PKS-NRPS Enzymology and Structural Biology: Considerations in Protein Production.
Methods Enzymol
; 604: 45-88, 2018.
Article
in En
| MEDLINE
| ID: mdl-29779664
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.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Peptide Synthases
/
Recombinant Proteins
/
Protein Engineering
/
Polyketide Synthases
Language:
En
Journal:
Methods Enzymol
Year:
2018
Document type:
Article
Affiliation country: