Yoshimura F, Nishikata M, Suzuki T, Hoover C I, Newbrun E
Arch Oral Biol. 1984;29(7):559-64. doi: 10.1016/0003-9969(84)90078-5.
A trypsin-like, membrane-bound protease from Bacteroides gingivalis was solubilized by Triton X-100 and partially purified by a combination of DEAE-Sepharose and aminophenylmercuric Sepharose chromatography, by taking advantage of the thiol group on the enzyme. The purified enzyme hydrolysed the synthetic substrates benzoyl-L-arginine-p-nitroanilide (L-BAPA), benzoyl-D,L-arginine-beta-naphthylamide (BANA) and tosyl-L-arginine methyl ester, as well as bovine serum albumin and ovalbumin, but not tosyl-L-lysine methyl ester. The enzyme activity was enhanced by SH-reagents and was inhibited to different degrees by SH-inhibitors, chelators and microbial low-molecular-weight inhibitors such as leupeptin, antipain and chymostatin. These microbial inhibitors could be of practical use as ligands for affinity chromatography for further purification. The possible involvement of the protease in periodontal diseases is also discussed.
牙龈卟啉单胞菌中一种类胰蛋白酶的膜结合蛋白酶，利用Triton X - 100使其溶解，并通过DEAE - 琼脂糖和氨基苯汞琼脂糖层析相结合的方法，借助该酶上的巯基进行部分纯化。纯化后的酶可水解合成底物苯甲酰 - L - 精氨酸 - 对硝基苯胺（L - BAPA）、苯甲酰 - D,L - 精氨酸 - β - 萘酰胺（BANA）和甲苯磺酰 - L - 精氨酸甲酯，以及牛血清白蛋白和卵清蛋白，但不能水解甲苯磺酰 - L - 赖氨酸甲酯。该酶的活性可被巯基试剂增强，并受到巯基抑制剂、螯合剂以及诸如亮抑酶肽、抗蛋白酶和抑糜酶素等微生物低分子量抑制剂不同程度的抑制。这些微生物抑制剂可作为亲和层析的配体用于进一步纯化。文中还讨论了该蛋白酶可能与牙周疾病的关系。
