嗜热菌Pyrococcus horikoshiiOT3蛋白酶基因的原核表达、 纯化和性质表征

Abstract

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

CLC Number:

Q786

DAN Dong-Ling, BAI Ai-Xi, BAI He, HAN Wei-Wei, FENG Yan. Prokaryotic Expression, Purication and Properties of a Proteasefrom Thermophilic Archaeon Pyrococcus horikoshii OT3[J].J4, 2010, 48(06): 1065-1069.

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Prokaryotic Expression, Purication and Properties of a Proteasefrom Thermophilic Archaeon Pyrococcus horikoshii OT3

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