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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