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Volume 166, Issue 6

MSMEG_2432 of mc155 is a dual function enzyme that exhibits DD-carboxypeptidase and β-lactamase activities Free

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.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): cell shape , DD-carboxypeptidase , MSMEG_2432 , Mycobacterium and β-lactamase
Funding
This study was supported by the:
  • Department of Biotechnology, Government of India (Award BT/PR24255/NER/95/716/2017)
    • Principle Award Recipient: Anindya S Ghosh
© 2020 The Authors
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2020-04-17
2024-04-18
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MSMEG_2432 of Mycobacterium smegmatis mc2155 is a dual function enzyme that exhibits DD-carboxypeptidase and β-lactamase activities
Microbiology 166, 546 (2020); https://doi.org/10.1099/mic.0.000902
/content/journal/micro/10.1099/mic.0.000902
/content/journal/micro/10.1099/mic.0.000902
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