An effective combination of codon optimization, gene dosage, and process optimization for high-level production of fibrinolytic enzyme in Komagataella phaffii (Pichia pastoris)

Table 4 Effects of metal ions on fibrinolytic enzyme activity

Metal ions	Concentration relative activity (%)^a
Metal ions	5 mM	20 mM	50 mM
Ca²⁺	117.5 ± 1.8	121.2 ± 2	97.1 ± 1.2
Mg²⁺	100.5 ± 1	108.9 ± 1.4	98.0 ± 2.1
Zn²⁺	8.1 ± 1.4	4.5 ± 1.2	1.8 ± 1.0
Cu²⁺	5.4 ± 1.9	0.0 ± 1.0	0
Mn²⁺	88.3 ± 1.6	7.3 ± 1.4	0
Pb²⁺	0	0	0
Fe³⁺	0	0	0
CK^b	100	100	100

^aThe relative activity with different metal ions added was determined and compared with the activity measured in phosphate buffer (pH 7.4, 0.02 M) without the addition of any ions
^bThe relative activity measured in phosphate buffer (pH 7.4, 0.02 M) without the addition of any ions
All values are mean ± SD from three independent experiments

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