Construction of an expression vector with elastin-like polypeptide tag to purify xylanase

Abstract:

Abstract:［Objective］This paper reports the purification of xylanase using the shortest，sensitive ELP [KV8 F-20]［Methods］We designed a thermophilic xylanase gene，and recombined it with the ELP via a random coil sequence to generate the vector pET-22b-SoxB-M2-S-ELP. The expressed xylanase was purified by inverse transition cycling through high-speed centrifugation，and then we characterized the purified xylanase．［Results］The phase transition temperature of the ELPs dropped to 22℃ with 0.5 mol/L sodium carbonate (pH=7)．Under this condition，SoxB-M2-S-ELP was purified by 3.16 folds after centrifugation． The recovery rate was 21.2%，and purity of the xylanase was 64.3%.［Conclusion］Elastinlike polypeptide as a purification tag to purify recombinant proteins is simple，fast，gentle and cheaper．The expression vector we constructed here might be a very useful and reliable tool to purify many other target proteins.