Žun Gašper, Kos Janko, Sabotič Jerica
Department of Biotechnology, Jožef Stefan Institute, Jamova cesta 39, Ljubljana, Slovenia.
Faculty of Pharmacy, University of Ljubljana, Aškerčeva cesta 7, Ljubljana, Slovenia.
3 Biotech. 2017 Jul;7(3):230. doi: 10.1007/s13205-017-0813-7. Epub 2017 Jul 6.
L-Amino acid oxidases (LAO) are widely distributed enzymes but those from snake venoms have been studied the most. We describe a method for in-gel detection of LAO activities based on HO detection by a horseradish peroxidase-coupled reaction using o-phenylenediamine. Complex substrates and single L-amino acids were used successfully for screening LAO activities in higher fungi using crude aqueous extracts of fruiting bodies of 22 basidiomycetes and 1 ascomycete. Half of these samples exhibited one to two bands of LAO activities with mostly broad substrate specificities and a variety of apparent molecular masses ranging from 25 to 200 kDa that were generally more active at pH 5.5 than at pH 8.0. Mushrooms are shown to be a rich source of LAOs that could find use in various medical and biotechnological applications.
L-氨基酸氧化酶(LAO)是广泛分布的酶,但对来自蛇毒的L-氨基酸氧化酶研究得最多。我们描述了一种基于使用邻苯二胺的辣根过氧化物酶偶联反应检测过氧化氢(HO)来在凝胶中检测LAO活性的方法。复杂底物和单一L-氨基酸已成功用于使用22种担子菌和1种子囊菌子实体的粗水提取物筛选高等真菌中的LAO活性。这些样品中有一半显示出一到两条LAO活性条带,大多具有广泛的底物特异性和各种表观分子量,范围从25至200 kDa,通常在pH 5.5时比在pH 8.0时更具活性。结果表明,蘑菇是LAO的丰富来源,可用于各种医学和生物技术应用。