FIGURE 4.

In vitro activity of mutant CsBGlu12 proteins. Two mutant proteins designated as M1 (Glu²⁰⁰–Ala²⁰⁰) and M2 (Glu⁴¹⁴–Ala⁴¹⁴) were generated, and their activity was determined against cellobiose. A, TLC of wild type (WT) and mutant CsBGlu12. Lane L1, WT protein plus cellobiose; lane L2, cellobiose only; lane L3, M1 plus cellobiose; and lane L4, M2 plus cellobiose. G and C2 depict glucose and cellobiose, respectively. B, homology models of WT active site. Pink color, residues depict two catalytic Glu residues, M1 active site (C) and M2 active site (D). Red color residues depict mutated residues.