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Table 1 Purification of the recombinant Tth xynB3 β-xylosidase

From: Biochemical properties of a novel thermostable and highly xylose-tolerant β-xylosidase/α-arabinosidase from Thermotoga thermarum

Purification step

Total volume (mL)

Total activity (U)

Total protein (mg)

Specific activity (U/mg)

Recovery (%)

Purification (fold)

Crude extracta

10

2394

122

20

100

1

Heat treatmentb

10

2155

39

55

90

2.8

Ni affinity chromatographyc

1

2011

17

116

84

5.8

  1. a The recombinant strain was grown in LB medium (200 ml) with 1 μg ampicillin/ml at 37°C to OD600 0.8 and was incubated further with isopropyl-β-thiogalactopyranoside (IPTG) for 12 h. The cells were harvested by centrifugation at 10,000 g for 15 min at 4°C and resuspended in 10 ml imidazole buffer (10 mL of 5 mM imidazole, 0.5 mM NaCl, and 20 mM Tris–HCl buffer, pH 7.9), followed by sonication.
  2. b The cell extracts after sonication were heat treated at 70°C for 30 min, and then cooled in an ice bath, centrifuged at 15,000 g for 20 min at 4°C and the supernatant was kept.
  3. c The obtained supernatants were loaded on to an immobilized metal affinity column (Novagen, USA), and eluted with 0.4 M imidazole, 0.5 M NaCl, and 20 mM Tris–HCl buffer (pH 7.9).