TABLE 1.
β-Glucosidase activities of selected P. pastoris GS115 transformants in shake flasks.
| Selected transformants | β-Glucosidase activities (U/L × 104) | Average enzymatic activity (U/L × 104) | G418 concentrations (mg/mL) |
| s1 | 10.8 ± 0.3 | 10.7 ± 0.9 (n = 3) | 2.5 |
| s2 | 9.8 ± 0.2 | ||
| s3 | 11.6 ± 0.3 | ||
| s4 | 9.7 ± 0.3 | 9.5 ± 0.9 (n = 5) | 2.0 |
| s5 | 10.4 ± 0.2 | ||
| s6 | 10.1 ± 0.2 | ||
| s7 | 8.2 ± 0.4 | ||
| s8 | 9.1 ± 0.2 | ||
| s9 | 7.1 ± 0.2 | 7.5 ± 0.6 (n = 5) | 1.5 |
| s10 | 8.3 ± 0.3 | ||
| s11 | 6.9 ± 0.3 | ||
| s12 | 7.5 ± 0.1 | ||
| s13 | 7.8 ± 0.2 | ||
| s14 | 6.1 ± 0.2 | 5.2 ± 0.9 (n = 5) | 1.0 |
| s15 | 3.8 ± 0.1 | ||
| s16 | 5.7 ± 0.2 | ||
| s17 | 5.0 ± 0.1 | ||
| s18 | 5.6 ± 0.2 | ||
| s19 | 2.4 ± 0.1 | 2.9 ± 0.6 (n = 5) | 0.5 |
| s20 | 3.1 ± 0.2 | ||
| s21 | 2.9 ± 0.4 | ||
| s22 | 2.7 ± 0.3 | ||
| s23 | 3.6 ± 0.2 | ||
| s24 | 0.77 ± 0.06 | 0.8 ± 0.2 (n = 5) | 0.1 |
| s25 | 0.69 ± 0.05 | ||
| s26 | 1.26 ± 0.3 | ||
| s27 | 0.74 ± 0.1 | ||
| s28 | 0.69 ± 0.06 | ||
| GS115 | ∼0 | N.A | 0 |
| GS115-3.5K | ∼0 | N.A | 0.1 |
Twenty-eight transformants were selected from YPD-agar plates containing 0.1, 0.5, 1.0, 1.5, 2.0, and 2.5 mg/mL G418. GS115 and GS115-3.5K indicate controls of the parental strain GS115 and the strain harboring empty pPIC3.5K vector, respectively. The β-glucosidase activity of the parental strain was deducted before calculating the enzymatic activity of recombinant FBG1. Results reflect the maximal values during entire 120 h induction. Experiments were done in triplicates. Three parallel measurements for each transformant were assayed. Values are indicated by means ± S.D. N.A indicates not available.