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Table 1 Overview on the protein concentration (mg/ml) and total amount of protein (mg) of each step along the purification process of LsAA9A and NcAA9C expressed with either the pBSY3Z or the pBSYPGCW14Z expression plasmid and the native LPMO SP, from the crude supernatant to the pure protein

From: Novel molecular biological tools for the efficient expression of fungal lytic polysaccharide monooxygenases in Pichia pastoris

Enzyme

Plasmid + SP

Supernatant

Fivefold concentrated

Purified (obtained from 50 ml cultivation supernatant)

Recovered protein

  

mg

mg/ml

mg

mg/ml

mg

mg/ml

%

LsAA9A

pBSY3Z + native

21 ± 5

0.042 ± 0.010

22.5 ± 0.4

0.225 ± 0.004

0.61 ± 0.01

1.2 ± 0.02

27

pBSYPGCW14Z + native

28 ± 2

0.056 ± 0.004

24.7 ± 1.7

0.247 ± 0.017

0.6 ± 0.03

1.4 ± 0.05

24

NcAA9C

pBSY3Z + native

22.5 ± 1.5

0.045 ± 0.003

20.7 ± 2.3

0.207 ± 0.023

0.39 ± 0.02

0.8 ± 0.05

18

pBSYPGCW14Z + native

76 ± 7

0.153 ± 0.014

70.5 ± 0.5

0.705 ± 0.005

2.08 ± 0.06

2.9 ± 0.03

29

  1. For precise measurements, the volume of the supernatant and the fivefold concentrate was adjusted to 500 ml and 100 ml, respectively. Please note that the values reported in the column labeled with “purified” were obtained from 50 ml cultivation supernatant