Characterization of yeast cell surface displayed Lentinula edodes xylanase and its effects on the hydrolysis of wheat

Abstract

The current study displayed a xylanase from Lentinula edodes on the surface of Pichia pastoris (sdLeXyn) and investigated its properties and effects on the wheat hydrolysis. Fluorescence microscope results showed that sdLeXyn was successfully anchored and displayed on the surface of P. pastoris X-33 cells. The highest activity of sdLeXyn was obtained at pH 3.0 and 50 °C. The sdLeXyn exhibited anti-high temperature property and showed broad temperature adaptability (>55% of the highest activity at 20-80 °C). The sdLeXyn was very stable at room temperature and could remain functionally stable at 50 °C for 3 h. The K_m value was greater in sdLeXyn than that in free recombinant L. edodes xylanase. The sdLeXyn exhibited well resistance to Mn²⁺, Zn²⁺, Ca²⁺, Na⁺, Cu²⁺, Mg²⁺, K⁺, Ni²⁺ (1 mM and 5 mM) except Cu²⁺, which reduced the sdLeXyn activity by 54.5% at 5 mM dosage. The activity of sdLeXyn was increased by 42.6% by 5 mM Mn²⁺, 5 mM DTT, Trition X-100, and Tween 20 did not affect the activity of sdLeXyn, but SDS and EDTA slightly reduced it by 12.8% and 14.6%, respectively. The sdLeXyn could resist the degradation of pepsin, efficiently hydrolyzed wheat and reduced the viscosity of wheat hydrolysate. Current data indicate that the sdLeXyn has a potential as a feed additive to improve the utilization of wheat in poultry production.

Keywords: Lentinula edodes xylanase; Surface display; Viscosity; Wheat.