Effects of different media supplements on the production of an active recombinant plant peroxidase in a Pichia pastoris Δoch1 strain

Figures & data

Figure 1. The recombinant Δoch1 strain was cultivated under inducing conditions in the presence of different heme precursors in shake flasks at 25°C for 96 hours. (A) OD₆₀₀ values over induction time; (B) volumetric enzyme activity in U·mL⁻¹ over induction time. Dark gray solid line with triangles, no heme precursor; light gray solid line with squares, ALA; dark dashed line with diamonds, FeSO₄; back solid line with dots, hemin.

Table 1 Values for OD₆₀₀, HRP activity, total protein content and specific activity for the recombinant Δoch1 strain induced in the presence of different heme precursors for 96 hours

Heme precursor	OD600	HRP activity [U· mL^−1]	Protein content [mg· mL^−1]	Specific activity [U· mg^−1]
No precursor	8.81	4.79	0.056	84.8
ALA [1 mM]	8.57	4.51	0.053	85.5
FeSO4 [1 mM]	8.99	7.70	0.065	117.9
hemin [30 μM]	8.59	28.1	0.101	278.1

Figure 2. A recombinant benchmark strain and the recombinant Δoch1 strain were cultivated under inducing conditions in the presence or absence of a protease inhibitor cocktail in shake flasks at 25°C for 96 hours. (A) OD₆₀₀ values over induction time; (B) total extracellular protein concentration in mg·mL⁻¹ over induction time. Dark gray solid line with squares, benchmark strain without protease inhibitor; dark gray dashed line with dots, benchmark strain with protease inhibitor; black solid line with squares, Δoch1 strain without protease inhibitor; black dashed line with squares, Δoch1 strain with protease inhibitor.