Figure 1 of Chitpinityol, Mol Vis 4:1, 1998.

Figure 1. Refolding and activation of prochymosin and chymosin.

The (*) indicates the step of the refolding process where [alpha]-crystallin was added, after dilution with phosphate buffer. Steps 1 through 7 are the refolding process. See text for details.

Protein inclusion bodies

(<1 K)

Solubilized in 10 w/v 8 M urea buffer, 25 °C, 1 h

(<1 K)

Centrifuged

(<1 K)

Supernatant of 8 M urea mixture

(<1 K)

(1) Slowly diluted in 11.5 v/v of phosphate buffer pH 10.7(*), 25 °C, 1 h

(<1 K)

(2) Adjusted pH to 8, 25 °C, 1 h

(<1 K)

(3) Dialyzed in buffer B, 4 °C, overnight

(<1 K)

(4) Centrifuged, discarded pellets

(<1 K)

Folded prochymosin

(<1 K)

Activated prochymosin to chymosin

(<1 K)

(5) Adjusted to pH 2, 25 °C, 2 h

(<1 K)

(6) Adjusted to pH 6.3, 25 °C, 1 h

(<1 K)

(7) Centrifuged 13000 g, 20 min

(<1 K)

Active chymosin

Chitpinityol, Mol Vis 1998; 4:1<http://www.emory.edu/molvis/v4/p1>
©1998 Molecular Vision
ISSN 1090-0535