Immobilized Biocatalysts of Eversa® Transform 2.0 and Lipase from Thermomyces Lanuginosus: Comparison of Some Properties and Performance in Biodiesel Production

Abstract

<jats:p>Eversa® Transform (ET), and the lipase from Thermomyces lanuginosus (TLL), liquid commercial lipases formulations, have been immobilized on octyl agarose beads and their stabilities were compared. Immobilized and free ET forms were more thermostable than TLL formulations at pH 5.0, 7.0, and 9.0, and the ET immobilized form was more stable in the presence of 90% methanol or dioxane at 25 °C and pH 7. Specific activity versus p-nitrophenyl butyrate was higher for ET than for TLL. However, after immobilization the differences almost disappeared because TLL was very hyperactivated (2.5-fold) and ET increased the activity only by 1.6 times. The enzymes were also immobilized in octadecyl methacrylate beads. In both cases, the loading was around 20 mg/g. In this instance, activity was similar for immobilized TLL and ET using triacetin, while the activity of immobilized ET was lower using (S)-methyl mandelate. When the immobilized enzymes were used to produce biodiesel from sunflower oil and methanol in tert-butanol medium, their performance was fairly similar.</jats:p>

Multi-Combilipases: Co-Immobilizing Lipases with Very Different Stabilities Combining Immobilization via Interfacial Activation and Ion Exchange. The Reuse of the Most Stable Co-Immobilized Enzymes after Inactivation of the Least Stable Ones

“…This could be caused by an increase in the diffusional problems of the substrate to reach the enzymes under the PEI layer, as LEU will reduce the mobility of the polymeric bead at least in the area where the enzyme is immobilized increasing the tortuosity of the substrate path towards the immobilized enzyme active center [105][106][107][108][109][110][111]. Moreover, the pore diameter will be also reduced [119]. If the substrate was any of the methyl mandelate esters, the activity of the final biocatalysts was slightly reduced: LEU was not very active versus this substrate and their immobilization will make CALB less accessible to this substrate, the most active enzyme with it.…”

Immobilization of Eversa® Transform via CLEA Technology Converts It in a Suitable Biocatalyst for Biolubricant Production Using Waste Cooking Oil

“…Eversa is a genetically modified industrial evolution of the lipase from Thermomyces lanuginosus (TLL) expressed in Aspergillus oryzae by Novozymes A/S [ 24 , 30 ]. Although both enzymes are similar in terms of primary sequence, a recent comparison between them showed that the functional properties could be fairly different [ 31 ]. Eversa has shown improved thermal stability under different conditions and lower sensibility to the presence of phosphate anions [ 31 ].…”

“…Although both enzymes are similar in terms of primary sequence, a recent comparison between them showed that the functional properties could be fairly different [ 31 ]. Eversa has shown improved thermal stability under different conditions and lower sensibility to the presence of phosphate anions [ 31 ]. However, immobilized Eversa and TLL were similar in the production of biodiesel [ 31 ].…”

“…Eversa has shown improved thermal stability under different conditions and lower sensibility to the presence of phosphate anions [ 31 ]. However, immobilized Eversa and TLL were similar in the production of biodiesel [ 31 ]. In addition, TLL has been successfully applied in the production of synthetic biolubricants.…”

Immobilized Forms of the Ophiostoma piceae Lipase for Green Synthesis of Biodiesel. Comparison with Eversa Transform 2.0 and Cal A

“…In the last years, enzymes like Candida antarctica lipase A (Cal A) [37] and very specially Eversa Transform ® , a genetically-modified variant of the lipase from Thermomyces lanuginosus designed by Novozymes for the synthesis of biodiesel, have been used in this application [36,[38][39][40][41][42]. Both biocatalysts have been immobilized by hydrophobicity on octyl agarose beads [24,39,43], Cal A was also covalently attached to chitosan-coated magnetic nanoparticles activated with GA [44], and mCLEAs of Eversa Transform ® were prepared and tested to produce biodiesel [41]. The recombinant versatile lipase from Ophiostoma piceae (OPEr) has shown to be a robust biocatalyst, whose activity is maintained or improved upon immobilization.…”