Characterization of endogenous carbohydrates extracted from recombinant enzyme preparations.

Recombinant proteins were denatured and treated with amyloglucosidase prior to glucose estimation. Heterologous expression were either performed in E. coli BL21(DE3) strains or in glycogen deficient ΔglgCAP cells with the same genetic background. For characterization of endogenous glucans enzyme preparations expressed in BL21(DE3) were denatured and treated with isoamylase. Debranched products were analysed by HPAEC-PAD. For evaluation of data different chain length peaks were grouped. Only DP populations (DP2–7, DP8–13, and DP14–20) were tested for significance among the enzyme preparations.
Differences were significant with * P = 0.05, ** P = 0.02 or *** P = 0.01 (rSSI with rBE2 and rBE3). Differences (DP-2-7 and DP8–13) between rBE2 and rBE3 were significant with P = 0.05. Average values and standard deviation of four independent experiments for each preparation are given. a) Ratio between nmol glucose and nmol protein. n.d. – not detectable.