TABLE 3.
Strain | Analyzed parameter and condition
|
|||||||||
---|---|---|---|---|---|---|---|---|---|---|
ldhA mRNA
|
ackA mRNA
|
pta mRNA
|
ackA-lacZ (Miller units)
|
Pta activity (U/mg)d
|
||||||
Aerobic | Anaerobice | Aerobic | Anaerobic | Aerobic | Anaerobic | Aerobic | Anaerobic | Aerobic | Anaerobic | |
Wild type | 3.6 ± 0.14b | 2.8 ± 1.3b | 25.6 ± 1.2b | 142.8 ± 32b | 13.4 ± 3.0b | 90.8 ± 33b | 95 ± 7 | 355 ± 38 | 4.2 ± 0.5 | 24 ± 1.3 |
Δanr | NDc | 2.9 ± 1.5b | ND | 50.3 ± 4.1b | ND | 25.1 ± 6.1b | 90 ± 5 | 110 ± 13 | ||
dnr::tet | ND | 3.3 ± 1.5b | ND | 152 ± 0b | ND | 140 ± 18b | 104 ± 15 | 201 ± 14 | ||
ifhA::tet | 45 ± 6 | 90 ± 6 | ||||||||
narL::cat | 102 ± 7 | 290 ± 14 | ||||||||
ldhA::aacC1 | 4.3 ± 0.7 | 26 ± 1.8 | ||||||||
pta::aacC1 | 0.4 ± 0.2 | 0.5 ± 0.2 |
The results were obtained from microarray experiments and from a ackA-lacZ fusion. The Pta activity in P. aeruginosa wild type and the Δpta and ΔldhA mutants were determined as described in Materials and Methods.
Observed signal strength is given in normalized Affymetrix units as defined in Materials and Methods (5, 8). The standard deviation represents the detected differences between the independently performed GeneChip experiments.
ND, not determined. Because Anr and Dnr activity is known to be limited to anaerobic conditions, analysis of aerobically grown cells was not performed.
One unit of activity is the amount of enzyme that converts 1 μmol of acetyl-CoA to product per 60 min at 30°C.
Anaerobic conditions for the microarrays, the reporter gene fusions, and enzyme activity measurements differed depending on the nature of determined parameters. Due to the short half-life of the mRNAs, the RNA for the microarray analysis was prepared 2 h after the shift to anaerobic conditions. Reporter gene fusion expression was determined after 48 h of anaerobic incubation of the culture, while Pta activity was measured after 24 h of fermentative conditions. Employed media, test principles, and given units are defined in Materials and Methods.