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Shivange et al.
Structure function relationships of Yersinia mollaretii phytase
120
Relative activity (%)
100
M6
Wt
M3
80
60
40
20
0
2
3
4
5
6
pH
7
8
9
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Fig. S10 Activity-pH profile of the Ymphytase wildtype and variant M3 and M6.
Native polyacrylamide gel electrophoresis (PAGE) of wildtype Ymphytase and variant M3
and M6
Native polyacrylamide gel (10%) was prepared by omitting SDS from the routinely used
polyacrylamide gel electrophoresis. NativeMark Unstained Protein Standard (Invitrogen) was used
as a molecular marker to estimate the quaternary form of Ymphytase variants. Purified protein of
Ymphytase variants [7 μL of protein and 3 μL of loading dye (tris 62.5 mM pH 6.8, glycerol 30%,
bromophenol blue 0.01%)] were loaded into each lane of the native gel. The electrophoresis was
performed using tris–glycine buffer (pH 8.3) and electrophoresis chamber was placed in an icewater bath during the run. Protein bands were stained with Coomassie brilliant blue solution.
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