Elution profile of α-MMC and MAP30 on gel filtration and SP column.

(A) Chromatography of fraction from SP-Sepharose FF on Sephacryl S-100HR column. After concentration, sample was added to the Sephacryl S-100HR chromatography (2.5 cm×120 cm) and eluted with a pH 6.5 10 mM sodium phosphate buffer containing 0.15 M NaCl at flow rate of 2 ml/min. Peak I was collected fraction. (B) Chromatography of fraction from Sephacryl S-100 column on Macro-Cap-SP Column. After desalting out and exchanging buffer of fraction from Sephacryl S-100, chromatography was loaded to Macro-Cap-SP column and eluted with 10 mM NaCl in pH 6.0 50 mM sodium phosphate. Finally a linear gradient from 50 mM NaCl to 150 mM NaCl in the same buffer at flow rate of 2 ml/min was used as eluent. Two fractions were collected and identified as α-MMC and MAP30, respectively.