Discovery of Intermediates of lacZ β-Galactosidase Catalyzed Hydrolysis Using dDNP NMR

Christian Kjeldsen, Jan Henrik Ardenkjær-Larsen, Jens Ø. Duus*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

206 Downloads (Pure)

Abstract

Using dissolution dynamic nuclear polarization, the sensitivity of single scan solution state 13C NMR can be improved up to 4 orders of magnitude. In this study, the enzyme lacZ β-galactosidase from Escherichia coli was subjected to hyperpolarized substrate, and previously unknown reaction intermediates were observed, including a 1,1-linked disaccharide. The enzyme is known for making 1,6-transglycosylation, producing products like allolactose, that are also substrates. To analyze the kinetics, a simple kinetic model was developed and used to determine relative transglycosylation and hydrolysis rates of each of the intermediates, and the novel transglycosylation intermediates were determined as better substrates than the 1,6-linked one, explaining their transient nature. These findings suggest that hydrolysis and transglycosylation might be more complex than previously described.
Original languageEnglish
JournalJournal of the American Chemical Society
Volume140
Issue number8
Pages (from-to)3030-3034
ISSN0002-7863
DOIs
Publication statusPublished - 2018

Fingerprint Dive into the research topics of 'Discovery of Intermediates of lacZ β-Galactosidase Catalyzed Hydrolysis Using dDNP NMR'. Together they form a unique fingerprint.

Cite this