Isolation of Homogeneous Polysaccharide Monooxygenases from Fungal Sources and Investigation of Their Synergism with Cellulases when Acting on Cellulose.
Lytic polysaccharide monooxygenases (PMO) discovered several years ago are enzymes classified as oxidoreductases. In nature, they participate in microbial degradation of cellulose together with cellulases that belong to the hydrolytic type of enzymes (class of hydrolases). Three PMO from ascomycetes - Thielavia terrestris, Trichoderma reesei, and Myceliophthora thermophila - were isolated and purified to homogeneous state using various types of chromatography. The first two enzymes are recombinant proteins heterologously expressed by the Penicillium verruculosum fungus, while the third is a native PMO secreted by M. thermophila. When acting on microcrystalline cellulose, all these PMOs displayed synergism with the cellulase complex of the P. verruculosum fungus. Replacing 10% of cellulases (by protein concentration) with PMO in the presence of 6.25 mM gallic acid or 2.5 µM of cellobiose dehydrogenase from M. thermophila, used as electron donors for PMO, resulted in the 17-31% increase in the yield of reducing sugars after 24-48 h of the enzymatic reaction.
Federal Research Center Fundamentals of Biotechnology, Russian Academy of Sciences, Moscow, 119071, Russia.
A G
AG
Bulakhov
A V
AV
Gusakov
A V
AV
Chekushina
A D
AD
Satrutdinov
A V
AV
Koshelev
V Yu
VY
Matys
A P
AP
Sinitsyn
D001203:Ascomycota / Q000201:enzymology; D002237:Carbohydrate Dehydrogenases / Q000378:metabolism; D044602:Cellulases / Q000302:isolation & purification / Q000378:metabolism*; D002482:Cellulose / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D005707:Gallic Acid / Q000737:chemistry; D007700:Kinetics; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010407:Penicillium / Q000201:enzymology / Q000378:metabolism; D010455:Peptides / Q000032:analysis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D005656:Fungal Proteins; D010455:Peptides; D011994:Recombinant Proteins; D005707:Gallic Acid; D002482:Cellulose; D006899:Mixed Function Oxygenases; D002237:Carbohydrate Dehydrogenases; C019859:cellobiose-quinone oxidoreductase; D044602:Cellulases; C109691:microcrystalline cellulose