Fig. 4
Large-scale purification of R. marinus CelA expressed and secreted in S. lividans culture supernatants. Protein samples (15 µg/lane) from the various purification steps (see “Methods”) were analyzed as in Fig. 2a and were stained with Coomassie blue. Lane 1: mass markers as in Fig. 1. Lane 2: culture supernatants from S. lividans TK24 with pIJ486 expressing SPVsi-CelA grown in NB for 48 h; lane 3: the culture supernatant after concentration with rotary evaporator; lane 4: The 35–55% ammonium sulfate fraction; lane 5: the elution fraction with the highest CelA activity from Q-Sepharose; lane 6: the fraction from lane 5 after heating at 80 °C for 4 h and centrifugal removal of precipitates