Table 1. Details of Exoglycosidase Enzymes and Digestionsa.
| enzyme | full name | specificity | supplier | cat no. | units (U/mL) | volume (μL) |
|---|---|---|---|---|---|---|
| ABS | arthrobacter ureafaciens sialidase | α(2-3, -6, -8) linked sialic acid | NEB | P0722 | 20,000 | 1 |
| NAN-1 | recombinant sialidase | α(2-3) linked sialic acid | NEB | P0743 | 50,000 | 1 |
| BTG | bovine testes β-galactosidase | β(1-3, -4) linked galactose | Prozyme | GKX-5013 | 5 | 2 |
| SPG | Streptococcus pneumoniae β-galactosidase | β(1–4) linked galactose | Prozyme | GKX-5014 | 2 | 2 |
| CBG | coffee bean α-galactosidase | α(1–3, -4) linked galactose | Prozyme | GKX-5007 | 5 | 2.5 |
| AMF | almond meal α-fucosidase | α(1–3) linked fucose (outer-arm fucose) | NEB | P0769 | 4,000 | 1 |
| BKF | Bovine kidney α-fucosidase | α(1–6) linked fucose (core fucose), some α(1-2) outer-arm fucose | NEB | P0749 | 2,000 | 1 |
| GUH | Streptococcus pneumonia hexosamindase | β-linked GlcNAc | Prozyme | GK800050 | 40 | 1 |
| JBH | Jack bean β-N-acetylhexoaminidase | β-linked GlcNAc and GalNAc residues | Prozyme | GKX-5003 | 50 | 2 |
| JBM | Jack bean mannosidase | α(1-2, -6, -3) linked mannose | NEB | P0768 | 2,000 | 2 |
| enzyme combinations | ||||||
| 1 | UND, Undigested, no enzyme | |||||
| 2 | NAN-1 | |||||
| 3 | ABS | |||||
| 4 | ABS + BTG | |||||
| 5 | ABS + BTG + BKF | |||||
| 6 | ABS + BTG + AMF | |||||
| 7 | ABS + AMF | |||||
| 8 | ABS + BTG + BKF + AMF | |||||
| 9 | ABS + BTG + BKF + AMF + GUH | |||||
| 10 | ABS + BTG + BKF + AMF + GUH + CBG | |||||
| 11 | ABS + BTG + BKF + AMF + GUH + JBM | |||||
| 12 | ABS + SPG | |||||
| 13 | ABS + JBH | |||||
a
Digestions were performed in a total of 10 μL and in the presence of 0.05 M sodium acetate buffer (1 μL). Digestion with JBM was performed independently, in the presence of Zn2+ (0.1%), following ABS + BTG + BKF + GUH digestion on the previous day. NEB, New England Biolabs.