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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
  1. In all cases, E. coli cells carrying the recombinant pET24b(+)-BCO plasmids were cultivated in LB medium and protein expression was induced by 1 mM IPTG addition. b.d., below detection limit. Data are expressed as mean ± S.D., n = 3.