Taxonomic group: fungi / Basidiomycota, Ascomycota, Ascomycota
(Phylum: Basidiomycota, Ascomycota, Ascomycota)
The structure was elucidated in this paperNCBI PubMed ID: 30966558Publication DOI: 10.3390/polym10050524Journal NLM ID: 101545357Publisher: Basel: MDPI
Correspondence: Höck H <H.Hoeck
biotec.rwth-aachen.de>; Schwaneberg U <U.Schwaneberg
biotec.rwth-aachen.de>; Engel S <Engel
dwi.rwth-aachen.de>; Keul H <Keul
dwi.rwth-aachen.de>; Möller M <Moeller
dwi.rwth-aachen.de>; Bocola M <M.Bocola
biotec.rwth-aachen.de>; Weingarten S <Simone.weingarten
rwth-aachen.de>
Institutions: DWI—Leibniz Institute for Interactive Materials and Institute of Biotechnology, RWTH Aachen University, Aachen, Germany, DWI—Leibniz Institute for Interactive Materials and Institute of Technical and Macromolecular Chemistry, RWTH Aachen University, Aachen, Germany
Enzyme-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.
immobilization, enzyme engineering, Candida antarctica lipase B, enzymatic ring-opening polymerization, microgel
Structure type: oligomer
Location inside paper: Fig. 1
Aglycon: (->4) Asn-X-Ser/Thr (CalB)
Compound class: N-glycan
Contained glycoepitopes: IEDB_135813,IEDB_137340,IEDB_141807,IEDB_151531,IEDB_153212,SB_74,SB_85
Methods: PCR, genetic methods, MALDI-TOF, flow cytometry, centrifugationmolecular modeling
Comments, role: N-glycosylation site Asn74
3D data: computer modeling
NCBI Taxonomy refs (TaxIDs): 84753,
4932,
4922Reference(s) to other database(s): GTC:G42666HT
Show glycosyltransferases
There is only one chemically distinct structure:
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(later renamed to: Moesziomyces antarcticus)