Activity and stability of chemically modified Candida antarctica lipase B adsorbed on solid supports

• Published in: Applied microbiology and biotechnology 1999-11, Vol.52 (6), p.791-796
• Main Authors: KOOPS, B. C, PAPADIMOU, E, VERHEIJ, H. M, SLOTBOOM, A. J, EGMOND, M. R
• Format: Article
• Language: English
• Subjects: Absorption; Adsorption; Biological and medical sciences; Biotechnology; Candida - enzymology; Candida antarctica; Cation Exchange Resins - metabolism; Dextrans - pharmacology; Enzymes, Immobilized - drug effects; Enzymes, Immobilized - metabolism; Fundamental and applied biological sciences. Psychology; Fungal Proteins - drug effects; Fungal Proteins - metabolism; Immobilization of enzymes and other molecules; Immobilization techniques; Indigenous species; Lipase - drug effects; Lipase - metabolism; Methods. Procedures. Technologies; Octanols - pharmacology; Polyethylene glycol; Polyethylene Glycols - metabolism; Polyethylene Glycols - pharmacology; Protein Folding; Temperature; Time Factors; Xylene
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s002530051593

The effect of various covalent chemical modifications on the transesterification activity and stability of adsorbed lipase B from Candida antarctica (CALB) was studied in 2-butanone and o-xylene. CALB species modified with either polyethylene glycol 2000 monomethyl ether (MPEG), polyethylene glycol 300 mono-octyl ether (OPEG) or n-octanol (OCT) were used in combination with a hydrophobic (Accurel) and a hydrophilic (Duolite) support. The thermostabilities of adsorbed CALB in both solvents, and that of free CALB in o-xylene were not influenced by the modifications. In contrast, the thermostability of free CALB in 2-butanone decreased 2.5-fold after MPEG modification and increased 1.5-fold after modification with OPEG and n-octanol, compared to that of native CALB. The activities of the native and modified CALB species were up to 9-fold higher after adsorption onto Accurel than those of the corresponding free enzymes. Adsorption of these enzyme species onto Duolite only resulted in a 2- to 3-fold increase in the activity of OPEG- and OCT-modified CALB. The modified CALB species adsorbed onto Accurel show similar or up to 2-fold lower activities than do native adsorbed CALB species, while 1.5- to 6-fold higher activities were found for modified CALB species adsorbed onto Duolite. We propose that hydrophobic modifiers induce conformational changes of CALB during adsorption on a hydrophobic support whereas all three modifiers protect CALB from structural alterations during adsorption onto a hydrophilic support.