纳豆激酶原基因在毕赤酵母中的分泌表达

Secreted expression of pro-nattokinase gene by Pichia psstoris

为构建纳豆激酶原基因的重组酵母菌分泌型表达载体pPRONK2,并在毕赤酵母菌中进行表达,将来源于豆豉的纳豆芽孢杆菌的纳豆激酶原基因克隆至毕赤酵母菌分泌型表达载体pHBM905A上,得到重组质粒pPRONK_2,再将其经SalⅠ酶切后分别转化3株毕赤酵母菌KM71、GS115、SMD1168,在MD平板上筛选得到重组酵母菌。经检测重组酵母菌发酵液中具纳豆激酶纤溶活性。表明在毕赤酵母菌中成功地实现了纳豆激酶的分泌表达。

In order to express pro-nattokinase gene in Pichia pastoris, a secreted-expression plasmid pPRONK2 harboring pro-nattokinase gene of Bacillus natto sawamura was constructed from pHBM905A. Then the plasmid pPRONK2 was digested by Sa/I and transformed into P. pastoris strains named KM71, GS115 and SMD1168, respectively. The recombinant P. pastoris sWains were isolated on MD plates. The fibrinolytic activity was detected in the fermentation broth of the recombinant yeasts. It was indicated that the secreted expression of nattokinase gene was realized successfully in P. Pastoris.