PKS-NRPS Enzymology and Structural Biology: Considerations in Protein Production.
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.
['Skiba|Meredith A|MA|', 'Maloney|Finn P|FP|', 'Dan|Qingyun|Q|', 'Fraley|Amy E|AE|', 'Aldrich|Courtney C|CC|', 'Smith|Janet L|JL|', 'Brown|W Clay|WC|']
D000213:Acyl Carrier Protein D001419:Bacteria D001425:Bacterial Outer Membrane Proteins D003001:Cloning, Molecular D003062:Codon D003460:Crystallization D003470:Culture Media D004926:Escherichia coli D029968:Escherichia coli Proteins D010453:Peptide Synthases D010957:Plasmids D048630:Polyketide Synthases D011401:Promoter Regions, Genetic D000072417:Protein Domains D015202:Protein Engineering D011994:Recombinant Proteins
Protein production Polyketide synthase Nonribosomal peptide synthetase Chaperones Coexpression Cyanobacteria Protein crystallization Protein folding Protein purification Protein solubility