Different covalent immobilizations modulate lipase activities of Hypocrea pseudokoningii

Autor: Benevides C. C. Pessela, Fernanda Dell Antonio Facchini, Marita Gimenez Pereira, Paulo Ricardo Heinen, Ana Claudia Vici, Susana Velasco-Lozano, Aline M. Polizeli, Jose M. Guisan, Maria de Lourdes Teixeira de Moraes Polizeli, Sonia Moreno-Pérez, Mariana Cereia, Gloria Fernández-Lorente, João Atílio Jorge
Přispěvatelé: Fundações de Amparo à Pesquisa (Brasil), Fundação de Amparo à Pesquisa do Estado de São Paulo, Conselho Nacional de Desenvolvimento Científico e Tecnológico (Brasil), Guisán, José Manuel [0000-0003-1627-6522], Polizeli, Maria de Lourdes T. M. [0000-0002-5026-6363], Guisán, José Manuel, Polizeli, Maria de Lourdes T. M.
Rok vydání: 2017
Předmět:
0106 biological sciences
Protein Denaturation
Pharmaceutical Science
01 natural sciences
Analytical Chemistry
Substrate Specificity
chemistry.chemical_compound
Drug Discovery
Enzyme Stability
Organic chemistry
Cyanogen Bromide
biology
Protein Stability
Hydrolysis
Sepharose
Hypocrea pseudokoningii
Temperature
Stereoisomerism
Hydrogen-Ion Concentration
Hydrolysis of oils
Cross-Linking Reagents
Eicosapentaenoic Acid
Chemistry (miscellaneous)
Molecular Medicine
Stability
SOLVENTE
Immobilized enzyme
Docosahexaenoic Acids
Hypocrea
hydrolysis of oils
Lipase activity modulation
enzyme immobilization
lipase activity modulation
stability
Article
lcsh:QD241-441
lcsh:Organic chemistry
010608 biotechnology
Humans
Enzyme immobilization
Physical and Theoretical Chemistry
Lipase
Ethanol
Chromatography
010405 organic chemistry
Organic Chemistry
Enzymes
Immobilized
0104 chemical sciences
Enzyme Activation
chemistry
Glutaral
biology.protein
Solvents
Racemic mixture
Glutaraldehyde
Methanol
Enantiomer
Oils
Zdroj: Digital.CSIC. Repositorio Institucional del CSIC
instname
Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry
Molecules; Volume 22; Issue 9; Pages: 1448
Molecules, Vol 22, Iss 9, p 1448 (2017)
Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual)
Universidade de São Paulo (USP)
instacron:USP
Popis: This article belongs to the Special Issue Lipases and Lipases Modification.
Enzyme immobilization can promote several advantages for their industrial application. In this work, a lipase from Hypocrea pseudokoningii was efficiently linked to four chemical supports: agarose activated with cyanogen bromide (CNBr), glyoxyl-agarose (GX), MANAE-agarose activated with glutaraldehyde (GA) and GA-crosslinked with glutaraldehyde. Results showed a more stable lipase with both the GA-crosslinked and GA derivatives, compared to the control (CNBr), at 50 °C, 60 °C and 70 °C. Moreover, all derivatives were stabilized when incubated with organic solvents at 50%, such as ethanol, methanol, n-propanol and cyclohexane. Furthermore, lipase was highly activated (4-fold) in the presence of cyclohexane. GA-crosslinked and GA derivatives were more stable than the CNBr one in the presence of organic solvents. All derivatives were able to hydrolyze sardine, açaí (Euterpe oleracea), cotton seed and grape seed oils. However, during the hydrolysis of sardine oil, GX derivative showed to be 2.3-fold more selectivity (eicosapentaenoic acid (EPA)/docosahexaenoic acid (DHA) ratio) than the control. Additionally, the types of immobilization interfered with the lipase enantiomeric preference. Unlike the control, the other three derivatives preferably hydrolyzed the R-isomer of 2-hydroxy-4-phenylbutanoic acid ethyl ester and the S-isomer of 1-phenylethanol acetate racemic mixtures. On the other hand, GX and CNBr derivatives preferably hydrolyzed the S-isomer of butyryl-2-phenylacetic acid racemic mixture while the GA and GA-crosslink derivatives preferably hydrolyzed the R-isomer. However, all derivatives, including the control, preferably hydrolyzed the methyl mandelate S-isomer. Moreover, the derivatives could be used for eight consecutive cycles retaining more than 50% of their residual activity. This work shows the importance of immobilization as a tool to increase the lipase stability to temperature and organic solvents, thus enabling the possibility of their application at large scale processes.
This work was supported by grants from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, process No. 2013/50892-5; 2010/52322-3), Conselho de Desenvolvimento Científico e Tecnológico (CNPq process No. 406838/2013-5; 563260/2010-6). This project is also part of the National Institute of Science and Technology of the Bioethanol (No. 574002/2008-1), CNPq—Ciência sem Fronteira (No. 242775/2012-8). JAJ and MLTMP are Research Fellows of CNPq. MGP and ACV are supported by CNPq. FDAF was recipient of a FAPESP Fellowship.
Databáze: OpenAIRE
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