Table 1 Comparison of enzyme production and purification for the four expression hosts

Enzyme production

 Cultivation time (h)

28

111

288

288

 Wet cell mass (g L⁻¹)

4785 ± 160

275

n.d.

n.d.

 Soluble protein conc. (mg L⁻¹)

1517 ± 44^a

633

108 ± 4

94 ± 3

 DCIP activity (U L⁻¹)

648 ± 19

376

49 ± 2

715 ± 14

 Cyt c activity (U L⁻¹)

–

320

21 ± 1

362 ± 11

Enzyme purification

 Purification steps

IMAC

Phenyl Sepharose and QSource

Phenyl Sepharose and QSource

Phenyl Sepharose and QSource

 Purification yield (%)

60

71

54

58

 Purification (fold)

65.5

7.7

51.8

3.4

 Specific activity for DCIP (U mg⁻¹)

27.5 ± 0.3

9.4 ± 0.2

14.1 ± 0.1

14.3 ± 0.1

 Specific activity for cyt c (U mg⁻¹)

–

3.5 ± 0.1

11.1 ± 0.2

12.5 ± 0.2

 FAD loading (%)

52 ± 2

44 ± 1

56 ± 1

68 ± 3

 Glycosylation (%)

0

18.9 ± 12.1

9.4 ± 2.7

1.4 ± 1.3