The main outcomes of this study can be summarized as
Regarding the other isoenzymes especially HRP 04627 (C2), A2A
and 5508 could be purified very efficiently with PFs of 62, 684
and 144, respectively. HRP 25148.1 (C1C), 25148.2 (C1D),
04627 (C2), A2B and 1805 were purified 15- to 38-fold.
The correlation between the amount of potential N-glycosylation
sites and the success in flowthrough purification can be
used to design an efficient purification strategy for glycosylated
proteins expressed in P. pastoris in general.
Basic biochemical characterization using ABTS and TMB
revealed significant differences of the individual isoenzyme
preparations. The preparations of HRP A2A and HRP A2B turned
out to be highly active with H2O2 and ABTS and hence are especially
interesting for applications in diagnostic assays with high
sensitivity.