Identification and characterization of a thermostable GH11 xylanase from Paenibacillus campinasensis NTU-11 and the distinct roles of its carbohydrate-binding domain and linker sequence

An extracellular thermostable xylanase (XynNTU) from Paenibacillus campinasensis NTU-11, consisted of a glycoside hydrolase (GH) family 11 catalytic domain, a Gly/Pro-rich linker sequence (LS) and a family 6 carbohydrate-binding module (CBM6), was identified and expressed in E. coli BL21. The purified XynNTU had a specific activity of 2750 U/mg and an optimal activity at 60 degrees C and pH 7.0, and retained a residual activity of 58.4% after incubation (60 degrees C, 48 h). Two truncated mutants, CBM6-truncated form XynNTU-CDLS, CBM6 and linker-truncated form XynNTU-CD, possessed similar values of optimum pH and temperature as the native XynNTU. XynNTU-CD displayed a lower thermostability than XynNTU, whereas for XynNTU-CDLS, more than 90% of residual activity was remained (60 degrees C, 48 h), indicating that this enzyme presented a higher thermo-stability than that of the majority of reported GH11 xylanases. Furthermore, XynNTU and two mutants main-tained more than 70% of residual activity at pH values of 5-9. Kinetic measurements suggested that CBM6 had a crucial function in the ability of the enzyme to bind and hydrolyze xylan substrates, while LS had a relatively mild influence. Collectively, a noticeable thermostability and a high specific activity of XynNTU and its truncated form XynNTU-CDLS highlights their potentials for diverse industrial applications.

Automated summary

The extracellular thermostable xylanase (XynNTU) from Paenibacillus campinasensis NTU-11 displayed high thermal stability and specific activity, with truncated mutants showing similar optimal conditions. The carbohydrate-binding module (CBM6) was crucial for enzyme-substrate binding and hydrolysis, highlighting the potential industrial applications of XynNTU and its mutants.