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Fig. 2 | BMC Biotechnology

Fig. 2

From: Purification and biochemical characterization of a novel thermostable protease from the oyster mushroom Pleurotus sajor-caju strain CTM10057 with industrial interest

Fig. 2

Purification and identification of the protease SPPS from Pleurotus sajor-caju strain CTM 10057. a Chromatography profile of the partial protease purification on FPLC system using UNO Q-6. The column (12 mm × 53 mm) (Bio-Rad Laboratories, USA) was equilibrated with buffer C. The adsorbed material was eluted with a linear NaCl gradient (0 to 500 mM in buffer C at a flow rate of 30 mL/h, as described in Methods section. b The assessment of homogeneity and molecular weight analysis of purified SPPS protein on Native-PAGE, Lane 1, protein markers (molecular masses in kDa). Lane 2, purified SPPS enzyme from Pleurotus sajor-caju strain CTM 10057 (30 μg). c Chromatography profile of the purified protease SPPS on HPLC system using ZORBAX PSM 300 HPSEC. The column (6.2 mm × 250 mm) (Agilent Technologies, Lawrence, Kansas, MO, USA) was equilibrated with buffer C and native protein markers of 670, 158, 44, 17, and 13.5 kDa, show a single peak of 65 kDa, approximately. Proteins were separated by isocratic elution at a flow rate of 30 mL/h with buffer C and detected using a UV-VIS Spectrophotometric detector (Knauer, Berlin, Germany) at 280 nm. The pure SPPS enzyme, with retention time (Rt) of 11.6 min, contains protease activity. d SDS-PAGE 12% of the purified protease. Lane 1, Total cell extract. Lane 2, Empty. Lane 3, Purified SPPS (30 μg) obtained after HPLC-ZORBAX PSM 300 HPSEC chromatography at Rt = 11.6 min. Lane 4: Amersham LMW protein marker (GE Healthcare Europe GmbH, Freiburg, Germany). e Zymogram caseinolytic activity staining of the purified protease SPPS (30 μg)

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