Hydrophobic interaction chromatography of Chromobacterium viscosum lipase on polypropylene glycol immobilised on Sepharose.
The fractionation of Chromobacterium viscosum lipase was performed using a polypropylene glycol-Sepharose gel. The influence of mobile phase composition on the adsorption of lipase on the gel was studied and it was found that the retention of lipase depends on the salt used and increased with increasing the ionic strength. The retention was not strongly affected by changing the pH value of the mobile phase. By using 20% (w/v) ammonium sulphate in phosphate buffer a total retention of lipase on the column was obtained and by simply decreasing the ionic strength of the buffer, desorption of lipase could be achieved. The chromatographic purification of Chromobacterium viscosum lipase by hydrophobic interaction chromatography on Sepharose CL-6B modified by covalent immobilisation of 1,4-butanediol diglycidyl ether, polyethylene glycol and polypropylene glycol was also compared.
Diogo
M M
MM
Departamento de Química, Universidade da Beira Interior, Covilhã, Portugal.
Silva
S
S
Cabral
J M
JM
Queiroz
J A
JA
D002072:Butylene Glycols / Q000737:chemistry; D002847:Chromatography, Agarose / Q000379:methods*; D002861:Chromobacterium / Q000201:enzymology*; D008049:Lipase / Q000302:isolation & purification*; D011092:Polyethylene Glycols / Q000737:chemistry; D011108:Polymers / Q000737:chemistry*; D011409:Propylene Glycols / Q000737:chemistry*
D002072:Butylene Glycols; D011108:Polymers; D011409:Propylene Glycols; C012504:polypropylene glycol; D011092:Polyethylene Glycols; C014376:1,4-bis(2,3-epoxypropoxy)butane; D008049:Lipase