title: Isolation and characterization of extracellular pectin lyase from Penicillium canescens.
abstract: Pectin lyase A (molecular weight 38 kD by SDS-PAGE, pI 6.7) was purified to homogeneity from culture broth of the mycelial fungus Penicillium canescens using chromatographic techniques. During genomic library screening, the gene encoding pectin lyase A from P. canescens (pelA) was isolated and sequenced, and the amino acid sequence was generated by applying the multiple alignment procedure (360 residues). A theoretical model for the three dimensional structure of the protein molecule was also proposed. Different properties of pectin lyase A were investigated: substrate specificity, pH- and temperature optimum of activity, stability under different pH and temperature conditions, and the effect of Ca2+ on enzyme activity. In the course of the laboratory trials, it was demonstrated that pectin lyase A from P. canescens could be successfully applied to production and clarification of juice.
authors.affiliation: Chemical Faculty, Lomonosov Moscow State University, Moscow 119992, Russia. oasinitsyna@hotmail.com
authors.forename: O A
authors.initials: OA
authors.lastname: Sinitsyna
authors.forename: E A
authors.initials: EA
authors.lastname: Fedorova
authors.forename: M V
authors.initials: MV
authors.lastname: Semenova
authors.forename: A V
authors.initials: AV
authors.lastname: Gusakov
authors.forename: L M
authors.initials: LM
authors.lastname: Sokolova
authors.forename: T M
authors.initials: TM
authors.lastname: Bubnova
authors.forename: O N
authors.initials: ON
authors.lastname: Okunev
authors.forename: A M
authors.initials: AM
authors.lastname: Chulkin
authors.forename: E A
authors.initials: EA
authors.lastname: Vavilova
authors.forename: Y P
authors.initials: YP
authors.lastname: Vinetsky
authors.forename: A P
authors.initials: AP
authors.lastname: Sinitsyn
mesh_terms: D000595:Amino Acid Sequence; D001628:Beverages; D002118:Calcium / Q000494:pharmacology; D002852:Chromatography, Ion Exchange; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005110:Extracellular Space / Q000201:enzymology*; D005638:Fruit / Q000201:enzymology / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis / Q000187:drug effects; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D010368:Pectins / Q000378:metabolism; D010407:Penicillium / Q000201:enzymology* / Q000235:genetics; D011133:Polysaccharide-Lyases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D011487:Protein Conformation; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D004271:DNA, Fungal; D005656:Fungal Proteins; D010368:Pectins; D011133:Polysaccharide-Lyases; C027349:pectin lyase; D002118:Calcium