The Effects of N-glycosylation on Enzymological Properties of Phytase Phy(QF)

doi:10.7668/hbnxb.2015.01.030

Abstract

Abstract: In order to study the effects of N-glycosylation on enzymological properties of phytase phy(Q_F), the recombinant phytase gene phy(Q_F) was expressed in pichia pastoris GS115.Phytase phy(Q_F) was deglycosylated by endoglycosylase Endo H_f.Enzymology properties of glycosylated phy(Q_F) and deglycosylated phy(Q_F) were analyzed.The results showed that the glycosylated phy(Q_F) had a specific activity of 3 201 U/mg with an MW of approximately 53 kDa and the K_m and V_max of glycosylated phy(Q_F) were 392 μmol/L and 3 267 U/mg, respectively.The optimal pH value of glycosylated phy(Q_F) was 4.5 and was consistented with that of deglycosylated phy(Q_F).However, the optimal temperature of glycosylated phy(Q_F) was 55 ℃, 5 ℃ higher than that of deglycosylated phy(Q_F) and 2 ℃ increased in the melting temperature (T_m).The glycosylated phy(Q_F) exhibited 13% residual activity after treatment at 70 ℃ for 10 min and deglycosylated phy(Q_F)was completely deactivated under the condition.As one of posttranslational modifications, N-glycosylation may play an important role in improving the stability of phytase phy(Q_F).

Key words: Phytase, N-glycosylation, Pichia pastoris, Thermal stability, Circular dichroism

CLC Number:

WANG Xi, SONG Ma-li, DU Jun, FU Yue-jun, HU Feng-yun, LIANG Ai-hua. The Effects of N-glycosylation on Enzymological Properties of Phytase Phy(Q_F)[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(1): 177-181. doi: 10.7668/hbnxb.2015.01.030.

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