Table 1 Expression of the BCO forms using different growth and induction conditions.
From: Production of recombinant cholesterol oxidase containing covalently bound FAD in Escherichia coli
Phase of induction | Plasmid (BCO form) | Time after induction (h) | Temperature after induction | |||
|---|---|---|---|---|---|---|
25°C | 37°C | |||||
U/mgprotein | U/Lculture | U/mgprotein | U/Lculture | |||
Exponential phase (OD600 nm~1) | pET-fBCO (fBCO) | 0 | b.d. | b.d. | b.d. | b.d. |
3 | ≤ 0.01 | < 1.0 | ≤ 0.01 | < 1.0 | ||
7 | 0.07 ± 0.005 | 2.0 ± 0.3 | 0.10 ± 0.05 | < 1.0 | ||
18 | 0.16 ± 0.05 | 260 ± 80 | 0.12 ± 0.02 | 1.8 ± 0.2 | ||
pET-mBCO (mBCO) | 0 | b.d. | b.d. | b.d. | b.d. | |
3 | 0.31 ± 0.02 | 130 ± 10 | 0.09 ± 0.02 | 50 ± 11 | ||
7 | 0.38 ± 0.14 | 190 ± 50 | 1.0 ± 0.3 | 390 ± 45 | ||
18 | 0.54 ± 0.15 | 500 ± 120 | 0.17 ± 0.06 | 240 ± 20 | ||
Stationary phase (OD600 nm~2) | pET-fBCO (fBCO) | 0 | b.d. | b.d. | b.d. | b.d. |
3 | ≤ 0.01 | < 1.0 | ≤ 0.01 | < 1.0 | ||
7 | ≤ 0.01 | 1.1 ± 0.1 | ≤ 0.01 | < 1.0 | ||
18 | 0.03 ± 0.015 | 2.5 ± 0.4 | 0.02 ± 0.01 | 1.8 ± 0.2 | ||
pET-mBCO (mBCO) | 0 | b.d. | b.d. | b.d. | b.d. | |
3 | 0.52 ± 0.12 | 400 ± 10 | 0.22 ± 0.06 | 150 ± 35 | ||
7 | 0.28 ± 0.02 | 180 ± 20 | 0.55 ± 0.18 | 180 ± 10 | ||
18 | 0.13 ± 0.02 | 110 ± 30 | 0.12 ± 0.05 | 85 ± 9 | ||