Article Figures & Data
Figures
- FIG. 1.
SDS-PAGE analysis of CFEs prepared from an IPTG-induced culture of E. coli pET28-ytjE (lanes: 4, total fraction; 5, soluble fraction; 6, insoluble fraction). CFEs prepared from an IPTG-induced culture of E. coli carrying empty vector (lanes: 1, total fraction; 2, soluble fraction; 3, insoluble fraction) were included as a control. Purified His-tagged YtjE protein is also shown (arrow; lane 7). The molecular mass markers are also indicated (kDa).
- FIG. 2.
Alignment of YtjE amino acid sequence from L. lactis IL1403 with Lcd from S. anginosus, PatB from B. subtilis, PatC from L. delbrueckii, and MalY from E. coli. Boxes indicate identical amino acid residues in all the sequences analyzed. The four residues that are invariant in all aminotransferases (▾) (1) are shown. The Lys233 residue is the potential binding site of PLP. Residues typical for aminotransferases (•) and those typical for trans-sulfuration enzymes (○) are also indicated.
- FIG. 3.
Cysteine desulfhydrase activity by in situ staining. YtjE activity was monitored in Tris-glycine gels under nondenaturing conditions, using l-cysteine as the substrate. Lanes: 1, 5 μg purified recombinant protein; 2, 2.5 μg purified recombinant protein; 3, CFE from E. coli recombinant strain expressing YtjE; 4, CFE from E. coli carrying empty vector; 5, CFE from L. lactis IL1403.
- FIG. 4.
Production of MTL, DMDS, and DMTS by coincubating l-methionine with the purified YtjE protein. Values are the means of three independent measurements. The standard errors of the means are also indicated.
Tables
- TABLE 1.
Effects of inhibitors on C-S lyase activity of YtjE
Inhibitor Inhibitor concn Relative activity (%)a Hydroxylamine 1 mM 1.42 (±0.09) 10 mM 0.37 (±0.03) 3-Methyl-2-benzothiazolinone hydrazone 1 mM 23.64 (±0.59) 10 mM 21.71 (±0.49) Iodoacetic acid 1 mM 24.14 (±1.25) 10 mM 2.49 (±0.28) EDTA 1 mM 115.00 (±2.33) 10 mM 124.00 (±0.38) NaCl 0.4% (wt/vol) 78.00 (±1.06) 4% (wt/vol) 22.00 (±0.05) ↵ a 100% activity is the enzyme activity toward l-cystathionine (2 mM) with no inhibitor added, using the thiol determination assay. Values are the means of three determinations, and the standard deviations are shown in parentheses.
