title: Structural analysis of the pehA gene and characterization of its protein product, endopolygalacturonase, of Erwinia carotovora subspecies carotovora.
abstract: A clone producing a polygalacturonase (EC 3.2.1.15) in Escherichia coli was isolated from a genomic library of Erwinia carotovora subspecies carotovora constructed in PUC18. The DNA segment carrying the corresponding structural gene, named pehA, contained an open reading frame (ORF) encoding a 402-amino-acid (aa) polypeptide with an Mr of 42,849. In E. carotovora the polygalacturonase was synthesized with a 26-aa cleavable signal peptide. The mature 376-aa PehA had a calculated Mr of 40,064 and a pl of 10.19. The pH optimum of the enzyme was about 5.5 and the temperature optimum was in the range 35-45 degrees C. Analysis of the reaction products of polygalacturonic acid hydrolysis indicated that the PehA protein is an endopolygalacturonase. No similarity was observed between the aa sequences of PehA and other pectic enzymes of erwinias. However, substantial similarity was detected within the C-terminal portions of PehA and a previously described tomato polygalacturonase, suggesting that the bacterial and eukaryotic polygalacturonases may have a common origin.
authors.affiliation: Department of Molecular Genetics, Uppsala Centre, Swedish University of Agricultural Sciences.
authors.forename: H T
authors.initials: HT
authors.lastname: Saarilahti
authors.forename: P
authors.initials: P
authors.lastname: Heino
authors.forename: R
authors.initials: R
authors.lastname: Pakkanen
authors.forename: N
authors.initials: N
authors.lastname: Kalkkinen
authors.forename: I
authors.initials: I
authors.lastname: Palva
authors.forename: E T
authors.initials: ET
authors.lastname: Palva
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004885:Erwinia / Q000201:enzymology* / Q000235:genetics; D005798:Genes, Bacterial; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010957:Plasmids; D011096:Polygalacturonase / Q000096:biosynthesis / Q000235:genetics*; D013696:Temperature
chemical_list: D011096:Polygalacturonase