Enzymes that hydrolyze fungal cell wall polysaccharides. The carbonhydrate constitution of mycodextranse, an endo-alpha (1 yields 4)-D-glucanase from Pencillium melinii.
The chemical constitution of the carbohydrate portion of mycodextranase, an exocellular endo-alpha(1 yields 4) D-glucanase of Penicillium melinii, has been investigated. At least 80% of the carbohydrate, consisting exclusively of mannose and glucose, is released from protein by treatment of the enzyme with 0.05 M potassium hydroxide plus 1 M sodium borohydride or 0.5 M sodium hydroxide at 50 degrees. There is concomitant destruction of 60% of the threonine and 15% of the serine of the treated enzyme and an increase in absorption, at 241 nm, of the treated protein's spectrum, indicative of an O-glycosidic beta-hydroxyamino acyl linkage between untreated protein and its associated carbohydrate. Mannose is the monosaccharide involved in this linkage. Smith degradation, methylation, and glycosidase digestions of the carbohydrate indicate that it is present in mycodextranase as side chains of mannose, glucosyl alpha(1 yields 2)-mannose, and mannosyl alpha(1 yields 2)-glucosyl alpha(1 yields 2)-mannose units with each enzyme molecule bearing a calculated average of 25 side chains. Separation of pronase glycopeptides by gel filtration on Sephadex G-25 revealed that 96% of the carbohydrate is present in the highest molecular weight fraction which contains 60% of the threonine of mycodextranase but only 3.5% of the aromatic acids judged by its absorbance at 280 nm. Further fractionation of this glycopeptide component on Sephadex G-75 indicates carbohydrate is restricted to two fractions, one containing 71% by weight of the threonine and serine of mycodextranase and 56% of its carbohydrate. These results suggest carbohydrate chains of mycodextranase are clustered in a few threonin-rich regions along the polypeptide chain rather than being separated from each other by nonglycosylated areas.
D000596:Amino Acids / Q000032:analysis; D002473:Cell Wall / Q000378:metabolism*; D002848:Chromatography, DEAE-Cellulose; D002849:Chromatography, Gas; D002854:Chromatography, Paper; D002855:Chromatography, Thin Layer; D005947:Glucose / Q000032:analysis; D006020:Glycopeptides / Q000032:analysis; D006026:Glycoside Hydrolases / Q000032:analysis*; D007527:Isoenzymes; D008320:Maltose / Q000032:analysis; D008353:Mannitol / Q000032:analysis; D008358:Mannose / Q000032:analysis; D008970:Molecular Weight; D010407:Penicillium / Q000201:enzymology*; D011134:Polysaccharides / Q000378:metabolism; D013402:Sugar Alcohols / Q000032:analysis
D000596:Amino Acids; D006020:Glycopeptides; D007527:Isoenzymes; D011134:Polysaccharides; D013402:Sugar Alcohols; D008353:Mannitol; D008320:Maltose; D006026:Glycoside Hydrolases; C021817:mycodextranase; D005947:Glucose; D008358:Mannose