Table 2.
Purification methods and biochemical characteristic of L-asparaginase produced by microbes.
| Strains | Purification | Characterization | References | |||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Method | SA (IU/mg) | PF | PY (%) | Opt. pH | Opt. temp (°C) |
Km (mM) | Vmax/Kcat(s−1) 1 | Mol. Wt. (kDa) (Structural Form) | ||
| Thermococcus kodakarensis | Superdex 200 10/300 GL gel column | 767 μmol/min/mg |
- | - | 7.0 | 85 | 3.1 | 833 μmol/min/mg | 70 (homodimer) | [25] |
| Pyrobaculum calidifontis | Superdex 200 10/300 GL gel filtration column | - | - | - | 6.5 | at or above 100 | 4.5 | 355 μmol/min/mg | 65 (dimer) | [43] |
| Pyrococcus yayanosii CH1 | nickel affinity column | 1483.81 | 50.62 | 60.92 | 8.0 | 95 | 6.5 | 2929 μmol/min | 72.2 (homodimer) | [44] |
| Thermococcus zilligii AN1 TziAN1_1 | nickel affinity column | 5278 | - | - | 8.5 | 90 | 6.08 | 3267 | 71 (homodimer) | [12] |
| Lactobacillus casei subsp.casei ATCC 393 | nickel affinity column | 0.419 | 9.78 | 99.8 | 6 | 40 | 0.012 | 1.576 mM/min | 35 (monomer) | [51] |
| Pseudomonas fluorescens MTCC 8127 | nickel affinity column | 26 | 4.12 | 85.4 | 7.5 | 37 | 50 | 4.032 M/min | 140 (homotetramer) | [94] |
| Lactobacillus reuteri DSM 20016 | nickel affinity column | 0.63 | 10.5 | 92 | 6 | 30 | 0.3332 | 14.06 mM/min | 35 (monomer) | [95] |
| Halomonas elongate IBRC-M10216 | nickel affinity column | 1510 | 14.5 | 27 | 6–9 | 37 | 5.6 | 2.2 μmol/min | 70–80 (homodimer) | [54] |
| Bacillus brevis | sulphopropyl Sephadex column | 9.89 | 89.9 | 15 | 6.5 | 37 | 35 | 0.77 IU | 32 (monomer) | [49] |
| Bacillus subtilis sp. KDPS-1 | ammonium sulphate precipitation and DEAE column | 97.04 | 12.11 | 84.89 | 5.0 | 37 | - | - | 97.4 - |
[50] |
| Pseudomonas otitidis | ammonium sulphate precipitation, DEAE-cellulose Column and Sephadex G-100 Column | 107.84 | 151.88 | 38.9 | 7.5 | 40 | - | - | 205 (homohexamer) | [96] |
| Aquabacterium sp. A7-Y (abASNase2) | nickel affinity column | 458.9 | - | - | 9.0 | 60 | 1.8 | 241.9 | 33 (monomer) | [20] |
| Penicillium crustosum NMKA 511 | ammonium sulphate precipitation, DEAE-Sephadex column and Sephadex G-100 column | 9.84 | 6.47 | 36.3 | 6.67 | 36.9 | 3.79 | 499.8 µmol/min/mg | 41.3 and 44.4 (heterodimer) |
[19] |
| Aspergillus oryzae CCT 3940 | ammonium sulphate precipitation, Q Sepharose™ column, SP Sepharose™ column and CM Sepharose™ column | 282 | 28.6 | 6 | 5–8 | 40–50 | 2.10 | 35.8 U/mL | 115 - |
[37,67] |
| Saccharomyces cerevisiae | nickel affinity column and PD-10 Desalting column | - | - | - | 8.6 | 40 | 0.075 2 | 0.042 μmol/min | 41.4 - |
[73] |
| Candida utilis ATCC9950 | acetone precipitation and Q-sepharose column | 7853 | 10.02 | 82.7 | - | - | - | - | 40 - |
[97] |
| Spirulina maxima | ammonium sulphate precipitation and Sephadex G-200 column | 19.1 | 10.91 | 86.45 | 8.5 | 37 | - | - | - - |
[74] |
SA, specific activity; PY, protein yield; PF, purification fold; Opt. pH, optimum pH; Opt. temp., optimum temperature; Mol. Wt., molecular weight; Km, Michaelis constant. 1: Vmax was replaced with Kcat in some references and Vmax employed different unit in different references. 2: L-asparaginase 1 from Saccharomyces cerevisiae has allosteric behavior similar to type I enzymes, but with a low K 0.5 = 75 μM as for therapeutic type II.