Fig. 6

Collagen substrate degradation by recombinant collagenases. A SDS-PAGE analysis of gelatin digestion by recombinant collagenases during cell cultivation. Strains were cultivated in YPD medium with 10 mM CaCl₂, 0.6 mM ZnCl₂ and 1% gelatin at 30 °C for 96 h, then supernatant was used for analysis. Ref.: YPD medium with 10 mM CaCl₂, 0.6 mM ZnCl₂ and 1% gelatin. B Collagen hydrolysis by recombinant collagenases. Strains were cultivated in YPD medium with 10 mM CaCl₂ and 0.6 mM ZnCl₂ at 30 °C for 96 h for collagenase production. The supernatant was collected and replaced with 50 mM Tris–HCl (pH = 7.5) by using an Amicon^® Ultra filter (50 kDa). Subsequently, collagen was added to tubes and incubated at 37℃. G + H: supernatants from BG02 and BH01 were mixed with a ratio of 1:1. C Differences between recombinant ColG and ColH in collagen degradation were revealed at microscopic structure level by using a scanning electron microscope (SEM) with 20,000 × magnification. Day 1 degradation samples were used for SEM analysis