Purification and biochemical characterization of polygalacturonases produced by Aureobasidium pullulans.
The extracellular polygalacturonases produced by Aureobasidium pullulans isolated from waters of the Danube river were partially purified and characterized. The pH optima of polygalacturonases produced in the first phases of cultivation (48 h) and after 10 d as well as their optima of temperature, thermal stabilities, molecular masses, isoelectric points, action pattern and ability to cleave polymeric and oligomeric substrates were compared. Polygalacturonases with a random action pattern (random cleavage of pectate forming a mixture of galactosiduronides with a lower degree of polymerization) [EC 3.2.1.15] were produced only in the first phases of growth, while exopolygalacturonases [EC 3.2.1.67] with a terminal action pattern (cleavage of pectate from the nonreducing end forming D-galactopyranuronic acid as a product) were found during the whole growth. The main enzyme form with a random action pattern was glycosylated and its active site had the arrangement described previously for the active site of polygalacturonase of phytopathogenic fungi.
Stratilová
Eva
E
Institute of Chemistry, Slovak Academy of Sciences, Dúbravská cesta 9, SK-84238 Bratislava, Slovakia. chemevi@savba.sk
Dzúrová
Mária
M
Breierová
Emília
E
Omelková
Jirina
J
D001203:Ascomycota / Q000201:enzymology* / Q000302:isolation & purification; D018153:Czech Republic; D004795:Enzyme Stability; D006026:Glycoside Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D011096:Polygalacturonase / Q000302:isolation & purification / Q000378:metabolism*; D011489:Protein Denaturation; D013816:Thermodynamics; D014871:Water Microbiology
D006026:Glycoside Hydrolases; D011096:Polygalacturonase; C020136:exopolygalacturonase