title: In-gel determination of L-amino acid oxidase activity based on the visualization of Prussian blue-forming reaction.
abstract: L-amino acid oxidase (LAAO) is attracting increasing attention due to its important functions. Diverse detection methods with their own properties have been developed for characterization of LAAO. In the present study, a simple, rapid, sensitive, cost-effective and reproducible method for quantitative in-gel determination of LAAO activity based on the visualization of Prussian blue-forming reaction is described. Coupled with SDS-PAGE, this Prussian blue agar assay can be directly used to determine the numbers and approximate molecular weights of LAAO in one step, allowing straightforward application for purification and sequence identification of LAAO from diverse samples.
authors: College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou, China. zhiliangyu76@gmail.com
authors: Zhiliang
authors: Z
authors: Yu
authors: Ning
authors: N
authors: Zhou
authors: Chuntian
authors: C
authors: Zhao
authors: Juanping
authors: J
authors: Qiu
mesh_terms: D002623:Chemistry Techniques, Analytical / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006861:Hydrogen Peroxide / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D050830:L-Amino Acid Oxidase / Q000378:metabolism*; D010084:Oxidation-Reduction; D011540:Prussian Blue Reaction; D013379:Substrate Specificity
chemical_list: D006861:Hydrogen Peroxide; D050830:L-Amino Acid Oxidase
references: J Exp Biol. 2005 Sep;208(Pt 18):3609-22
references: 16155232
references: Nature. 1970 Aug 15;227(5259):680-5
references: 5432063
references: J Bacteriol. 2006 Apr;188(7):2493-501
references: 16547036
references: FEBS Lett. 2002 Nov 20;531(3):509-12
references: 12435602
references: Anal Chem. 1971 Jun;43(7):880-2
references: 5576608
references: PLoS One. 2012;7(3):e33639
references: 22438972
references: Microbiol Immunol. 2007;51(9):889-92
references: 17895606
references: Eur J Biochem. 1993 Jul 15;215(2):351-60
references: 8344302
references: Toxicon. 2001 Dec;39(12):1827-33
references: 11600144
references: Appl Microbiol Biotechnol. 2008 Jul;79(6):925-30
references: 18504575
references: Biofouling. 2011 Mar;27(3):287-93
references: 21390913
references: Can J Microbiol. 2009 Sep;55(9):1096-102
references: 19898552
references: Appl Biochem Biotechnol. 2012 May;167(1):1-13
references: 22367642
references: Appl Environ Microbiol. 2005 Jul;71(7):3551-5
references: 16000761
references: Fish Shellfish Immunol. 2007 Dec;23(6):1178-86
references: 17964810
references: J Microbiol Methods. 2011 Jun;85(3):228-9
references: 21397638
references: Biochemistry. 2006 Dec 26;45(51):15844-52
references: 17176107