A novel, alkali-tolerant thermostable xylanase from Saccharomonospora viridis: direct gene cloning, expression and enzyme characterization.

A xylanase gene, designated Svixyn10A, was cloned from actinomycetes Saccharomonospora viridis and the gene product was characterized. Gene Svixyn10A contains 1,374 bp and encodes a polypeptide of 457 amino acids composed of a glycoside hydrolase family 10 catalytic domain with a putative signal peptide, a short Gly-rich linker and a family 2 carbohydrate-binding module (CBM). The deduced amino acid sequence of SviXyn10A shared the highest identity (57 %) with a hypothetical xylanase from Streptomyces lividans TK24 (ZP_05528201). A recombinant His-tagged xylanase, SviXyn10A was expressed in Escherichia coli BL21 and purified. The optimum pH and temperature for SviXyn10A is 8.0 and 60 °C. Compared with thermophilic and mesophilic counterparts, SviXyn10A was more active at high temperatures, retaining >63 % of its maximum activity at 65-70 °C and ~40 % even at 80 °C. It had broad pH adaptability (>35 % activity at pH 5.0-11.0) and alkali-tolerance (>70 % activity after incubation at pH 8.0-11.0 for 1 h at 37 °C), and was highly thermostable (>75 % activity after incubation at 70 °C for 3 h at pH 8.0). It may be the first alkali-tolerant thermostable xylanase reported from Saccharomonospora. These favorable properties make SviXyn10A a good candidate for application in pulp and paper industries.