PKS–NRPS Enzymology and Structural Biology: Considerations in Protein Production

Meredith A. Skiba; Finn P. Maloney; Qingyun Dan; Amy E. Fraley; Courtney C. Aldrich; Janet L. Smith; W. Clay Brown

doi:10.1016/bs.mie.2018.01.035

PKS–NRPS Enzymology and Structural Biology: Considerations in Protein Production

Meredith A. Skiba, Finn P. Maloney, Qingyun Dan, Amy E. Fraley, Courtney C. Aldrich, Janet L. Smith, W. Clay Brown

Medicinal Chemistry

Research output: Chapter in Book/Report/Conference proceeding › Chapter

14 Scopus citations

Abstract

The structural diversity and complexity of marine natural products have made them a rich and productive source of new bioactive molecules for drug development. The identification of these new compounds has led to extensive study of the protein constituents of the biosynthetic pathways from the producing microbes. Essential processes in the dissection of biosynthesis have been the elucidation of catalytic functions and the determination of 3D structures for enzymes of the polyketide synthases and nonribosomal peptide synthetases that carry out individual reactions. The size and complexity of these proteins present numerous difficulties in the process of going from gene to structure. Here, we review the problems that may be encountered at the various steps of this process and discuss some of the solutions devised in our and other labs for the cloning, production, purification, and structure solution of complex proteins using Escherichia coli as a heterologous host.

Original language	English (US)
Title of host publication	Methods in Enzymology
Editors	Bradley S. Moore
Publisher	Academic Press Inc.
Pages	45-88
Number of pages	44
ISBN (Print)	9780128139592
DOIs	https://doi.org/10.1016/bs.mie.2018.01.035
State	Published - Jan 1 2018

Publication series

Name	Methods in Enzymology
Volume	604
ISSN (Print)	0076-6879
ISSN (Electronic)	1557-7988

Bibliographical note

Funding Information:
This work has been supported by the NIH (R01-DK42303, R01-GM076477, R01-CA108874 to J.L.S. and R01-AI070219 to C.C.A.) and by the Margaret J. Hunter Professorship to J.L.S. M.A.S. has been supported by predoctoral fellowships from an NIH Cellular Biotechnology Training Program (T32-GM008353) and the University of Michigan Rackham Graduate School.

Publisher Copyright:
© 2018 Elsevier Inc.

Keywords

Chaperones
Coexpression
Cyanobacteria
Nonribosomal peptide synthetase
Polyketide synthase
Protein crystallization
Protein folding
Protein production
Protein purification
Protein solubility

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10.1016/bs.mie.2018.01.035

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5992914

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abstract = "The structural diversity and complexity of marine natural products have made them a rich and productive source of new bioactive molecules for drug development. The identification of these new compounds has led to extensive study of the protein constituents of the biosynthetic pathways from the producing microbes. Essential processes in the dissection of biosynthesis have been the elucidation of catalytic functions and the determination of 3D structures for enzymes of the polyketide synthases and nonribosomal peptide synthetases that carry out individual reactions. The size and complexity of these proteins present numerous difficulties in the process of going from gene to structure. Here, we review the problems that may be encountered at the various steps of this process and discuss some of the solutions devised in our and other labs for the cloning, production, purification, and structure solution of complex proteins using Escherichia coli as a heterologous host.",

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PKS–NRPS Enzymology and Structural Biology: Considerations in Protein Production

Abstract

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