Subsequent to this, Alvarez and Tavel found organisms similar to that described by Lustgarten also in normal genital secretions (smegma).

Essentially, the bacteria form a single-layered sheet and are able to move slowly together without the use of any extracellular structures, like flagella or pili.

Although it hasn’t been determined exactly how this mechanism works, the surface properties of the unique cell wall (Figure 1) of M. smegmatis have been found to play a role.

[9][10][11][12] The discovery of plasmids, phages, and mobile genetic elements has enabled the construction of dedicated gene-inactivation and gene reporter systems.

M. smegmatis has three options for repairing double-strand breaks; homologous recombination (HR), non-homologous end joining (NHEJ), and single-strand annealing (SSA).

[16] The HR pathway of M. smegmatis is the major determinant of resistance to ionizing radiation and oxidative DNA damage.

[17] SSA is employed as a repair pathway when a double-strand break arises between direct repeat sequences in DNA.

SSA involves single-strand resection, annealing of the repeats, flap removal, gap filling and ligation.

M. smegmatis is a simple model that is easy to work with, i.e., with a fast doubling time and only requires a biosafety level 1 laboratory.

[citation needed] Mycobacterium smegmatis shares the same peculiar cell wall structure of M. tuberculosis and other mycobacterial species.

M. smegmatis is commonly used to study ESX secretion because of its genetic similarities  and analogous function to M. tuberculosis, as well as ease of growing in the lab.

[20] Mycobacterium smegmatis is readily cultivatable in most synthetic or complex laboratory media, where it can form visible colonies in 3–5 days.

In 2023, researchers reported extracting from M. smegmatis a hydrogenase called Huc, which is highly efficient at oxidizing hydrogen gas—and thus creating an electric current—while also being insensitive to the presence of oxygen, which typically obstructs catalysis.