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Table 1 A comparison of purification efficiency between Ni-NTA resin and ITC

From: Multifunctional elastin-like polypeptide renders β-glucosidase enzyme phase transition and high stability

Proteins Total protein (mg) Specific activity (unit/mg) Total activity (unit) Yield (%) Purification (fold)
Cell lysatea (all proteins of E. coli contained EP) 3.25 ± 0.39 0.08 ± 0.004 0.263 ± 0.06   
Cell lysate with expression of BglucHb(all proteins of E. coli contained BglucH) 6.63 ± 0.53 5.75 ± 0.042 37.59 ± 0.89   
BglucHb in cell lysate 6.63 ± 0.53 5.67 ± 0.35* 37.33 ± 0.78 100.0 1.0
BglucLEHc in cell lysate (all proteins of E. coli contained BglucLEH) 5.23 ± 0.34 6.07 ± 0.46 31.32 ± 0.65   
BglucLEHc in cell lysate 5.23 ± 0.34 5.99 ± 0.54* 31.06 ± 0.63 100.0 1.0
Purified BglucH by Ni-NTA resin 0.41 ± 0.09 68.85 ± 1.19 28.23 ± 0.51 75.62 12.14
Purified BglucLEH by Ni-NTA resin 0.36 ± 0.04 68.22 ± 1.27 24.56 ± 0.59 79.07 11.39
Purified BglucLEH by one round of ITC 0.32 ± 0.05 75.50 ± 1.54 24.16 ± 0.68 77.78 12.60
Purified BglucLEH by two rounds of ITC 0.27 ± 0.03 76.22 ± 1.43 20.58 ± 0.47 66.26 12.72
  1. EP empty plasmid pET-28a(+)
  2. a,b,c2.5 mL cell extracts was obtained through ultrasonication treatment of E. coli collected from 75 mL LB culture
  3. * Indicates that the specific activity of the empty carrier has been removed