Characterization of O‐acetylserine (thiol) lyase from Leucaena leucocephala

• DOI: 10.1096/fasebj.27.1_supplement.580.3
• Publication Date: 2021-06-07T12:15:07Z
• Authors: Jannai T. Yafuso; Vishal Singh Negi; Jon‐Paul Bingham; Dulal Borthakur
• Keywords: Mimosine; Leucaena; Lyase

In plants, the final step of cysteine formation involves addition sulfide to O ‐acetylserine (OAS) and release acetate, catlyzed by (thiol) lyase (OASTL). The purpose this study was isolate characterize recombinant OASTL from Leucaena leucocephala (leucaena), a protein‐rich legume used as fodder. contains toxic, nonprotein amino acid, mimosine, which is also produced OASTL. order understand function enzyme, we first studied synthesis. cDNA for cytosolic obtained through interspecies subtractive hybridization Rapid Amplification Ends. sequence codon optimized expressed in Escherichia coli under control T7 promoter. A 37‐kDa protein isolated vitro enzyme activity assays with Na 2 S OAS substrates where detected quantified using High Performance Liquid Chromatography. K m 1.850±0.4146 mM V max 200.6±19.92 μM minute −1 . Central mimosine biosynthetic pathways Characterization will be essential development toxin‐free leucaena its use widespread forage crop. This research funded NSF Award No. CBET 08–27057.