abundant and found in a variety of capsule-expressing pathogens such as Neisseria meningitidis, Actinobacillus pleuropneumoniae, Haemophilus influenzae, Bibersteinia trehalosi, and Escherichia coli with both human and animal hosts. Five representative candidates were purified, their activities were confirmed using nuclear magnetic resonance (NMR) spectroscopy, and their predicted folds were validated by site-directed mutagenesis. IMPORTANCE Bacterial capsules play an important role in the interaction between a pathogen and the immune system of its host. During the last decade, capsule polymerases have become attractive tools for the production of capsule polymers applied as antigens in glycoconjugate vaccine formulations. Conventional production of glycoconjugate vaccines requires the cultivation of the pathogen and thus the highest biosafety standards, leading to tremendous costs. With regard to animal husbandry, where vaccines could avoid the extensive use of antibiotics, conventional production is not sufficiently cost-effective. In contrast, enzymatic synthesis of capsule polymers is pathogen-free and fast, offers high stereo- and regioselectivity, and works with high efficacy. The new capsule polymerase family described here vastly increases the toolbox of enzymes available for biotechnology purposes. Representatives are abundantly found in human pathogens but also in animal pathogens, paving the way for the exploitation of polymerases for the development of a new generation of vaccines for animal husbandry.
format	article
author	Christa Litschko Davide Oldrini Insa Budde Monika Berger Jochen Meens Rita Gerardy-Schahn Francesco Berti Mario Schubert Timm Fiebig
author_facet	Christa Litschko Davide Oldrini Insa Budde Monika Berger Jochen Meens Rita Gerardy-Schahn Francesco Berti Mario Schubert Timm Fiebig
author_sort	Christa Litschko
title	A New Family of Capsule Polymerases Generates Teichoic Acid-Like Capsule Polymers in Gram-Negative Pathogens
title_short	A New Family of Capsule Polymerases Generates Teichoic Acid-Like Capsule Polymers in Gram-Negative Pathogens
title_full	A New Family of Capsule Polymerases Generates Teichoic Acid-Like Capsule Polymers in Gram-Negative Pathogens
title_fullStr	A New Family of Capsule Polymerases Generates Teichoic Acid-Like Capsule Polymers in Gram-Negative Pathogens
title_full_unstemmed	A New Family of Capsule Polymerases Generates Teichoic Acid-Like Capsule Polymers in Gram-Negative Pathogens
title_sort	new family of capsule polymerases generates teichoic acid-like capsule polymers in gram-negative pathogens
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/6c00b5c8ad3940f2af66b90fac7376c3
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A New Family of Capsule Polymerases Generates Teichoic Acid-Like Capsule Polymers in Gram-Negative Pathogens

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