Purification, characterization and partial amino acid sequences of a xylanase produced by Penicillium chrysogenum.
An extracellular xylanase (1,4-beta-D-xylan xylanohydrolase, EC 3.2.1.8, endo 1,4-beta-xylanase) was found to be the major protein in the culture filtrate of Penicillium chrysogenum when grown on 1% xylan. In contrast to other microorganism no xylanase multiplicity was found in P. chrysogenum under the conditions used. This enzyme was purified to homogeneity by high performance anion-exchange and size-exclusion chromatography. It had an M(r) of 35,000 as estimated by SDS-PAGE and was shown to be active as a monomer. No glycosylation of the protein could be detected neither by a sensitive glycostain nor by enzymatic deglycosylation studies. The enzyme hydrolyzed oat spelt and birchwood xylan randomly, yielding xylose and xylobiose as major end products. It had no cellulase, CMCase, beta-xylosidase or arabinogalactanase activity but acted on p-nitrophenylcellobioside. The pH and temperature optima for its activity were pH 6.0 and 40 degrees C, respectively. Eight peptides obtained after endoproteinase LysC digestion of xylanase have been sequenced, six of them showed considerable amino acid similarity to glucanases and high M(r)/acidic xylanases from different bacteria, yeasts and fungi.
Haas
H
H
Institut für Mikrobiologie (Medizinische Fakultät), Universität Innsbruck, Austria.
Herfurth
E
E
Stöffler
G
G
Redl
B
B
D000595:Amino Acid Sequence; D002523:Edible Grain / Q000378:metabolism; D043364:Endo-1,4-beta Xylanases; D006026:Glycoside Hydrolases / Q000737:chemistry* / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008666:Metalloendopeptidases; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010408:Penicillium chrysogenum / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid; D013696:Temperature
D006026:Glycoside Hydrolases; D043364:Endo-1,4-beta Xylanases; D008666:Metalloendopeptidases; C039830:peptidyl-Lys metalloendopeptidase