Molecular Cloning and Optimization for High Level Expression of Cold-Adapted Serine Protease from Antarctic Yeast Glaciozyma antarctica PI12
Figure 11
(a) Coomassie-stained SDS-PAGE of recombinant P. pastoris clone. 1 mL supernatants collected from the third-day culture were concentrated 10-fold using trichloroacetic acid, and 10 μL resultant products were applied to SDS-PAGE analysis. Lane 1: molecular weight marker (Fermentas); Lane 2: GS115 (Control); Lane 3: GS115/pPIC9 empty (Control); Lane 4: GS115/pPIC9/PI12prot (GpPro2). The arrow indicates the secreted recombinant PI12 protease. (b) Plate activity staining of different recombinant P. pastoris clones. G: GS115; GP: GS115/pPIC9; GRI: GS115/pPIC9/PI12prot1 (GpPro1); GRII: GS115/pPIC9/PI12prot2 (GpPro2); K: KM71; KP: KM71/pPIC9; KR: KM71/pPIC9/PI12prot (KpPro). (c) Gel activity staining of recombinant PI12 protease in P. pastoris GpPro2 clone. Lane 1: prestained molecular weight marker (Fermentas); Lane 2: GS115/pPIC9 empty (Control); Lane 3: GS115/pPIC9/PI12prot1 (GpPro2). The arrow indicates the secreted recombinant PI12 protease.
