Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 4 de 4
Filter
Add more filters










Database
Language
Publication year range
1.
Prikl Biokhim Mikrobiol ; 51(5): 502-10, 2015.
Article in Russian | MEDLINE | ID: mdl-26596087

ABSTRACT

Recombinant strains of Penicillium canescens producing homologous pectin lyase A and heterologous endo- 1,5-α-arabinase A and endo- 1,4-α-polygalacturonase, as well as enzymes of the host strain (α-L-arabinofuranosidases, xylanases, and others), were obtained by genetic engineering. The enzyme preparations (EPs) obtained from the cultural medium of recombinant P. canescens strains efficiently hydrolyzed raw plant material with a high content of pectin compounds. It was shown that the yield of reducing sugars and arabinose increased 16 and 22% in comparison with the control EP based on the host strain when one of the obtained EPs was used for beet pulp hydrolysis. It was established that the most active EP consisted of pectin lyase (10%), endo-1,5-arabinase (26%), α-L-arabinofuranosidase and arabinoxylan-arabinofuranohydrolase (12%), and xylanase (10%). The activities of pectin lyase, polygalacturonase, and arabinase of the EP in reactions with various substrates were determined. The specificity, pH and T-optima, and thermal stability of the homogenous recombinant endo- 1,5-α-arabinase were investigated. The kinetic parameters (K(m), K(cat)) of the linear arabinan hydrolysis were determined.


Subject(s)
Genetic Engineering , Glycoside Hydrolases/biosynthesis , Penicillium/enzymology , Polysaccharide-Lyases/biosynthesis , Glycoside Hydrolases/genetics , Hydrolysis , Pectins/metabolism , Penicillium/genetics , Polysaccharide-Lyases/genetics
2.
Prikl Biokhim Mikrobiol ; 51(4): 402-11, 2015.
Article in Russian | MEDLINE | ID: mdl-26353405

ABSTRACT

Based on the fungus Penicillium verruculosum, we created strains with a complex of extracellular enzymes that contains both cellulolytic enzymes of the fungus and heterologous pectin lyase A from P. canescens and endo- 1,4-α-polygalacturonase from Aspergillus niger. The endopolygalacturonase and pectin lyase activities of enzyme preparations obtained from culture media of the producer strains reached 46-53 U/mg of protein and 1.3-2.3 U/mg of protein, respectively. The optimal temperature and pH values for recombinant pectin lyase and endopolygalacturonase corresponded to those described in the literature for these enzymes. The content of heterologous endopolygalacturonase and pectin lyase in the studied enzyme preparations was 4-5% and 23% of the total protein content, respectively. The yield of reducing sugars upon the hydrolysis of sugar beet and apple processing wastes with the most efficient preparation was 41 and 71 g/L, respectively, which corresponded to a polysaccharide conversion of 49% and 65%. Glucose was the main product of the hydrolysis of sugar beet and apple processing wastes.


Subject(s)
Metabolic Engineering , Penicillium/genetics , Polygalacturonase/genetics , Polysaccharide-Lyases/genetics , Aspergillus niger/enzymology , Aspergillus niger/genetics , Beta vulgaris/chemistry , Glucose/biosynthesis , Glucose/chemistry , Hydrolysis , Malus/chemistry , Pectins/biosynthesis , Pectins/chemistry , Penicillium/enzymology , Polygalacturonase/metabolism , Polysaccharide-Lyases/metabolism , Recombinant Proteins/genetics , Recombinant Proteins/metabolism
3.
Prikl Biokhim Mikrobiol ; 51(6): 584-91, 2015.
Article in Russian | MEDLINE | ID: mdl-26859960

ABSTRACT

The producer of fungal penicillopepsin, an aspartate protease, has been created by genetic engineering. The biochemical and physicochemical properties of the penicillopepsin enzyme preparation obtained from the culture liquid of the producer were studied. Properties of the new enzyme preparation and the commercially available aspergillopepsin were compared. Their proteolytic activities were found to be 670-680 U/g of the preparation. The soluble protein yield upon the wheat flour hydrolysis with penicillopepsin was 2.7 times higher than with aspergillopepsin. It is probably caused by the presence of the xylanase activity in the penicillopepsin preparation.


Subject(s)
Aspartic Acid Endopeptidases/metabolism , Aspartic Acid Proteases/metabolism , Fungal Proteins/metabolism , Penicillium/enzymology , Amino Acid Sequence , Aspartic Acid Endopeptidases/genetics , Aspartic Acid Proteases/genetics , Endo-1,4-beta Xylanases/genetics , Endo-1,4-beta Xylanases/metabolism , Escherichia coli/enzymology , Escherichia coli/genetics , Flour/analysis , Fungal Proteins/genetics , Gene Expression , Genetic Engineering , Hydrolysis , Kinetics , Molecular Sequence Data , Penicillium/genetics , Plasmids/chemistry , Plasmids/metabolism , Proteolysis , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Triticum/metabolism
4.
Prikl Biokhim Mikrobiol ; 48(5): 543-9, 2012.
Article in Russian | MEDLINE | ID: mdl-23101393

ABSTRACT

Complex enzymatic preparations demonstrating activities homologous to pectinlyase A and heterologous to endo-1,4-beta-glucanase from Penicilliumverruculosum and beta-glycosidase from Aspergillusniger have been obtained on the basis of recombinant strains of the fungus Penicilliumcanescens. Two approaches were utilized: development of an enzymatic preparation on the basis of a new strain, which produced all three enzymes, and development of an enzymatic preparation via combined cultivation of three strains, each of which produced one of the enzymes.


Subject(s)
Beta vulgaris/metabolism , Cellulase/metabolism , Medical Waste Disposal/methods , Penicillium/enzymology , Polysaccharide-Lyases/metabolism , Base Sequence , Genetic Engineering , Industrial Microbiology/methods , Molecular Sequence Data , Penicillium/genetics
SELECTION OF CITATIONS
SEARCH DETAIL
...