Purification and Kinetic Characterization of the Essential Condensation Enzymes Involved in Prodiginine and Tambjamine Biosynthesis. (3rd December 2019)

• Record Type: Journal Article
• Authors: Picott, Katherine J.; Deichert, Julie A.; deKemp, Ella M.; Snieckus, Victor; Ross, Avena C.
• Is Part Of: Chembiochem. Volume 21:Number 7(2020)
• Page Start: 1036
• Page End: 1042
• Publication Date: 2019-12-03
• Subjects: biosynthesis -- enzyme kinetics -- natural products -- prodiginine -- tambjamine; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.201900503
• Languages: English
• ISSNs: 1439-4227

Abstract

Abstract: Prodiginines and tambjamines are related families of bioactive alkaloid natural products with pharmaceutical potential. Both compound families result from a convergent biosynthetic pathway ending in the condensation of a conserved bipyrrole core with a variable partner. This reaction is performed by unique condensation enzymes, and has the potential to be manipulated to produce new pyrrolic compounds. We have purified and reconstituted the in vitro activity of the condensation enzymes PigC and TamQ from Pseudoalteromonas sp., which are involved, respectively, in the prodiginine and tambjamine biosynthetic pathways. Kinetic analysis confirmed a Uni Uni Bi Uni ping‐pong reaction sequence with competitive and uncompetitive substrate inhibition for PigC and TamQ respectively. The kinetic parameters of each enzyme provide insight into their differing substrate scope, and suggest that TamQ may have evolved a wide substrate tolerance that can be used for the production of novel prodiginines and tambjamines. Abstract : Bioengineering boost : Prodiginine and tambjamine condensation enzymes, PigC and TamQ, catalyze analogous reactions that form differing bond connectivity. Purification and kinetic analysis reveal that both enzymes follow a Ter Bi ping‐pong reaction profile with substrate inhibition. Their distinct substrate preferences suggest diverging evolution of these natural product biosynthetic gene clusters.