Doi M, Shioi Y, Sasa T
Arch Biochem Biophys. 1986 Nov 1;250(2):358-63. doi: 10.1016/0003-9861(86)90737-x.
Benzoyl-L-arginine p-nitroanilide hydrolase in the etiolated leaves of Zea mays L. has been purified 1,266-fold by a combination of gel filtration, ion exchange, and hydrophobic chromatography with a recovery of 13%. The specific activity of the purified enzyme is 5.7 units/mg protein. The enzyme is an acidic protein with a pI value of 4.6 and optimum pH of 8.2. The molecular weight of the enzyme was estimated to be 59,000. Substrate inhibition was observed at a concentration higher than 30 microM BAPA and the apparent Km for BAPA was 29 microM at pH 8.0. The enzyme activity was inhibited by sulfhydryl reagents, leupeptin, antipain, and N-tosyl-L-lysine chloromethyl ketone. The inhibitor study suggests that the enzyme belongs to the class of the sulfhydryl proteases.
通过凝胶过滤、离子交换和疏水色谱相结合的方法，玉米黄化叶片中的苯甲酰-L-精氨酸对硝基苯胺水解酶已被纯化了1266倍，回收率为13%。纯化酶的比活性为5.7单位/毫克蛋白质。该酶是一种酸性蛋白质，pI值为4.6，最适pH为8.2。酶的分子量估计为59,000。在高于30 microM BAPA的浓度下观察到底物抑制，在pH 8.0时BAPA的表观Km为29 microM。酶活性受到巯基试剂、亮抑酶肽、抑肽酶和N-对甲苯磺酰-L-赖氨酸氯甲基酮的抑制。抑制剂研究表明该酶属于巯基蛋白酶类。
