Bacterial Polysaccharides

Methods and Protocols

Buch | Hardcover

312 Seiten

2019 | 1st ed. 2019
Humana Press Inc. (Verlag)
978-1-4939-9153-2 (ISBN)

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This book provides a selection of recently developed methods and protocols in bacterial glycomics to aid in bettering our understanding of the structures and functions of bacterial polysaccharides, their attachments to proteins and lipids, their role in biofilm formation, as well as their biosynthesis. With the emerging bacterial resistance to commonly used antibiotics world-wide, these techniques to study the outer polysaccharides of bacteria, with their functions in bacterial adhesion, colonization, growth, establishment of biofilms, and control virulence and pathogenicity, are increasingly important. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Bacterial Polysaccharides: Methods and Protocols aims to support researchers contributing to future approaches that will fill our knowledge gaps and define anti-bacterial targets.

1. Customisable cloning of whole polysaccharide gene clusters by yeast homologous recombination.- 2. RecET Direct Cloning of Polysaccharide Gene Cluster from Gram-negative Bacteria.- 3. Purification of Capsular Polysaccharide Complex from Gram Negative Bacteria.- 4. Simple protocol to purify cell wall polysaccharide from Gram+ bacteria and assess its structural integrity.- 5. Extraction and purification of wall bound polymers of Gram-positive bacteria.- 6. Purification and Analysis of Mycobacterial Phosphatidylinositol Mannosides, Lipomannan and Lipoarabinomannan.- 7. Conjugation of synthetic oligosaccharides to proteins by squaric acid chemistry.- 8. Direct conjugation of bacterial polysaccharides to proteins by squaric acid chemistry.- 9. Chemical Modification of the N-Acetyl Moieties of Hyaluronic Acid from Streptococcus equi for studies in cytokine production.- 10. O-Acetylation of Bacterial Cell Wall Polysaccharides.- 11. Designing glycosyltransferase expression constructs for improved purification, protein yield and crystallization.- 12. Utilization of fluorescently-tagged synthetic acceptor molecules for in vitro characterization of a dual-domain glycosyltransferase enzyme, KpsC, from Escherichia coli.- 13. Synthesis of Phenoxyundecyl Diphosphate Disaccharides for Studies of the Biosynthesis of O-Antigenic Polysaccharides in Enteric Bacteria.- 14. Characterization of arabinosyl transfer reactions in the biosynthesis of mycobacterial cell envelope (lipo)polysaccharides.- 15. Enzymatic synthesis of the repeating unit oligosaccharides of Escherichia coli O104.- 16. Sulfhydryl labelling as a tool to investigate the topology of membrane proteins involved in lipopolysaccharide biosynthesis.- 17. Assay Methods for the Glycosyltransferases Involved in Synthesis of Bacterial Polysaccharides.- 18. Glycosyltransferase Activity Assays Using Colorimetric Methods.- 19. In vitro characterization of a multidomain glycosyltransferase using fluorescently-tagged synthetic acceptors.- 20. Glycosyltransferase-coupled assays for 4-epimerase WbpP from Pseudomonas aeruginosa.- 21. Assaying fucosidase activity.- 22. In vitro synthesis of bioactive glycovariants of enterocin 96, an antimicrobial peptide from Enterococcus faecalis.- 23. High-Throughput Screening for Inhibitors of Wall Teichoic Acid Biosynthesis in Staphylococcus aureus.

Erscheinungsdatum	29.03.2019
Reihe/Serie	Methods in Molecular Biology ; 1954
Zusatzinfo	41 Illustrations, color; 29 Illustrations, black and white; XIII, 312 p. 70 illus., 41 illus. in color.
Verlagsort	Totowa, NJ
Sprache	englisch
Maße	178 x 254 mm
Themenwelt	Studium ► Querschnittsbereiche ► Infektiologie / Immunologie
	Naturwissenschaften ► Biologie ► Biochemie
	Naturwissenschaften ► Biologie ► Mikrobiologie / Immunologie
Schlagworte	antibacterials • Antibiotic resistantance • biosynthesis • Biosynthetic enzymes • glycoconjugates • Glycohydrolases • glycosyltransferases • Vaccine development
ISBN-10	1-4939-9153-1 / 1493991531
ISBN-13	978-1-4939-9153-2 / 9781493991532
Zustand	Neuware