Abstract: Xylanase is one of the most important hydrolyzed enzymes with multiple functions in industry processing. Since there has been limited research related to β-1,3-xylanse, more attention needs to be paid to discovering an efficient way to produce this enzyme. In the present study, β-1,3-xylanase gene (xylII) from Pseudomonas vesicularis MA103 was cloned into pET-39b(+) expression vector and transformed into Escherichiacoli ClearColi BL21(DE3). The catalytic domain of the β-1,3-xylanase (XYLII) belongs to family 26 of glycoside hydrolases, and is followed by two family 31 carbohydrate-binding modules at the C terminus. β-1,3-xylanase showed an optimal activity at 35 °C and pH 7.5. According to its substrate specificity, the purified XYLII was considered to be an endo-type β-1,3-xylanase (EC 18.104.22.168). Experimental results of this work suggested an efficient way to obtain β-1,3-xylanase with great potential in industry applications. These findings can be helpful to explore and study the use of β-1,3-xylanase in the future and further associated investigation.