Publication Details

Category Text Publication
Reference Category Journals
DOI 10.1128/JB.00477-13
Title (Primary) Evidence for benzylsuccinate synthase subtypes obtained by using stable isotope tools
Author Kümmel, S.; Kuntze, K.; Vogt, C.; Boll, M.; Heider, J.; Richnow, H.H.
Source Titel Journal of Bacteriology
Year 2013
Department ISOBIO
Volume 195
Issue 20
Page From 4660
Page To 4667
Language englisch
UFZ wide themes ru3
Abstract

We studied the benzylsuccinate synthase (Bss) reaction mechanism with respect to the hydrogen-carbon bond cleavage at the methyl group of toluene by using different stable isotope tools. Λ values (slopes of linear regression curves for carbon and hydrogen discrimination) for two-dimensional compound-specific stable isotope analysis (2D-CSIA) of toluene activation by Bss-containing cell extracts (in vitro studies) were found to be similar to previously reported data from analogous experiments with whole cells (in vivo studies), proving that Λ values generated by whole cells are caused by Bss catalysis. The Bss enzymes of facultative anaerobic bacteria produced smaller Λ values than those of obligate anaerobes. In addition, a partial exchange of a single deuterium atom in benzylsuccinate with hydrogen was observed in experiments with deuterium-labeled toluene. In this study, the Bss enzymes of the tested facultative anaerobes showed 3- to 8-fold higher exchange probabilities than those for the enzymes of the tested obligate anaerobic bacteria. The phylogeny of the Bss variants, determined by sequence analyses of BssA, the gene product corresponding to the α subunit of Bss, correlated with the observed differences in Λ values and hydrogen exchange probabilities. In conclusion, our results suggest subtle differences in the reaction mechanisms of Bss isoenzymes of facultative and obligate anaerobes and show that the putative isoenzymes can be differentiated by 2D-CSIA.

Persistent UFZ Identifier https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=14168
Kümmel, S., Kuntze, K., Vogt, C., Boll, M., Heider, J., Richnow, H.H. (2013):
Evidence for benzylsuccinate synthase subtypes obtained by using stable isotope tools
J. Bacteriol. 195 (20), 4660 - 4667 10.1128/JB.00477-13