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        Droplet microfluidics for double lipase immobilisation using TiO2 and alginate microbeads

        Rosilene Andrea Welter,Harrson Silva Santana,Bruna Gregatti Carvalho,Natalia Melani,Michael Oelgemöller,Lucimara Gaziola de la Torre,Osvaldir Pereira Taranto 한국공업화학회 2022 Journal of Industrial and Engineering Chemistry Vol.110 No.-

        A double immobilisation technique was developed for C. antarctica lipase (CALB) that improved its enzymaticactivity and solved difficulties frequently observed in reactions catalysed by lipases. The firstimmobilisation consisted of CALB adsorption onto a TiO2 nanoparticle surface (CALB TiO2). The adsorptionwas carried out by an oriented monolayer formed by CALB’s hydrophilic amino acids and the TiO2 surface,leaving the CALB’s active site accessible for reaction. As a result, an increase in enzymatic activity wasachieved. The Relative Enzymatic Activity ((REA) obtained was 289%. The second immobilisation consistedof CALBTiO2 entrapment into calcium alginate microbeads [(CALBTiO2)EDTA-Ca], obtained by an internalcrosslinking ion-exchange mechanism and using microfluidic droplet technique. The microbiocatalystobtained, (CALBTiO2)EDTA-Ca, retained a high enzymatic activity (REAt=0 = 232%) and stability(REAt=30days = 263%) at a size (diameter, Ø=8.9104 nm) that enabled easier recovery than CALBTiO2(Ø = 2.3102 nm) or CALBFree (Ø=5.0 nm), and showed a favourable porosity for diffusion without releasingCALBTiO2. Although the microbeads showed CALBFree leaching, as demonstrated by the loss of REA after amechanical resistant test, (CALBTiO2)EDTA-Ca maintained an almost constant REA.

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