Isolation and characterization of a serine protease from the sprouts of Pleioblastus hindsii Nakai

• Published in: Phytochemistry (Oxford) 2000-07, Vol.54 (6), p.559-565
• Main Authors: Arima, Kazunari, Uchikoba, Tetsuya, Yonezawa, Hiroo, Shimada, Masayuki, Kaneda, Makoto
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Bamboo sprout; Biological and medical sciences; Chromatography, Agarose; Cucumisin; Electrophoresis, Polyacrylamide Gel; Enzymes; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Gramineae; Hydrogen-Ion Concentration; Hydrolases; Immunoblotting; Metabolism; Molecular Sequence Data; Plant physiology and development; Plant protease; Pleioblastus hindsii; Poaceae - chemistry; Poaceae - enzymology; Serine Endopeptidases - chemistry; Serine Endopeptidases - isolation & purification; Serine protease; Substrate Specificity
• ISSN: 0031-9422; 1873-3700
• DOI: 10.1016/S0031-9422(00)00075-3

An endopeptidase has been purified from sprouts of bamboo ( Pleioblastus hindsii Nakai) to electrophoretic homogeneity by four purification steps. Its M r was estimated to be 82 kDa by SDS–PAGE. Enzyme activity was inhibited strongly by diisopropyl fluorophosphate, and weakly by p-chloromercuriphenylsulfonic acid, but not at all by EDTA or pepstatin, indicating that it was a serine protease. The preferential cleavage sites for this protease were found to be large hydrophobic and amide residues at the P 1 position. The specificity of the bamboo serine protease differed from that of cucumisin [EC 3.4.21.25], which cleaved the charged amino acid residues at the P 1 position.