1887

Abstract

Mycobacterial peptidoglycan (PG) is an unsolved puzzle due to its complex structure and involvement of multiple enzymes in the process of its remodelling. -Carboxypeptidases are low molecular mass penicillin-binding proteins (LMM-PBPs) that catalyzes the cleavage of terminal -Ala of muramyl pentapeptide branches and thereby helps in the PG remodelling process. Here, we have assigned the function of a putative LMM-PBP, MSMEG_2432 of , by showing that it exhibits both -CPase and β-lactamase activities. Like conventional -CPase (PBP5 from ), upon ectopic complementation in a deformed seven PBP deletion mutant of , MSMEG_2432 has manifested its ability to restore ~75 % of the cell population to their normal rod shape. Further, -CPase assay has confirmed its ability to release terminal -Ala from the synthetic tripeptide and the peptidoglycan mimetic pentapeptide substrates ending with -Ala--Ala. Also, elevated resistance against penicillins and cephalosporins upon ectopic expression of MSMEG_2432 suggests the presence of β-lactamase activity, which is further confirmed through nitrocefin hydrolysis assay. Moreover, it is found apparent that D169A substitution in MSMEG_2432 influences both of its and -CPase and β-lactamase activities. Thus, we infer that MSMEG_2432 is a dual function enzyme that possesses both -CPase and β-lactamase activities.

Funding
This study was supported by the:
  • Anindya S Ghosh , Department of Biotechnology, Government of India , (Award BT/PR24255/NER/95/716/2017)
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/content/journal/micro/10.1099/mic.0.000902
2020-04-17
2020-06-04
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