Enhancing the thermal stability of lipases through mutagenesis and immobilization on zeolites

Abstract

The hydrolysis reaction of p-nitrophenyl butyrate catalyzed by lipases was followed with in situ UV/vis diode array spectrophotometry. Five enzymes - Candida antarctica lipase B and Fusarium solani pisi cutinase wild-type and three single-mutation variants - were tested as catalysts in homogeneous conditions and immobilized on zeolite NaY, on a polyacrylate support and as cross-linked aggregates. Using deconvolution techniques and kinetic modeling, the thermal stability of the different biocatalysts was compared in operational conditions and the results were supported by steady-state enzyme fluorescence measurements. We concluded that both the mutagenesis and the immobilization on zeolite NaY had a positive effect on the thermal stability of F. solani pisi cutinase.

Synergistic effect in the catalytic activity of lipase Rhizomucor miehei immobilized on zeolites for the production of biodiesel

“…Most of zeolites properties are related to the possibility of generating and regulating their acidbase, hydrophobic-hydrophilic character and also their strong selective adsorption affinities. Due to these properties, there are several studies reporting the use of zeolites in their pure phase not only as heterogeneous catalysts for transesterification of triglycerides to biodiesel [23][24][25][26][27], but also as a solid support for enzyme immobilization [28][29][30]. In this case, the aim of the immobilization is to create a zeolite-enzyme complex that can be used as biocatalysts for the transesterification of triglycerides to biodiesel.…”

Immobilization of lipase from Candida rugosa on Sepabeads®: the effect of lipase oxidation by periodates

“…The use of immobilized lipases is a possible solution to this problem because the enzyme can be recovered from the product and reused [13]. Immobilization of enzyme also enhances its thermal and chemical stability and allows heterogeneous catalysis of enzymatic reactions [14,15]. The concept of immobilizing enzymes on insoluble supports has been the subject of considerable research for over 30 years and consequently, many different methodologies and a vast range of applications have been suggested.…”

Reversible immobilization of Candida rugosa lipase on fibrous polymer grafted and sulfonated p(HEMA/EGDMA) beads

“…Interestingly, immobilization can dramatically affect enzyme properties such as pHdependence, temperature profile, resistance to proteolytic digestion and denaturants, as well as kinetics, and has often been observed to enhance thermostability. A large range of polymer matrices has been employed for lipase immobilization, such as on polysulfone membranes [8,9], acrylate based polymers [10][11][12], and inorganic materials [13]. To optimize the catalytic efficiency and stability of the resulting enzyme-polymer, the attempt was made to understand the immobilization effects on enzymatic properties.…”