Abstract Glycolate oxidase (GO) is a protein composed of identical subunits of 40 kDa, but often exhibits different pI values. In previous study, we had proved that extracted GO was actually a mixture GO and phe-rich basic protein (BP), and the BP was ignored easily as it migrated towards the cathode in SDS-electrophoresis. In this paper, the mixture of GO and BP with a specific activity of 121 U/min.mg was extracted. The UV spectrum of the mixture showed an absorption peak at 258 nm which is the characteristic adsorption of phenylalanine, suggesting that BP located on the surface of GO. Similar BP also existed in crude protein extracts from several mode species of organism, including Arabidopsis thaliana, Oryza sativa and Escherichia Coli.