Comparison of Candida antarctica Lipase B Variants for Conversion of ε-Caprolactone in Aqueous Medium—Part 2
AbstractEnzyme-catalyzed ring-opening polymerization of lactones is a method of increasing interest for the synthesis of polyesters. In the present work, we investigated which changes in the structure of Candida antarctica lipase B (CaLB) shift the catalytic equilibrium between esterification and hydrolysis towards polymerization. Therefore, we present two concepts: (i) removing the glycosylation of CaLB to increase the surface hydrophobicity; and (ii) introducing a hydrophobic lid adapted from Pseudomonas cepacia lipase (PsCL) to enhance the interaction of a growing polymer chain to the elongated lid helix. The deglycosylated CaLB (CaLB-degl) was successfully generated by site-saturation mutagenesis of asparagine 74. Furthermore, computational modeling showed that the introduction of a lid helix at position Ala148 was structurally feasible and the geometry of the active site remained intact. Via overlap extension PCR the lid was successfully inserted, and the variant was produced in large scale in Pichia pastoris with glycosylation (CaLB-lid) and without (CaLB-degl-lid). While the lid variants show a minor positive effect on the polymerization activity, CaLB-degl showed a clearly reduced hydrolytic and enhanced polymerization activity. Immobilization in a hydrophobic polyglycidol-based microgel intensified this effect such that a higher polymerization activity was achieved, compared to the “gold standard” Novozym® 435. View Full-Text
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Höck, H.; Engel, S.; Weingarten, S.; Keul, H.; Schwaneberg, U.; Möller, M.; Bocola, M. Comparison of Candida antarctica Lipase B Variants for Conversion of ε-Caprolactone in Aqueous Medium—Part 2. Polymers 2018, 10, 524.
Höck H, Engel S, Weingarten S, Keul H, Schwaneberg U, Möller M, Bocola M. Comparison of Candida antarctica Lipase B Variants for Conversion of ε-Caprolactone in Aqueous Medium—Part 2. Polymers. 2018; 10(5):524.Chicago/Turabian Style
Höck, Heidi; Engel, Stefan; Weingarten, Simone; Keul, Helmut; Schwaneberg, Ulrich; Möller, Martin; Bocola, Marco. 2018. "Comparison of Candida antarctica Lipase B Variants for Conversion of ε-Caprolactone in Aqueous Medium—Part 2." Polymers 10, no. 5: 524.
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