Serratiopeptidase is an alkaline metalloendopeptidase, which acquired wide significance because of its therapeutic applications. The present study was undertaken for purification, characterization, and structural elucidation of serratiopeptidase produced from Streptomyces hydrogenans var. MGS13. The crude enzyme was purified by precipitating with ammonium sulfate, dialysis, and Sephadex gel filtration, resulting in 34% recovery with a 12% purification fold. The purified enzyme S.AMP13 was spotted as a single clear hydrolytic band on casein zymogram and whose molecular weight was found to be 32 kDa by SDS-PAGE. The inhibitor and stability studies revealed that this enzyme is metalloprotease, thermostable, and alkaline in nature. The maximum serratiopeptidase activity was observed at 37 °C and pH 9.0. The partial amino acid sequence of the purified enzyme S.AMP13 by LC-MS/MS analysis shows the closest sequence similarities with previously reported alkaline metalloendopeptidases. The amino acid sequence alignment of S.AMP13 shared a conserved C-terminus region with peptidase-M10 serralysin superfamily at amino acid positions 128–147, i.e., ANLSTRATDTVYGFNSTAGR revealed that this enzyme is a serralysin-like protease. The kinetic studies of the purified enzyme revealed a Km of 1 mg/mL for its substrate casein and Vmax of 319 U/mL/min. The 3D structure of the purified enzyme was modeled by using SWISS-MODEL, and the quality of the structure was authenticated by assessing the Ramachandran plot using PROCHECK server, which suggested that the enzyme was stable with good quality. Inhibitor, stability, electrophoretic, and bioinformatic studies suggested that the purified enzyme obtained from S. hydrogenans var. MGS13 is a serralysin-like protease.

中文翻译：

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从新来源纯化，鉴定和鉴定沙雷菌素样碱性金属蛋白酶的结构

Serratiopeptidase是一种碱性金属内肽酶，由于其治疗用途而具有广泛的意义。进行本研究的目的是纯化，表征和结构解析从氢链霉菌产生的serratiopeptidase。MGS13。通过用硫酸铵沉淀，透析和Sephadex凝胶过滤来纯化粗酶，得到34％的回收率和12％的纯化倍数。纯化的酶S.AMP13在酪蛋白酶谱图上显示为一条清晰的水解带，通过SDS-PAGE发现分子量为32 kDa。抑制剂和稳定性研究表明，该酶本质上是金属蛋白酶，热稳定和碱性的。在37°C和pH 9.0时观察到最大的serratiopeptidase活性。通过LC-MS / MS分析，纯化的酶S.AMP13的部分氨基酸序列显示与先前报道的碱性金属内肽酶最接近的序列相似性。S.AMP13的氨基酸序列比对与肽酶-M10 serralysin超家族共享一个保守的C末端区域，位于128-147位氨基酸，即ANLSTRATDTVYGFNSTAGR表明该酶是一种Serralysin样蛋白酶。纯化酶的动力学研究表明，其底物酪蛋白的Km为1 mg / mL，Vmax为319 U / mL / min。使用SWISS-MODEL对纯化酶的3D结构进行建模，并通过使用PROCHECK服务器评估Ramachandran图来验证结构的质量，这表明该酶稳定且质量良好。抑制剂，稳定性，电泳，和生物信息学研究表明，从S.hydroans变种获得的纯化酶。MGS13是一种溶血素蛋白蛋白酶。