Sie befinden Sich nicht im Netzwerk der Universität Paderborn. Der Zugriff auf elektronische Ressourcen ist gegebenenfalls nur via VPN oder Shibboleth (DFN-AAI) möglich. mehr Informationen...
The identification and characterization of an oxalyl-CoA synthetase from grass pea ( L.)
Ist Teil von
RSC chemical biology, 2022-03, Vol.3 (3), p.32-333
Ort / Verlag
England: RSC
Erscheinungsjahr
2022
Quelle
EZB Electronic Journals Library
Beschreibungen/Notizen
Oxalic acid is a small metabolite found in many plants. It serves as protection from herbivores, a chelator of metal ions, a regulator of calcium levels, and additional tasks. However, it is also a strong di-carboxylic acid that can compromise plant viability by reducing cellular pH. Several metabolic pathways have evolved to control oxalate levels in plants by enzymatic degradation. Among them is the pathway that utilizes oxalyl-CoA synthetase (OCS, EC 6.2.1.8) and ATP to convert oxalate to oxalyl-CoA. Oxalyl-CoA can then be degraded to CO
2
or utilized as a precursor for the synthesis of other compounds. In grass pea (
Lathyrus sativus
L.), a grain legume grown in Asia and Africa for human and animal consumption, the neurotoxic compound β-
N
-oxalyl-
l
-α,β-diaminopropionic acid (β-ODAP) is synthesized from oxalyl-CoA and
l
-α,β-diaminopropionic acid (
l
-DAPA). Here, we report on the identification and characterization of oxalyl CoA-synthetase from grass pea (
Ls
OCS). The gene encoding
Ls
OCS was amplified from grass pea, and then expressed and purified from
E. coli
cells as an untagged, monomeric protein of 56 kDa. Its catalytic efficiency with oxalate,
K
oxalate
M
= 71.5 ± 13.3 μM,
V
max
= 8.2 ± 0.8 μmole min
−1
mg
−1
, was similar to that of OCS homologs from
Arabidopsis thaliana
(
At
AAE3)
and Medicago truncatula
(
Mt
AAE3). The enzyme was crystalized in complex with AMP and is the first OCS whose structure was determined in the thioester-forming conformation. Finally, we propose that substituting
Ls
OCS with an oxalate oxidase or decarboxylase may reduce the levels of β-ODAP in grass pea.
Kinetic and structural analysis of a novel oxalyl-CoA synthetase isolated from grass pea (
Lathyrus sativus
L.).