Immobilization of Candida rugosa lipase on magnetized Dacron: kinetic study.
Candida rugosa lipase has been covalently immobilized on ferromagnetic azide polyethyleneterepthalate (Dacron) with specific activity retention of 16% for 4-nitrophenyl palmitate and 24% for hydrolysis of triolein in hexane. The immobilized enzyme was more thermal stable than the soluble one, retaining 78.8% of the activity after 1 h at 60 degrees C. Also, this immobilized derivative was stable at the storage at 4 degrees C. It has been used 5 cycles for pNPP hydrolysis without loss of activity. Soluble and immobilized Candida rugosa lipase showed a Michaelian behavior for fatty acid 4-nitrophenyl esters and different apparent K(M) values: 0.110 mM and 0.124 mM (4-nitrophenyl palmitate - C16); 0.193 mM and 0.235 mM (4-nitrophenyl laurate - C12) and 0.206 mM and 0.119 mM (4-nitrophenyl butyrate - C4), respectively. The immobilized lipase was more efficient for catalyzing the hydrolysis of 4-nitrophenyl esters with short chain length fatty acid (4-NPB - C4) than soluble enzyme. The ferromagnetic Dacron-lipase derivative was able to catalyze the synthesis of triolein from glycerol and oleic acid with 50% of conversion after 72 h at 40 degrees C.
Pimentel
M C B
MC
Dept. Bioquímica (UFPE), Lab. Immunopatologia Keizo Asami, Federal University of Pernambuco, Recife, Brazil.
Leāo
A B F
AB
Melo
E H M
EH
Ledingham
W M
WM
Filho
J L Lima
JL
Sivewright
M
M
Kennedy
J F
JF
D019840:2-Propanol; D001709:Biotechnology / Q000379:methods*; D002175:Candida / Q000201:enzymology*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D004952:Esters / Q000378:metabolism; D006358:Hot Temperature; D007700:Kinetics; D008049:Lipase / Q000378:metabolism*; D008280:Magnetics; D011093:Polyethylene Terephthalates; D012997:Solvents; D013379:Substrate Specificity
D004800:Enzymes, Immobilized; D004952:Esters; D011093:Polyethylene Terephthalates; D012997:Solvents; D008049:Lipase; D019840:2-Propanol