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PKS-NRPS Enzymology and Structural Biology: Considerations in Protein Production.

Authors
  • Skiba, Meredith A1
  • Maloney, Finn P1
  • Dan, Qingyun1
  • Fraley, Amy E1
  • Aldrich, Courtney C2
  • Smith, Janet L3
  • Brown, W Clay4
  • 1 University of Michigan, Ann Arbor, MI, United States. , (United States)
  • 2 University of Minnesota, Minneapolis, MN, United States. , (United States)
  • 3 University of Michigan, Ann Arbor, MI, United States. Electronic address: [email protected] , (United States)
  • 4 University of Michigan, Ann Arbor, MI, United States. Electronic address: [email protected] , (United States)
Type
Published Article
Journal
Methods in enzymology
Publication Date
Jan 01, 2018
Volume
604
Pages
45–88
Identifiers
DOI: 10.1016/bs.mie.2018.01.035
PMID: 29779664
Source
Medline
Keywords
License
Unknown

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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