Table 1.
QacA mutation | QacA expression (% wt QacA)c | minimum inhibitory concentration (μg ml-1)a.b | |||||
monovalent substrates | bivalent substrates | ||||||
dyes | Qacsd | Bg | Di | ||||
Et | PyY | Bc | Dc | Ch | Dd | ||
wt QacA | 100 | 1200 | 500 | 50 | 350 | 12 | 300 |
vector | N/A | 400 | 50 | 30 | 100 | 2 | 100 |
Y63V | 86 | 200 | 50 | 30 | 150 | 4 | 100 |
Y63F | 50 | 1400 | 400 | 50 | 250 | 12 | 300 |
Y315A | 64 | 800 | 500 | 50 | 300 | 12 | 400 |
Y315F | 75 | 1000 | 500 | 50 | 300 | 12 | 300 |
Y358S | 80 | 600 | 400 | 50 | 100 | 8 | 200 |
Y358F | 34 | 1400 | 400 | 50 | 300 | 16 | 300 |
Y366A | 106 | 1200 | 300 | 50 | 100 | 12 | 300 |
Y366V | 50 | 1200 | 400 | 50 | 300 | 12 | 300 |
Y366F | 99 | 800 | 500 | 50 | 300 | 12 | 400 |
Y410A | 132 | 400 | 50 | 45 | 100 | 3 | 100 |
Y410V | 67 | 200 | 50 | 30 | 100 | 7 | 300 |
Y410F | 58 | 200 | 100 | 50 | 250 | 12 | 300 |
Y429S | 57 | 200 | 200 | 30 | 250 | 4 | 100 |
Y429F | 32 | 1400 | 500 | 60 | 300 | 14 | 400 |
Y501V | 36 | 800 | 500 | 50 | 300 | 12 | 300 |
Y501F | 47 | 1200 | 500 | 50 | 350 | 12 | 400 |
a MIC values were determined in E. coli BHB2600 cells expressing wild-type QacA or a mutant derivative and represent the lowest concentration of substrate required to fully inhibit bacterial growth.
b MIC experiments were conducted in at least triplicate and the median value is presented.
c QacA protein expression levels are the average of two repeat experiments.
d Abbreviations: Qacs, quaternary ammonium compounds; Bg, biguanidines; Di, diamidines; Et, ethidium; PyY, pyronin Y; Bc, benzalkonium; Dc, dequalinium; Ch, chlorhexidine; Dd, diamidinodiphenylamine.