Serum Amo V max Km Kip Rep Cap
  s-1 x M-1 pmol/min/mg μM by biotin (μM) s-1 x M x 10-3 s--1
Healthy control (without inflammation)
Healthy adult control  9 (20 y, male)
Fresh 19.1 123 8.16 689 107 45.2
Heat treatment 16.0 105 8.30 861 114 42.7
With fucoidan 14.2 95.2 8.47 794 95.5 36.8
With neuraminidase 5.90 76.9 16.5 628 61.1 19.0
With fucosidase 10.5 120 14.5 963 147 39.3
Healthy child control 4 (8 mo, female)  (gastritis)
Fresh 22.2 156 8.91 494 97.6 46.5
Heat treatment 26.2 185 8.93 506 118 55.6
With fucoidan 33.6 211 7.94 471 126 65.1
With heparin 35.2 217 7.81 346 95.1 57.9
With neuraminidase 24.0 179 9.43 247 55.9 36.6
With fucosidase 38.0 131 4.37 157 26.1 31.5
Patients            
Child patient 1 with GSD1b (3 y, female)
Before biotin            
Fresh 19.9 132 8.38 404 67.4 36.6
Heat treatment 7.95 147 23.4 478 88.9 26.6
With fucoidan 21.5 98.0 5.78 1410 175 61.3
With heparin 13.0 139 13.5 202 35.5 21.5
With neuraminidase 10.4 69.0 8.40 1140 99.6 32.2
With fucosidase 12.0 127 13.4 430 69.2 28.8
After biotin (32 w)            
Fresh 17.5 101 7.30 530 67.7 34.4
Heat treatment 9.41 113 15.2 549 78.5 27.2
With fucoidan 33.8 122 4.57 317 49.0 40.7
With heparin 21.5 99.5 5.85 769 96.7 45.6
With neuraminidase 13.9 75.8 6.90 255 24.5 18.5
With fucosidase 5.82 95.7 20.8 1250 152 29.7
Child patient 2 (3 mo, male)            
Before biotin            
Fresh 25.5 182 9.02 365 83.9 46.3
Heat treatment 8.63 122 17.9 271 41.8 19.0
With fucoidan 33.6 143 5.39 760 138 68.1
With heparin 26.3 233 11.2 765 225 76.9
With neuraminidase 9.25 44.2 6.05 296 16.6 12.4
With fucosidase 7.75 73.5 12.0 401 37.3 17.0
After biotin (1 w)            
Fresh 23.9 103 5.46 515 67.2 40.1
Heat treatment 17.7 145 10.4 661 122 46.5
With fucoidan 30.9 143 5.85 439 79.4 49.5
With heparin 25.4 154 7.69 874 171 65.9
With neuraminidase 3.22 57.5 22.6 1050 76.6 15.7
With fucosidase 5.45 78.4 18.2 1450 144 28.0
Adult patient 5 (32 y, adult female) (gait disorder, contraction (narrowing of visual field) )
Before biotin            
Fresh 35.9 112 3.95 331 46.9 41.0
Heat treatment 10.1 124 15.6 1310 206 45.6
With fucoidan 16.8 116 8.73 424 62.2 32.3
With heparin 10.5 122 14.7 1020 158 40.7
With neuramindase 13.4 79.5 7.52 549 55.3 28.3
With fucosidase 12.4 73.7 7.52 230 21.5 20.6
After biotin (1 w)            
Fresh 26.0 177 8.62 408 91.4 48.8
Heat treatment 20.1 135 8.48 737 124 49.6
With fucoidan 22.1 122 6.99 430 66.4 38.3
With heparin 12.0 79.4 8.40 1690 171 45.3
With neuraminidase 13.4 79.5 7.52 594 59.8 28.3
With fucosidase 19.7 73.7 4.74 230 21.5 20.6
Before Ebios            
Fresh 18.4 63.6 4.45 342 27.7 22.4
Heat treatment 16.6 500 38.2 265 168 52.8
With fucoidan 17.3 125 9.13 609 96.2 40.8
With heparin 21.8 155 9.01 277 54.4 34.4
With neuraminidase 7.00 120 21.7 848 129 30.0
With fucosidase 5.19 47.6 11.6 754 45.4 15.4
Before Ebios            
Fresh 18.4 63.6 4.45 342 27.7 22.4
Heat treatment 16.6 500 38.2 265 168 52.8
With fucoidan 17.3 125 9.13 609 96.2 40.8
With heparin 21.8 155 9.01 277 54.4 34.4
With neuraminidase 7.00 120 21.7 848 129 30.0
With fucosidase 5.19 47.6 11.6 754 45.4 15.4
After Ebios +Yakult (16 w)            
Fresh 24.6 141 7.25 401 71.5 41.9
Heat treatment 8.52 90.9 13.5 2760 318 52.1
With fucoidan 8.86 67.6 9.66 2430 200 42.1
With heparin 10.0 65.4 8.26 5400 446 66.8
With neuraminidase 8.76 66.2 9.57 1100 92.5 28.5
With fucosidase 10.2 155 19.2 899 176 42.4
After Ebios +Yakult (1.5 y)            
Fresh 14.2 132 11.8 417 69.9 31.5
Heat treatment 4.51 75.8 21.3 543 52.2 15.3
With fucoidan 17.7 77.2 5.51 444 43.3 27.7
With heparin 15.5 147 12.0 670 125 44.0
With neuraminidase 7.72 55.2 9.05 862 60.3 21.6
With fucosidase 12.4 90.9 9.26 271 31.1 19.6
*Parameters of biotinidase kinetics were expressed according to the ref. 9. Thermal instability test was performed as in ref. 18. Serum of a healthy adult male (Healthy adult control 9; 20 y, male) was obtained from the Okayama University. Other conditions were as described in the Materials and Methods section.
Table 3: Kinetic parameters of the glycoprotein enzyme of biotinidase: effects of glycosidases and fucoidan in vitro.*