title: Separation and immobilization of lipase from Penicillium simplicissimum by selective adsorption on hydrophobic supports.
abstract: Lipases are an enzyme class of a great importance as biocatalysts applied to organic chemistry. However, it is still necessary to search for new enzymes with special characteristics such as good stability towards high temperatures, organic solvents, and high stereoselectivity presence. The present work's aim was to immobilize the lipases pool produced by Penicillium simplissicimum, a filamentous fungi strain isolated from Brazilian babassu cake residue. P. simplissicimum lipases were separated into three different fractions using selective adsorption method on different hydrophobic supports (butyl-, phenyl-, and octyl-agarose) at low ionic strength. After immobilization, it was observed that these fractions' hyperactivation is in the range of 131% to 1133%. This phenomenon probably occurs due to enzyme open form stabilization when immobilized onto hydrophobic supports. Those fractions showed different thermal stability, specificity, and enantioselectivity towards some substrates. Enantiomeric ratio for the hydrolysis of (R,S) 2-O-butyryl-2-phenylacetic acid ranged from 1 to 7.9 for different immobilized P. simplissicimum lipase fractions. Asymmetry factor for diethyl 2-phenylmalonate hydrolysis ranged from 11.8 to 16.4 according to the immobilized P. simplissicimum lipase fractions. Those results showed that sequential adsorption methodology was an efficient strategy to obtain new biocatalysts with different enantioselectivity degrees, thermostability, and specificity prepared with a crude extract produced by a simple and low-cost technology.
authors.affiliation: Instituto de Química, Centro de Tecnologia, Universidade Federal do Rio de Janeiro, lab. 549-1, Ilha do Fundão, CEP 21945-970 Rio de Janeiro, RJ, Brazil.
authors.forename: Aline G
authors.initials: AG
authors.lastname: Cunha
authors.forename: Gloria
authors.initials: G
authors.lastname: Fernández-Lorente
authors.forename: Melissa L E
authors.initials: ML
authors.lastname: Gutarra
authors.forename: Juliana V
authors.initials: JV
authors.lastname: Bevilaqua
authors.forename: Rodrigo V
authors.initials: RV
authors.lastname: Almeida
authors.forename: Lúcia M C
authors.initials: LM
authors.lastname: Paiva
authors.forename: Roberto
authors.initials: R
authors.lastname: Fernández-Lafuente
authors.forename: Jose M
authors.initials: JM
authors.lastname: Guisán
authors.forename: Denise M G
authors.initials: DM
authors.lastname: Freire
mesh_terms: D000327:Adsorption; D004800:Enzymes, Immobilized / Q000378:metabolism*; D057927:Hydrophobic and Hydrophilic Interactions; D008049:Lipase / Q000302:isolation & purification* / Q000378:metabolism*; D010407:Penicillium / Q000378:metabolism*; D013237:Stereoisomerism
chemical_list: D004800:Enzymes, Immobilized; D008049:Lipase