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Please use this identifier to cite or link to this item: http://hdl.handle.net/10277/832

Authors: Melis, Roberta
Internal Tutor: MOLLA, GIANLUCA
Title: Developing of a novel enantioselective biocatalyst acting on L-amino acids of biotechnological interest through a "semi-rational design" approach supported by computational analyses.
Abstract: The synthesis of optically pure enantiomers is a challenge for traditional chemistry. Enzymes often perform better than inorganic catalysts because of their enantioselectivity, effectiveness and ecological sustainability. However, natural enzymes that evolved under the drive of natural selective pressure for their biological function, are not suitable for most industrial applications 1. For this reason, enzymes must be improved by protein engineering. L-amino acid deaminase from Proteus myxofaciens (PmaLAAD) catalyzes the oxidative deamination of L-amino acids (AAs) to the corresponding -keto acids (KA). This biocatalyst can be used for the setup “green processes” to produce pure KA and D-AAs, which are used as raw materials for the synthesis of drugs and agrochemicals. In this project, several biocatalytic processes based on PmaLAAD have been optimized for the bioconversion, deracemization and stereoinversion of (un)natural L-AAs 2. Several natural or synthetic L-AAs have been used as substratet obtaining conversion yields close to 100%. The knowledge of the 3D structure of PmaLAAD 3 was exploited to produce several enzyme variants possessing animproved activity on L-1-naphthylalanine (L-1-Nal) through a semi-rational design approach supported by computational analysis. The most interesting variant (F318A/V412A/V438P-PmaLAAD) was able to convert D,L-1-Nal with a rate 7.5 fold higher than the wild type.
Keywords: Biotransformation, flavoenzyme, biocatalysis, L-amino acid deaminase, protein engineering
Issue Date: 2018
Language: eng
Doctoral course: Biotecnologie, bioscienze e tecnologie chirurgiche
Academic cycle: 31
Publisher: Università degli Studi dell'Insubria
Citation: Melis, R.Developing of a novel enantioselective biocatalyst acting on L-amino acids of biotechnological interest through a "semi-rational design" approach supported by computational analyses. (Doctoral Thesis, Università degli Studi dell'Insubria, 2018).

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PhD_Thesis_MelisRoberta_part1.pdftesto tesi parte 11,94 MBAdobe PDFavailable from: 10/12/2020 View/Open
PhD_Thesis_MelisRoberta_articolo_BiotechnologyAdvances.pdfarticolo7,76 MBAdobe PDFNot available View/Open
PhD_Thesis_MelisRoberta_part2.pdftesto tesi parte 2820,11 kBAdobe PDFavailable from: 10/12/2020 View/Open
PhD_Thesis_MelisRoberta_articolo_AdvancedSynthesisCatalysis.pdfarticolo6,53 MBAdobe PDFNot available View/Open
PhD_Thesis_MelisRoberta_part3.pdftesto tesi parte 32,5 MBAdobe PDFavailable from: 10/12/2020 View/Open
PhD_Thesis_MelisRoberta_articolo_CatalysisScienceTechnology.pdfarticolo7,76 MBAdobe PDFNot available View/Open
PhD_Thesis_MelisRoberta_part4.pdftesto tesi parte 46,28 MBAdobe PDFavailable from: 10/12/2020 View/Open
PhD_Thesis_MelisRoberta_completa.pdftesto completo tesi30,46 MBAdobe PDFNot available View/Open

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