Keywords

Wheat Xylanase Xylanase Inhibitory Protein Inhibitory Properties

Abstract

[Objective] Purify wheat xylanase inhibitory protein from mature wheat grains and study its inhibitory effect on xylanase, so as to explore the effect of xylanase inhibitory protein in wheat on external xylanase. [Method] Xylanase inhibitory enzyme was isolated and purified from Xiaoyan 22 wheat using ammonium sulfate salting out, Macro-prep CM cation exchange chromatography, Macro-prep DEAE anion exchange chromatography and Macro-prep 25S cation exchange chromatography. Protein, the purity and molecular mass of xylanase inhibitory protein were detected by SDS-PAGE electrophoresis, and the effects of different temperatures and reaction times on the inhibitory activity of xylanase inhibitory protein were further studied. [Results] The molecular weight of wheat xylanase inhibitory protein is about 29 ku, and the N-terminal amino acid sequence is SVSSVVS. After NCBI BLAST comparison, the N-terminal sequence of this inhibitory protein has no homology with other xylanase inhibitory proteins, but It is highly homologous to the N-terminal sequence of barley chitinase, but has no chitinase activity; this protein has strong inhibitory activity against the GH11 family xylanase from Aspergillus niger, but has no activity against the GH10 family xylanase. Sensitive; the optimal temperature for the protein to bind to xylanase is 30¡ãC, and the optimal time is 30 min; the half-inactivation temperature of the inhibitory protein is 74¡ãC, and it has good thermal stability. [Conclusion] The obtained wheat xylanase inhibitory protein may be a new type of XIP-type xylanase inhibitory protein

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