Fig. 4: Biochemical characterization of selected identified enzymes.
a Phchr2|126075 was heterologously produced in K. lactis and activity was confirmed against pNP-acetate by following the release of pNP with a Vmax of 41.7 U mg−1 protein and a KM of 0.67 mM. b Substrate specificity of Phchr2|2915237 and WP_074995790 from S. misionensis. The activity with different p-nitrophenol-based substrates was determined by measuring the release of pNP. The activity on complex polysaccharides was determined via the DNSA assay that quantifies the formation of reducing ends upon polysaccharide cleavage. Phchr2|2915237 displayed the highest activities with pNP-Glc, pNP-Xyl, lichenan, and beech wood xylan, while WP_074995790, a close homolog of Phchr2|3002168 showed activity only with pNP-Gal as a substrate with a specific activity of 0.85 U mg−1 protein. c Kinetic characterization of Phchr2|2915237 using pNP-Glc, pNP-Xyl, lichenan, and beech wood xylan as substrates. All activity measurements were performed in triplicate (n = 3), mean values are shown and the error bars indicate the standard deviation (SD).
