000097143 001__ 97143
000097143 005__ 20210902121851.0
000097143 0247_ $$2doi$$a10.1039/d0sc04471g
000097143 0248_ $$2sideral$$a121117
000097143 037__ $$aART-2020-121117
000097143 041__ $$aeng
000097143 100__ $$0(orcid)0000-0001-9478-6750$$aVelasco-Lozano, S.
000097143 245__ $$aSelective oxidation of alkyl and aryl glyceryl monoethers catalysed by an engineered and immobilised glycerol dehydrogenase
000097143 260__ $$c2020
000097143 5060_ $$aAccess copy available to the general public$$fUnrestricted
000097143 5203_ $$aEnzymes acting over glyceryl ethers are scarce in living cells, and consequently biocatalytic transformations of these molecules are rare despite their interest for industrial chemistry. In this work, we have engineered and immobilised a glycerol dehydrogenase from Bacillus stearothermophilus (BsGlyDH) to accept a battery of alkyl/aryl glyceryl monoethers and catalyse their enantioselective oxidation to yield the corresponding 3-alkoxy/aryloxy-1-hydroxyacetones. QM/MM computational studies decipher the key role of D123 in the oxidation catalytic mechanism, and reveal that this enzyme is highly enantioselective towards S-isomers (ee > 99%). Through structure-guided site-selective mutagenesis, we find that the mutation L252A sculpts the active site to accommodate a productive configuration of 3-monoalkyl glycerols. This mutation enhances the kcat 163-fold towards 3-ethoxypropan-1, 2-diol, resulting in a specific activity similar to the one found for the wild-type towards glycerol. Furthermore, we immobilised the L252A variant to intensify the process, demonstrating the reusability and increasing the operational stability of the resulting heterogeneous biocatalyst. Finally, we manage to integrate this immobilised enzyme into a one-pot chemoenzymatic process to convert glycidol and ethanol into 3-ethoxy-1-hydroxyacetone and (R)-3-ethoxypropan-1, 2-diol, without affecting the oxidation activity. These results thus expand the uses of engineered glycerol dehydrogenases in applied biocatalysis for the kinetic resolution of glycerol ethers and the manufacturing of substituted hydroxyacetones. This journal is
000097143 536__ $$9info:eu-repo/grantAgreement/ES/DGA-ERDF/E37-20R$$9info:eu-repo/grantAgreement/ES/DGA-FEDER/Construyendo Europa desde Aragón$$9info:eu-repo/grantAgreement/ES/MCIU/RED2018-102403-T$$9info:eu-repo/grantAgreement/ES/MCIU/RTI2018-093431-B-100$$9info:eu-repo/grantAgreement/ES/MCIU/RTI2018-094398-B-I00$$9info:eu-repo/grantAgreement/ES/MEC/FPU14-04338
000097143 540__ $$9info:eu-repo/semantics/openAccess$$aby-nc$$uhttp://creativecommons.org/licenses/by-nc/3.0/es/
000097143 590__ $$a9.825$$b2020
000097143 591__ $$aCHEMISTRY, MULTIDISCIPLINARY$$b22 / 178 = 0.124$$c2020$$dQ1$$eT1
000097143 592__ $$a3.686$$b2020
000097143 593__ $$aChemistry (miscellaneous)$$c2020$$dQ1
000097143 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000097143 700__ $$aRoca, M.
000097143 700__ $$0(orcid)0000-0002-3256-2089$$aLeal-Duaso, A.$$uUniversidad de Zaragoza
000097143 700__ $$0(orcid)0000-0003-1570-4257$$aMayoral, J.A.$$uUniversidad de Zaragoza
000097143 700__ $$0(orcid)0000-0002-2676-8814$$aPires, E.$$uUniversidad de Zaragoza
000097143 700__ $$aMoliner, V.
000097143 700__ $$0(orcid)0000-0003-0031-1880$$aLópez-Gallego, F.
000097143 7102_ $$12013$$2765$$aUniversidad de Zaragoza$$bDpto. Química Orgánica$$cÁrea Química Orgánica
000097143 773__ $$g11, 44 (2020), 12009-12020$$pChem. sci.$$tChemical Science$$x2041-6520
000097143 8564_ $$s586400$$uhttps://zaguan.unizar.es/record/97143/files/texto_completo.pdf$$yVersión publicada
000097143 8564_ $$s33936$$uhttps://zaguan.unizar.es/record/97143/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000097143 909CO $$ooai:zaguan.unizar.es:97143$$particulos$$pdriver
000097143 951__ $$a2021-09-02-10:29:49
000097143 980__ $$aARTICLE