title: A pectate lyase from a deep subseafloor Georgenia muralis with unusual molecular characteristics.
abstract: A novel pectate lyase of a deep subseafloor bacterium, Georgenia muralis strain JAM 3H7-3 (JCM19733), was purified to homogeneity from a culture broth by an anion exchange chromatography, followed by heat treatment of the enzyme solution at 60 °C for 30 min, and a gel filtration in the presence of SDS. The purified enzyme (Pel-S2) had a molecular mass of ~51 kDa by SDS-PAGE and ~75 kDa by gel filtration. In contrast, without heat treatment, the purified enzyme in SDS sample buffer was found to consist of 23- and 23.5-kDa polypeptides by SDS-PAGE. The enzyme was gradually inactivated by heat treatment with and without SDS in parallel with a shift of polypeptides molecular masses from 23 and 23.5 to 51 kDa on SDS-PAGE. Pel-S2 degraded pectate optimally at pH 10 in a glycine buffer and temperature of 50 °C. The enzyme showed relatively broad substrate specificity toward pectic acid and pectin.
authors.affiliation: Graduate School of Interdisciplinary New Science, Toyo University, 2100 Kujirai, Kawagoe, Saitama, 350-8585, Japan.
authors.forename: Masato
authors.initials: M
authors.lastname: Sasaki
authors.forename: Osamu
authors.initials: O
authors.lastname: Koide
authors.forename: Tohru
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Ron
authors.initials: R
authors.lastname: Usami
authors.forename: Koki
authors.initials: K
authors.lastname: Horikoshi
mesh_terms: D004269:DNA, Bacterial / Q000235:genetics; D005998:Glycine / Q000737:chemistry; D006094:Gram-Positive Bacteria / Q000201:enzymology* / Q000235:genetics; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D010368:Pectins / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D011133:Polysaccharide-Lyases / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity; D014871:Water Microbiology
chemical_list: D004269:DNA, Bacterial; D010455:Peptides; D012336:RNA, Ribosomal, 16S; D010368:Pectins; D011133:Polysaccharide-Lyases; C023911:pectate lyase; D005998:Glycine; C003181:polygalacturonic acid