title: Heterofunctional hydrophilic-hydrophobic porous silica as support for multipoint covalent immobilization of lipases: application to lactulose palmitate synthesis.
abstract: Lipase-catalyzed synthesis of sugar esters, as lactulose palmitate, requires harsh conditions, making it necessary to immobilize the enzyme. Therefore, a study was conducted to evaluate the effect of different chemical surfaces of hierarchical meso-macroporous silica in the immobilization of two lipases from Pseudomonas stutzeri (PsL) and Alcaligenes sp. (AsL), which exhibit esterase activity. Porosity and chemical surface of silica supports, before and after functionalization and after immobilization, were characterized by gas adsorption and Fourier transform infrared (FTIR) spectroscopy. PsL and AsL were immobilized in octyl (OS), glyoxyl (GS), and octyl-glyoxyl silica (OGS). Hydrolytic activity, thermal and solvent stability, and sugar ester synthesis were evaluated with those catalysts. The best support in terms of expressed activity was OS in the case of PsL (100 IU g(-1)), while OS and OGS were the best for AsL with quite similar expressed activities (60 and 58 IU g(-1), respectively). At 60 °C in aqueous media the more stable biocatalysts were GS-PsL and OGS-AsL (half-lives of 566 and 248 h, respectively), showing the advantage of a heterofunctional support in the latter case. Lactulose palmitate synthesis was carried out in acetone medium (with 4% of equilibrium moisture) at 40 °C obtaining palmitic acid conversions higher than 20% for all biocatalysts, being the highest of those obtained with OGS-AsL and OS-PsL. Therefore, screening of different chemical surfaces on porous silica used as supports for lipase immobilization allowed obtaining active and stable biocatalyst to be employed in the novel synthesis of lactulose palmitate.
authors.affiliation: Escuela de Ingeniería Bioquímica, Pontificia Universidad Católica de Valparaíso , Avenida Brasil 2147, Valparaíso, Chile.
authors.forename: Claudia
authors.initials: C
authors.lastname: Bernal
authors.forename: Andres
authors.initials: A
authors.lastname: Illanes
authors.forename: Lorena
authors.initials: L
authors.lastname: Wilson
mesh_terms: D000421:Alcaligenes / Q000201:enzymology; D055162:Biocatalysis; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D057927:Hydrophobic and Hydrophilic Interactions; D007792:Lactulose / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000737:chemistry; D008049:Lipase / Q000737:chemistry / Q000378:metabolism*; D010168:Palmitates / Q000737:chemistry / Q000378:metabolism*; D010316:Particle Size; D016062:Porosity; D044223:Pseudomonas stutzeri / Q000201:enzymology; D012822:Silicon Dioxide / Q000737:chemistry / Q000378:metabolism*; D013499:Surface Properties
chemical_list: D004800:Enzymes, Immobilized; D010168:Palmitates; D007792:Lactulose; D012822:Silicon Dioxide; D008049:Lipase