title: Immobilization and characterization of a new regioselective and enantioselective lipase obtained from a metagenomic library.
abstract: In previous work, a new lipase and its cognate foldase were identified and isolated from a metagenomic library constructed from soil samples contaminated with fat. This new lipase, called LipG9, is a true lipase that shows specific activities that are comparable to those of well-known industrially-used lipases with high activity against long-chain triglycerides. In the present work, LipG9 was co-expressed and co-immobilized with its foldase, on an inert hydrophobic support (Accurel MP1000). We studied the performance of this immobilized LipG9 (Im-LipG9) in organic media, in order to evaluate its potential for use in biocatalysis. Im-LipG9 showed good stability, maintaining a residual activity of more than 70% at 50 °C after incubation in n-heptane (log P 4.0) for 8 h. It was also stable in polar organic solvents such as ethanol (log P -0.23) and acetone (log P -0.31), maintaining more than 80% of its original activity after 8 h incubation at 30 °C. The synthesis of ethyl esters was tested with fatty acids of different chain lengths in n-heptane at 30 °C. The best conversions (90% in 3 h) were obtained for medium and long chain saturated fatty acids (C8, C14 and C16), with the maximum specific activity, 29 U per gram of immobilized preparation, being obtained with palmitic acid (C16). Im-LipG9 was sn-1,3-specific. In the transesterification of the alcohol (R,S)-1-phenylethanol with vinyl acetate and the hydrolysis of the analogous ester, (R,S)-1-phenylethyl acetate, Im-LipG9 showed excellent enantioselectivity for the R-isomer of both substrates (E> 200), giving an enantiomeric excess (ee) of higher than 95% for the products at 49% conversion. The results obtained in this work provide the basis for the development of applications of LipG9 in biocatalysis.
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná, Cx. P. 19046 Centro Politécnico, Curitiba 81531-980, Paraná, Brazil.
authors.forename: Robson Carlos
authors.initials: RC
authors.lastname: Alnoch
authors.affiliation: Instituto Federal do Paraná-Campus Irati, Irati 84500-000, Paraná, Brazil.
authors.forename: Viviane Paula
authors.initials: VP
authors.lastname: Martini
authors.affiliation: Agência Tecpar de Inovação, Instituto de Tecnologia do Paraná-Tecpar, Curitiba 81350-010, Paraná, Brazil.
authors.forename: Arnaldo
authors.initials: A
authors.lastname: Glogauer
authors.affiliation: Departamento de Química, Universidade Federal do Paraná, Cx. P. 19081 Centro Politécnico, Curitiba 81531-980, Paraná, Brazil.
authors.forename: Allen Carolina dos Santos
authors.initials: AC
authors.lastname: Costa
authors.affiliation: Departamento de Química, Universidade Federal do Paraná, Cx. P. 19081 Centro Politécnico, Curitiba 81531-980, Paraná, Brazil.
authors.forename: Leandro
authors.initials: L
authors.lastname: Piovan
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná, Cx. P. 19046 Centro Politécnico, Curitiba 81531-980, Paraná, Brazil.
authors.forename: Marcelo
authors.initials: M
authors.lastname: Muller-Santos
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná, Cx. P. 19046 Centro Politécnico, Curitiba 81531-980, Paraná, Brazil.
authors.forename: Emanuel Maltempi
authors.initials: EM
authors.lastname: de Souza
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná, Cx. P. 19046 Centro Politécnico, Curitiba 81531-980, Paraná, Brazil.
authors.forename: Fábio
authors.initials: F
authors.lastname: de Oliveira Pedrosa
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná, Cx. P. 19046 Centro Politécnico, Curitiba 81531-980, Paraná, Brazil.
authors.forename: David Alexander
authors.initials: DA
authors.lastname: Mitchell
authors.affiliation: Departamento de Química, Universidade Federal do Paraná, Cx. P. 19081 Centro Politécnico, Curitiba 81531-980, Paraná, Brazil.
authors.forename: Nadia
authors.initials: N
authors.lastname: Krieger
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D055162:Biocatalysis; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000235:genetics; D004952:Esters; D005227:Fatty Acids / Q000737:chemistry; D008049:Lipase / Q000737:chemistry* / Q000235:genetics; D054892:Metagenome
chemical_list: D001426:Bacterial Proteins; D004800:Enzymes, Immobilized; D004952:Esters; D005227:Fatty Acids; D008049:Lipase; C504169:lipase foldase