Biofilm Associated Drug Target Identification of Mycobacterium Fortuitum

Abstract

Biofilm formation by Mycobacterium fortuitum increases the pervasiveness of nosocomial infections, leading to increased morbidity, mortality, and financial impact on healthcare systems. Molecular and phenotypic development of M. fortuitum biofilm has not been investigated in depth, requiring a study on identification of biofilm-associated drug targets and understanding of the related pathophysiology of M. fortuitum. The present study exploited Response Surface Methodology (RSM) to predict a combination of most optimum environmental conditions leading to maximum biofilm formation by M. fortuitum. The RSM-based statistical modeling showed that M. fortuitum formed biofilm with maximum thickness post 96 h of incubation, under static conditions, at 37°C, and a neutral pH. Literature review, homology search, along with the significance of mycobacterial fatty acid synthase type-II (FAS-II) components as attractive targets for designing anti-biofilm inhibitors, lead to short-listing of two novel FAS-II genes, MFfabG4 and MFhadC, in M. fortuitum, that were analyzed using antisense (knockdown) strategy. The gene knockdown studies confirmed and established MFfabG4 as a pellicle and biofilm specific marker for M. fortuitum. MFhadC, however, did not appear as a promising target for developing anti-biofilm drugs against M. fortuitum. A comparative global proteome analysis of M. fortuitum was performed under planktonic and biofilm-forming conditions. The differential global proteome profiling unraveled molecular bases underlying survival, and pathogenesis of M. fortuitum under biofilm growth conditions. The analysis also presented M. fortuitum planktonic and biofilm specific markers as potential drug targets. Molecular modeling was used to predict 3-D structures of M. fortuitum biofilm-specific markers that may be used for identification of their potential inhibitors. Collectively, the study helps identify and establish potential drug targets for anti-biofilm drug development against M. fortuitum.