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.|
|Number of pages||44|
|State||Published - Jan 1 2018|
|Name||Methods in Enzymology|
Bibliographical noteFunding 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.
© 2018 Elsevier Inc.
- Nonribosomal peptide synthetase
- Polyketide synthase
- Protein crystallization
- Protein folding
- Protein production
- Protein purification
- Protein solubility