Table 2.
Kinetic parameters of bone-degrading enzymes.
| Protein | Substrate | Km (mM)3 | Vmax (U mg−1)3 | kcat (s−1)3 | kcat/Km (mM−1 s−1)3 |
|---|---|---|---|---|---|
| T0182 | BODIPY-FL casein1 | n.d. | 125 ± 182 | n.d. | n.d. |
| FALGPA | 4.24 ± 0.1 | n.d. | 1.77 ± 0.30 | 0.42 ± 0.04 | |
| T0191 | pNP-Neu5Ac | 0.18 ± 0.03 | 6.35 ± 0.45 | 511.71 ± 32.71 | 2,955 ± 674 |
| 3-Sialyllactose | 0.031 ± 0.001 | n.d. | 921.5 ± 3.17 | 29,725 ± 15 | |
| 6-Sialyllactose | 0.053 ± 0.001 | n.d. | 550.5 ± 15.16 | 10,386 ± 20 | |
| T0193 | pNP-Neu5Ac | 0.55 ± 0.17 | 4.79 ± 0.79 | 508.05 ± 35.85 | 1,044 ± 388 |
| 3-Sialyllactose | 0.082 ± 0.06 | n.d. | 294.2 ± 9.6 | 3,587 ± 16 | |
| Neu5Ac-GM2 | 0.735 ± 0.031 | n.d. | 48.09 ± 3.84 | 65.42 ± 5.30 | |
| pNP-βNAGlu | 6.17 ± 0.92 | n.d. | 53.40 ± 2.58 | 8.65 ± 1.53 | |
| Acetylchitotetraose | 0.062 ± 0.051 | n.d. | 47.54 ± 8.16 | 766.8 ± 16 | |
| Acetylchitotriose | 9.96 ± 0.74 | n.d. | 26.71 ± 0.97 | 2.68 ± 0.13 | |
| Glycoprotein feutin2 | n.d. | 948.3 ± 18.46 | n.d. | n.d. | |
| T0216 | pNP-NAβGlu | 0.025 ± 0.006 | 3.33 ± 0.28 | 1,194 ± 2 | 47,811 ± 2,424 |
| pNP-NAβGal | 2.78 ± 1.09 | 0.28 ± 0.05 | 36.35 ± 1.51 | 15.7 ± 6.7 | |
| Glycoprotein feutin2 | n.d. | 0.14 ± 0.05 | n.d. | n.d. | |
| T0217 | pNP-NAβGlu | 0.028 ± 0.012 | 0.030 ± 0.004 | 9.88 ± 0.25 | 436.9 ± 19.2 |
| T0218 | pNP-NAβGlu | 0.134 ± 0.015 | 121.5 ± 3.59 | 83,512 ± 3,179 | 633,820 ± 946 |
| pNP-NAβGal | 0.326 ± 0.037 | 4.66 ± 0.12 | 2,191 ± 54 | 6,833 ± 96 | |
| Glycoprotein feutin2 | n.d. | 7.80 ± 0.31 | n.d. | n.d. | |
| T0220 | pNP-NAβGlu | 0.042 ± 0.004 | 0.07 ± 0.002 | 23.37 ± 0.92 | 563.6 ± 75.6 |
| pNP-NAβGal | 2.30 ± 0.17 | 0.03 ± 0.001 | 1.15 ± 0.03 | 0.5 ± 0.05 | |
| T0201 | pNP-αGal | 6.22 ± 0.93 | 10.44 ± 0.54 | 343.06 ± 10.39 | 56.67 ± 10.14 |
| T0215 | pNP-αGal | 0.062 ± 0.008 | 0.12 ± 0.008 | 79.83 ± 3.71 | 1,317 ± 230 |
| pNP-βAPyr | 3.30 ± 0.73 | 0.42 ± 0.04 | 21.53 ± 1.66 | 6.98 ± 2.05 | |
| Galactose-α-1,3-galactose | 0.048 ± 0.009 | n.d. | 98.51 ± 8.51 | 2,052 ± 95 | |
| Globotriaosylceramide | 0.096 ± 0.001 | n.d. | 72.01 ± 6.4 | 750.1 ± 64 | |
| Globotriaosylsphingosine | 0.058 ± 0.007 | n.d. | 93.45 ± 4.13 | 1,611 ± 50 | |
| T0199 | pNP-αRham | 1.08 ± 0.15 | 2.29 ± 0.11 | 1,542 ± 1308 | 1,473 ± 325 |
| T0204 | pNP-βMan | 0.127 ± 0.036 | 12.1 ± 0.95 | 1,622 ± 82 | 14,090 ± 4,637 |
| T0207 | pNP-β-Fuc | 0.0118 ± 0.003 | 1.92 ± 0.12 | 2,273 ± 31 | 206,615 ± 5,513 |
| pNP-βGal | 0.199 ± 0.032 | 1.87 ± 0.07 | 1,222 ± 33 | 6,334 ± 1,186 | |
| pNP-αAPyr | 1.732 ± 0.461 | 9.13 ± 1.64 | 602.86 ± 34.62 | 380.7 ± 121.2 | |
| T0209 | pNP-αFuc | 30.53 ± 17.64 | 5.91 ± 2.52 | 136.4 ± 5.98 | 6.88 ± 4.17 |
1
Values in Δfluorescence min−1.
2
Activity as µg sialic acid mg−1 feutin.
3
For comparisons, all reactions were performed in 50 mM Britton and Robinson buffer, pH 7.0 and 30 °C.