Isolation and characterization of extracellular pectin lyase from Penicillium canescens.
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.
Sinitsyna
O A
OA
Chemical Faculty, Lomonosov Moscow State University, Moscow 119992, Russia. oasinitsyna@hotmail.com
Fedorova
E A
EA
Semenova
M V
MV
Gusakov
A V
AV
Sokolova
L M
LM
Bubnova
T M
TM
Okunev
O N
ON
Chulkin
A M
AM
Vavilova
E A
EA
Vinetsky
Y P
YP
Sinitsyn
A P
AP
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
D004271:DNA, Fungal; D005656:Fungal Proteins; D010368:Pectins; D011133:Polysaccharide-Lyases; C027349:pectin lyase; D002118:Calcium