Thermostable S-alkylcysteine alpha, beta-lyase from a thermophile: purification and properties

Agric Biol Chem. 1990 Aug;54(8):2069-76.

Abstract

S-Alkylcysteine alpha, beta-lyase was found in a thermophile, Bacillus sp. 41A, which was newly isolated from soil, and purified to homogeneity from the cell extract. The enzyme has a molecular weight of about 76,000, and is composed of two subunits identical in molecular weight (39,000). The enzyme requires pyridoxal 5'-phosphate as a coenzyme, and catalyzes alpha, beta-elimination of S-methyl-L-cysteine and its analogs such as S-ethyl-L-cysteine, L-djenkolate, L-cystine, Se-methyl-L-selenocysteine, and O-methyl-DL-serine. However, S-methyl-D-cysteine, D-cystine, L-methionine, and L-norleucine were inert. The enzyme also catalyzes the beta-replacement reaction of S-methyl-L-cysteine with various thiols to yield the corresponding S-substituted cysteines. In addition to S-methyl-L-cysteine, Se-methyl-L-selenocysteine and O-methyl-DL-serine also serve as beta-substituent acceptors in the beta-replacement reaction. The enzyme is most active at 70 degrees C and stable at high temperatures. Automated Edman degradation provided the N-terminal sequence of the first 44 amino acids. The amino acid sequence in the vicinity of the lysyl residue to which pyridoxal 5'-phosphate is bound, was -Lys-His-Gln-Arg- by Edman degradation of the pyridoxyl peptide obtained by digestion with trypsin after reduction with sodium borohydride.

MeSH terms

  • Amino Acid Sequence
  • Bacillus / enzymology*
  • Binding Sites
  • Carbon-Sulfur Lyases*
  • Culture Media
  • Cysteine / analogs & derivatives
  • Cysteine / pharmacology
  • Enzyme Stability
  • Hot Temperature
  • Lyases / antagonists & inhibitors
  • Lyases / isolation & purification*
  • Lyases / metabolism
  • Molecular Sequence Data
  • Molecular Weight
  • Substrate Specificity

Substances

  • Culture Media
  • S-methylcysteine
  • Lyases
  • Carbon-Sulfur Lyases
  • S-alkylcysteine lyase
  • Cysteine