Stability and catalytic properties of lipase immobilized on chitosan encapsulated magnetic nanoparticles cross-linked with genipin and glutaraldehyde

Abstract

BACKGROUND Among many cross‐linkers for enzyme immobilization, genipin (Gen) and glutaraldehyde (Glu) as cross‐linkers have been commonly investigated to improve the catalytic performance of enzyme immobilized on chitosan encapsulated magnetic nanoparticles (CMNPs). In this work, we compared the stability and catalytic properties of lipase Candida rugosa (CRL) immobilized on CMNPs cross‐linked with Gen and Glu, respectively. The CMNPs were first characterized by TEM, XRD and FT‐IR. Moreover, some crucial parameters affecting catalytic performances were optimized for Gen‐CMNPs‐CRL and Glu‐CMNPs‐CRL preparation. RESULTS The Gen‐CMNPs‐CRL showed maximum activity at pH 8.0 and 40°C, and retained more than 95% of its initial activity after 7 days storage at 25°C. After 5 cycles re‐usage, Gen‐CMNPs‐CRL still retained over 80% of its initial activity, while Glu‐CMNPs‐CRL retained only 26% of its initial activity. Kinetic studies confirmed that Gen‐CMNPs‐CRL and Glu‐CMNPs‐CRL presented higher substrate affinity characteristics (Km) than free CRL. FT‐IR analysis showed that the variance of β‐sheet element in the secondary structure of CRL might contribute to the stability and activity enhancement of Gen‐CMNPs‐CRL. CONCLUSIONS Gen‐CMNPs‐CRL showed higher pH, temperature, storage and operational stabilities than Glu‐CMNPs‐CRL. Thus, genepin is a better promising cross‐linker than glutaraldehyde for lipase immobilization on CMNPs. © 2015 Society of Chemical Industry

Co-Immobilization of Enzymes and Magnetic Nanoparticles by Metal-Nucleotide Hydrogelnanofibers for Improving Stability and Recycling

“…What’s more, enzymes immobilized on solid supports show improved essential properties in actual application. Various materials have been used for immobilizing enzymes [ 13 ], such as carboxyl-functionalized graphene oxide [ 14 ], polyurethane foam support [ 15 ], SBA-15 [ 16 ], and chitosan [ 17 ]. Nowadays, the exploration of novel support materials and technologies are still attracting much attention.…”

“…Lipases are prominent biocatalysts to catalyze reactions in diversified areas [ 44 ]. Candida rugosa lipase (CRL) is a versatile enzyme to convert various substrates into useful products [ 17 ]. Herein, we reported a convenient, efficient, and high capacity magnetic immobilization method for enzymes by the co-entrapment of Fe 3 O 4 nanoparticles and CRL simultaneously within Zn/AMP nanofiber supports ( Scheme 1 ).…”

Genipin as An Emergent Tool in the Design of Biocatalysts: Mechanism of Reaction and Applications

“…Genipin permitted to prepare the biocatalysts with the best performance; for example, after five reaction cycles, the biocatalyst prepared using glutaraldehyde exhibited only 26% of its initial activity while the one prepared using genipin maintained over 80%. In fact, this biocatalyst was more stable under drastic pH and temperature, and also presented better operational stabilities than the one prepared using glutaraldehyde [100].…”

“…This biocatalyst showed enhanced stability (at drastic pH values or high temperatures) and retained more than 60% of its initial value after five hydrolytic cycles [99]. In another paper, lipase from Candida rugosa was immobilized on chitosan encapsulated magnetic nanoparticles using glutaraldehyde or genipin [100]. Genipin permitted to prepare the biocatalysts with the best performance; for example, after five reaction cycles, the biocatalyst prepared using glutaraldehyde exhibited only 26% of its initial activity while the one prepared using genipin maintained over 80%.…”

Optical monitoring of adipose tissue destruction under encapsulated lipase action

“…Such materials as mesoporous silica , microemulsion‐based organogels , sol‐gel matrix , chitosan, encapsulated magnetic nanoparticles , polycaprolactone nanofibers , have been previously applied for lipase encapsulation. However, there are some limitations, which are inherent to enzyme encapsulation, including poor loading efficiency, instability, or even enzyme leakage.…”