A thorough analysis and categorization of bacterial interrupted adenylation domains, including previously unidentified families

Abstract

In-depth study of intriguing bacterial interrupted adenylation domains from seven distinct families and six different types.

Lessons learned in engineering interrupted adenylation domains when attempting to create trifunctional enzymes from three independent monofunctional ones

“…This also sheds light as to why this is the most common location for interruptions of not only length of interruption, but type of M domain as well. 6,33 The two M domains that were added between the a8-a9 region were a total of 670 amino acids in length, and adenylation activity was still observed, indicating size may not be a serious constraint in engineering interrupted A domains. This study demonstrates the remarkable ability of the A domain to maintain the folding and positioning of the A subdomain, which is known to form the open and closed state of the A domain by moving in and out of the A domain binding pocket.…”

(S)‐3‐aminopiperidine‐2,6‐dione is a biosynthetic intermediate of microbial blue pigment indigoidine

“…The Ox domain of 3 synthetases like IdgS is embedded in the A domain, splitting the A domain into an N-terminal part (1-525 residues in IdgS) and a C-terminal part (864-929 residues in IdgS). It was shown previously that the function of the domains embedded in the A domain significantly relies on the integrity of NRPS modules, which emphasizes the difficulty in investigating IdgS-Ox as a single domain protein 35 . Therefore, we turned to study the dehydrogenation activity of IdgS-Ox in vitro using IdgS-TE* S1102A under different assay conditions (Figures 3B and S4).…”