Reconstruction of diaminopimelic acid biosynthesis allows characterisation of Mycobacterium tuberculosis N-succinyl-L,L-diaminopimelic acid desuccinylase

Research output: Contribution to journalArticle

Authors

  • Adrian J Lloyd
  • David I Roper
  • Christopher G Dowson
  • G Kozlov
  • Kalle Gehring
  • Smita Chauhan
  • Hasan Iman
  • Claudia Blinder

Colleges, School and Institutes

External organisations

  • Department of Life Sciences, University of Warwick, Coventry, CV4 7AL, United Kingdom.

Abstract

With the increased incidence of tuberculosis (TB) caused by Mycobacterium tuberculosis there is an urgent need for new and better anti-tubercular drugs. N-succinyl-L,L-diaminopimelic acid desuccinylase (DapE) is a key enzyme in the succinylase pathway for the biosynthesis of meso-diaminopimelic acid (meso-DAP) and L-lysine. DapE is a zinc containing metallohydrolase which hydrolyses N-succinyl L,L diaminopimelic acid (L,L-NSDAP) to L,L-diaminopimelic acid (L,L-DAP) and succinate. M. tuberculosis DapE (MtDapE) was cloned, over-expressed and purified as an N-terminal hexahistidine ((His)6) tagged fusion containing one zinc ion per DapE monomer. We redesigned the DAP synthetic pathway to generate L,L-NSDAP and other L,L-NSDAP derivatives and have characterised MtDapE with these substrates. In contrast to its other Gram negative homologues, the MtDapE was insensitive to inhibition by L-captopril which we show is consistent with novel mycobacterial alterations in the binding site of this drug.

Details

Original languageEnglish
Article number23191
JournalScientific Reports
Volume6
Publication statusPublished - 15 Mar 2016