β-1,3-xylan is an important structural polysaccharide distributed in seaweeds including Porphyra.Polysaccharide-binding proteins are critical tools in polysaccharide research, serving as probes for the in situ analysis of polysaccharides.Carbohydrate-binding module (CBM) is a class of widely used polysaccharide-binding proteins.In the present study, a potential β-1,3-xylan binding sequence from the CBM6 family was discovered from the public gene database.The pure protein, named VaCBM6, was obtained by gene cloning, heterologous expression, and affinity purification.VaCBM6 exhibited a specific binding capacity to β-1,3-xylan, with an association constant of 3.0×105 (mol/L)-1.Furthermore, the first specific fluorescent probe for β-1,3-xylan was constructed by fusing VaCBM6 with the green fluorescent protein EmGFP, based on which the fluorescence microscopic observation of β-1,3-xylan in Porphyra was realized.The potential of VaCBM6 in the in situ analysis of β-1,3-xylan was confirmed.The discovery and characterization of VaCBM6 provided a promising tool for the future investigation and application of β-1,3-xylan.
JIANG Xiaoxiao
,
MEI Xuanwei
,
LIU Guanchen
,
ZHANG Yuying
,
SUN Haitao
,
CHEN Huanhuan
,
CHANG Yaoguang
,
XUE Changhu
. Cloning, expression, characterization, and application of a β-1,3-xylan-specific binding protein[J]. Food and Fermentation Industries, 2024
, 50(17)
: 11
-16
.
DOI: 10.13995/j.cnki.11-1802/ts.037509
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