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Synthesis of (S)- and (R)-β-Tyrosine by Redesigned Phenylalanine Aminomutase

Peng, Fei 1; Aliyu, Habibu 1; Delavault, André 1; Engel, Ulrike 1; Rudat, Jens 1
1 Institut für Bio- und Lebensmitteltechnik (BLT), Karlsruher Institut für Technologie (KIT)

Abstract:

Phenylalanine aminomutase from Taxus chinensis (TchPAM) is employed in the biosynthesis of the widely used antitumor drug paclitaxel. TchPAM has received substantial attention due to its strict enantioselectivity towards (R)-β-phenylalanine, in contrast to the bacterial enzymes classified as EC 5.4.3.11 which are (S)-selective for this substrate. However, the understanding of the isomerization mechanism of the reorientation and rearrangement reactions in TchPAM might support and promote further research on expanding the scope of the substrate and thus the establishment of large-scale production of potential synthesis for drug development. Upon conservation analysis, computational simulation, and mutagenesis experiments, we report a mutant from TchPAM, which can catalyze the amination reaction of trans-p-hydroxycinnamic acid to (R)- and (S)-β-tyrosine. We propose a mechanism for the function of the highly conserved residues L179, N458, and Q459 in the active site of TchPAM. This work highlights the importance of the hydrophobic residues in the active site, including the residues L104, L108, and I431, for maintaining the strict enantioselectivity of TchPAM, and the importance of these residues for substrate specificity and activation by altering the substrate binding position or varying the location of neighboring residues. ... mehr


Verlagsausgabe §
DOI: 10.5445/IR/1000145519
Veröffentlicht am 28.04.2022
Cover der Publikation
Zugehörige Institution(en) am KIT Institut für Bio- und Lebensmitteltechnik (BLT)
Publikationstyp Zeitschriftenaufsatz
Publikationsjahr 2022
Sprache Englisch
Identifikator ISSN: 2073-4344
KITopen-ID: 1000145519
Erschienen in Catalysts
Verlag MDPI
Band 12
Heft 4
Seiten Art.-Nr.: 397
Bemerkung zur Veröffentlichung Gefördert durch den KIT-Publikationsfonds
Vorab online veröffentlicht am 01.04.2022
Schlagwörter enantioselectivity; β-amino acid; computational enzyme design; functional residues TchPAM; phenylalanine aminomutase
Nachgewiesen in Dimensions
Scopus
Web of Science
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