Purification and partial characterization of an L-amino acid oxidase from bushmaster snake (Surucucu Pico de Jaca) Lachesis muta muta venom.
1. L-amino acid oxidase (L-AO) from the venom of Lachesis muta muta was purified 72 times (38%) by gel filtration on Sephadex G-100, followed by ion exchange chromatography on DEAE-cellulose and gel filtration on Sephacryl S-300. 2. The protein was shown to be homogeneous by polyacrylamide gel electrophoresis at pH 8.5, immunoelectrophoresis, immunodiffusion and isoelectric focusing. Its specific activity was 44.4 units/mg protein, using 7.5 mM L-leucine as substrate and O-dianisidine as electron donor, at pH 7.6 and 25 degrees C. The increase in absorbance at 436 nm was recorded. 3. The enzyme was characterized as a glycoprotein with an S20,w = 6.72, MW = 138,000 and pI = 5.2. It presented maxima at 389 and 460 nm and contained 2 mol of FAD per mole protein.
Sánchez
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EO
Centro de Pesquisa e Desenvolvimento, Fundação Ezequiel Dias, Belo Horizonte, MG, Brasil.
Magalhães
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A
D000594:Amino Acid Oxidoreductases / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003435:Crotalid Venoms / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D005779:Immunodiffusion; D007122:Immunoelectrophoresis; D007525:Isoelectric Focusing; D050830:L-Amino Acid Oxidase; D012911:Snakes; D013997:Time Factors
D003435:Crotalid Venoms; D000594:Amino Acid Oxidoreductases; D050830:L-Amino Acid Oxidase