MOLECULAR-CLONING AND NUCLEOTIDE-SEQUENCE OF THE PECTATE LYASE GENE FROM PSEUDOMONAS-MARGINALIS N6301

Pectate lyase was purified approximately 29-fold to electrophoretic ho mogeneity from Pseudomonas marginalis N6301. A pectate lyase (PL; EC4. 2.2.2) gene of the strain was cloned and expressed in Escherichia coli . The nucleotides of the PL gene (pel) were sequenced. An open reading frame that encodes a polypeptide (molecular weight: 40,812) composed of 380 amino acids including a 29 amino acid signal peptide was assign ed. The structural gene of pel consisted of 1140 base pairs. The nucle otide sequence of the 5'-flanking region of pel showed a consensus seq uence of the promoter region of the pectin lyase gene (pnl) in P. marg inalis N6301, a Pribnow box, and a ribosome binding site as found in E . coli.