Produção Nacional
Revisado por pares
Modulation of a lipase from Staphylococcus warneri EX17 using immobilization techniques
2009; Elsevier BV; Volume: 60; Issue: 3-4 Linguagem: Inglês
10.1016/j.molcatb.2009.04.007
ISSN1873-3158
AutoresGiandra Volpato, Marco Filice, Rafael C. Rodrigues, Júlio Xandro Heck, José M. Guisán, César Mateo, Marco Antônio Záchia Ayub,
Tópico(s)Electrochemical sensors and biosensors
ResumoThis research describes the immobilization on glyoxyl, cyanogen bromide or octyl agarose beads of a purified lipase from Staphylococcus warneri strain EX17 (SWL), and the effect on its properties. The immobilization on glyoxyl-agarose at pH 10 and 25 °C, conditions in which the enzyme is readily inactivated, required the stabilization of the soluble enzyme. This was attained by the addition of 25% glycerol. Using this additive, immobilization on glyoxyl-agarose beads proceeded very quickly with good activity retention around 80%. This was the most stable preparation under thermal inactivation at pH 5, 7 and 9, in the presence of either cosolvents or detergents. This preparation was hyperactivated by concentrations of Triton X-100, which would produce negative effects over enzyme activity when using the other SWL preparations. Immobilized SWL preparations hydrolyzed different chiral esters, such as (±)-methyl mandelate, (±)-2-O-butyryl-2-phenylacetic acid, and (±)-2-hydroxy-4-phenyl-butyric acid ethyl ester, being its specificity depended on the immobilization protocol. The enantiospecificity was also strongly modulated by the immobilization. Thus, using HPBEt as substrate, octyl-SWL exhibited an opposite enantiospecificity to the other two biocatalysts. This preparation was the most enantioselective in the hydrolysis of (±)-2-O-butyryl-2-phenylacetic acid (E = 56.3).
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