By Bradley Evans
This quantity offers an summary of nonribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) constitution and serve as. It then maintains with tools for the research of those pathways together with traditional enzymological assays, modern mass spectrometric research strategies, really expert molecular organic ways acceptable to NRPSs and PKSs, and small molecule research instruments adapted to this very distinctive category of ordinary items, and concludes by way of analyzing bioinformatics instruments for the research of those enzymes, pathways, and molecules. Written for the hugely winning Methods in Molecular Biology sequence, chapters comprise introductions to their respective themes, lists of the mandatory fabrics and reagents, step by step, without problems reproducible laboratory protocols, and tips about troubleshooting and warding off identified pitfalls.
Authoritative and useful, Nonribosomal Peptide and Polyketide Biosynthesis: tools and Protocols serves as a invaluable reference for these skilled in learning NRPS and PKS enzymes, pathways, and usual items in addition to a gateway for these simply coming into the field.
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Extra resources for Nonribosomal Peptide and Polyketide Biosynthesis: Methods and Protocols
Sample text
Proc Natl Acad Sci U S A 99:12120–12125 21. Hamoen LW, Eshuis H, Jongbloed J et al (1995) A small gene, designated comS, located within the coding region of the fourth amino acid-activation domain of srfA, is required for competence development in Bacillus subtilis. Mol Microbiol 15:55–63 22. Reger AS, Carney JM, Gulick AM (2007) Biochemical and crystallographic analysis of substrate binding and conformational changes in acetyl-CoA synthetase. Biochemistry 46:6536–6546 28 Bradley R. Miller and Andrew M.
Type III PKSs, which are widely distributed amongst bacteria, fungi, and plants, are in essence free standing KS domains that act to catalyze the sequential condensation of acetate units to a CoA derived starter unit [33–35]. Chemical diversity in the products of type III systems arises due to the choice of starter unit, the number of chain extension steps catalyzed, the mechanism of cyclization, and product modification through the action of allied tailoring enzymes. 3 The DEBS Paradigm Early attempts to delineate the enzymology of modular PKSs focused on the deoxyerythronolide B synthase (DEBS).
In extreme cases their proteinogenic mass can exceed 5 MDa. The distinguishing structural feature of these systems is their assembly line-like architecture, which comprises a series of linearly arranged, multi-domain extension modules, Assembly Line Enzymology of PKSs 33 Fig. 2 (a) Chemical structures of example PKS starter and extender units. (b) Individual steps involved in extender unit selection and chain incorporation. (c) Illustrative chain modifications catalyzed by module embedded PKS domains housed in sequence within giant polypeptide chains [1, 16].