Cloning and expression of two novel β-glucosidase genes from Rhizopus stolonifer var.reflexus TP-02 cDNA library in Pichia pastrois
A n expressioncD N A library was constructed from Rhizopus stolonifer var. reflexes TP-02 with high capacity for hydrolysis of celluase , and then two novel β-glucosidase genes bgl1 and bgl2 were isolated and efficiently expressed in Pichia pastrois. The recom bined Pichia pastrois were culrured in the M M medium,using l℃ methanol to induce the expression of recom binant gene . The results showed that the maxinum activity of recom bined β-glucosidases were obtained at 84 h , 12 h earlier than that of the onginal strain Rhizoupus stolonifer var. reflexus T P-02 , and were 8 .2 IU /m L and 9 .9 IU /m L , 1.0-fold and 1.41-fold higher than that from the original strain . The exprPssed protein were purified from the ferm ented supernatant using Sephadex G 100 column and determ ined by SDS-PAGE . The results of SD S-PA G E also showed that the molecular weights of these two novel enzym es were 46 kD a and 47 kD a, respectively .
国家科技期刊平台
NSTL
万方数据
维普
