TABLE 1.
NO uptake by recombinant E. coli and M. smegmatis expressing glycosylation-deficient mutant of Mtb HbN
Cultures of E. coli and M. smegmatis mc2 155 were grown in vigorously aerated LB and Middlebrook 7H9 (Difco) medium, respectively, and harvested at the late exponential phase of growth when the culture A600 reached 0.8. Cells were washed with NO consumption buffer as described previously (10) and resuspended in the same buffer at a density of 5 × 107 cells/ml. NO consumption activity of cells was checked by NO microelectrode (WPI) following the procedure described previously (10). Data represent the mean of three independent results.
| Strains | Heme content | NO consumption activity |
|---|---|---|
| pmol/1010 cells | nmol−1 1010 cells | |
| E. coli BL21DE3 | 3.4 ± 0.9 | 0.05 ± 0.0 |
| E. coli BL21DE3 (HbN Wt) | 34.7 ± 4.8 | 17.84 ± 4.1 |
| E. coli BL21DE3 (HbNT132A,T133A) | 39.1 ± 5.6 | 19.4 ± 3.5 |
| M. smegmatis | 2.6 ± 1.1 | 0.11 ± 0.06 |
| M. smegmatis (HbN WT) | 24.6 ± 3.8 | 36.8 ± 4.1 |
| M. smegmatis (HbNT132A,T133A) | 29.4 ± 3.3 | 20.9 ± 2.9 |