Abstract:

Gene PH1704 from an archeaon Pyrococcus horikoshii OT3 was cloned and expressed at high levels in E.coli BLPCodonPlus. The recomb
inant enzyme was purified to homogeneity via ultransonic, heating and HiTrap Q Sepharose chromatography. Through SDSPAGE, NativePAGE, activitystaining and westernblotting, the protease was identified as homooligomers with dodecamer being the major polymer, and showed resistance to SDS. The enzyme was characterized for azogelation, its optimum temperature and pH were 85 ℃ and 8.0, respectively. The enzyme had a favourable heat resistance.

Key words: thermophilic protease, expression, purification, character