Table 1 Experimental design for optimization of CLEA-protease preparation parameters, using faced cantered central composite design FCCCD and response surface methodology RSM, all experiments were done in triplicate
Run | Glutaraldehyde (mM) | Acetone % (v/v) | BSA (mM) | Protease specific activity (U/mg protein) | CLEA-protease recovered activity (%)a |
|---|---|---|---|---|---|
1 | 65.00 (0) | 60.00 (+1) | 0.113 (0) | 3.03 | 22.04 |
2 | 80.00 (+1) | 60.00 (+1) | 0.188 (+1) | 1.61 | 11.71 |
3 | 50.00 (−1) | 60.00 (+1) | 0.188 (+1) | 3.01 | 21.89 |
4 | 65.00 (0) | 30.00 (−1) | 0.113 (0) | 2.85 | 20.73 |
5 | 65.00 (0) | 45.00 (0) | 0.113 (0) | 4.57 | 33.24 |
6 | 50.00 (−1) | 45.00 (0) | 0.113 (0) | 3.43 | 24.95 |
7 | 80.00 (+1) | 30.00 (−1) | 0.188 (+1) | 0.96 | 6.98 |
8 | 65.00 (0) | 45.00 (0) | 0.113 (0) | 3.63 | 26.40 |
9 | 80.00 (+1) | 30.00 (−1) | 0.038 (−1) | 1.13 | 8.22 |
10 | 65.00 (0) | 45.00 (0) | 0.113 (0) | 4.14 | 30.11 |
11 | 80.00 (+1) | 45.00 (0) | 0.113 (0) | 2.91 | 21.16 |
12 | 50.00 (−1) | 60.00 (+1) | 0.038 (−1) | 2.16 | 15.71 |
13 | 50.00 (−1) | 30.00 (−1) | 0.188 (+1) | 1.93 | 14.04 |
14 | 50.00 (−1) | 30.00 (−1) | 0.038 (−1) | 1.67 | 12.15 |
15 | 80.00 (+1) | 60.00 (+1) | 0.038 (−1) | 2.73 | 19.85 |
16 | 65.00 (0) | 45.00 (0) | 0.188 (+1) | 2.79 | 20.29 |
17 | 65.00 (0) | 45.00 (0) | 0.038 (−1) | 2.70 | 19.64 |