Molecular cloning, expression and characterization of a serine proteinase from Japanese edible mushroom, Grifola frondosa: solving the structure - function anomaly of a reported aminopeptidase
نویسنده
چکیده
Abbreviations: CAPS: N-cyclohexyl-3-aminopropanesulfonic acid cDNA: omplementary DNA to mRNA CHES: N-cyclohexyl-2-aminoethanesulfonic acid. DTT: dithiothreitol FPLC: fast protein liquid chromatography IPTG: isopropylthio-β-D-galactoside LB: Luria-Bertani MES: 2-(N-morpholino)ethanesulfonic acid Ni-NTA: Nickel-nitriloacetic acid ORF: open reading frame PCR: polymerase chain reaction PMSF: phenylmethanesulphonylfluoride PVDF: polyvinylidene fluoride RACE: rapid amplification of cDNA ends SDS-PAGE: sodium dodecyl sulfate polyacrylamide gel electrophoresis TAPS: N-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid UTR: untranslated region
منابع مشابه
Characterization, occurrence, and molecular cloning of a lectin from Grifola frondosa: jacalin-related lectin of fungal origin.
A lectin named GFL was isolated from the fruiting body of the basidiomycete mushroom Grifola frondosa, which belongs to Aphyllophorales. The lectin had a molecular mass of 24 kDa on SDS-PAGE. The hemagglutinating activity of GFL was not inhibited by any monosaccharide, and inhibited only by porcine stomach mucin so far as tested. The occurrence of GFL was studied at three stages during fruiting...
متن کاملPurification, Characterization and Antioxidant Activities of Enzymolysis Polysaccharide from Grifola frondosa
Our previous study revealed that the antioxidant activity of polysaccharide (coded as FGFP) extracted from Grifola frondosa by enzymolysis treatment was significantly superior than that (coded as GFP) extracted by boiling-water. In this study, one purified polysaccharide fractions (coded as FGFP-11) was obtained from FGFP by purified using DEAE-52 column and Sephacryl S-500HR column. Results in...
متن کاملAntioxidant properties and antioxidant compounds of various extracts from the edible basidiomycete Grifola frondosa (Maitake).
Grifola frondosa is an edible mushroom currently available in Taiwan. Ethanolic, cold-water and hot-water extracts were prepared and their antioxidant properties were investigated. At 1 mg/mL, G. frondosa T1 and T2 cold-water extracts showed high reducing powers of 1.02 and 0.50, respectively. Chelating abilities on ferrous ions of G. frondosa T1 and T2 were higher for cold-water extracts than ...
متن کاملPurification, Characterization and Antioxidant Activities of Enzymolysis Polysaccharide from Grifola frondosa
Our previous study revealed that the antioxidant activity of polysaccharide (coded as FGFP) extracted from Grifola frondosa by enzymolysis treatment was significantly superior than that (coded as GFP) extracted by boiling-water. In this study, one purified polysaccharide fractions (coded as FGFP-11) was obtained from FGFP by purified using DEAE-52 column and Sephacryl S-500HR column. Results in...
متن کاملCloning and Enhanced Expression of an Extracellular Alkaline Protease from a Soil Isolate of Bacillus clausii in Bacillus subtilis
in the detergent industry. In this study, the extracellular alkaline serine protease gene, aprE, from Bacillusclausii was amplified by PCR and further cloned and expressed in B. subtilis WB600 using the pWB980 expression vector. Protease activity of the recombinant B. subtilis WB600 harboring the plasmid pWB980/aprEreached up to 1020 U/ml, approximately 3-folds higher than the nativ...
متن کامل