小檗种子蛋白提取物中具有抗菌活性的蛋白酶抑制剂的分离与鉴定

Rutwika S. Thete, S. Roushani, F. Shaikh, Jyoti Kulkarni, R. H. L
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摘要

本研究旨在从茜草(Albizia lebbeck)种子蛋白提取物中鉴定具有抗菌活性的蛋白酶抑制剂(PIs),这些抑制剂可能是克服现有抗菌药物多重耐药、毒性和副作用的天然替代品。从白参种子中提取粗pi,在1% PVP条件下,用硫酸铵(NH4)2SO4分馏提纯部分pi。总蛋白含量在0 ~ 30% (NH4)2SO4饱和蛋白组分F1中较高(7.3±0.17 mg/ml)。琼脂孔扩散法和体外溶液法测定了60 - 90% (NH4)2SO4饱和组分F3对胰蛋白酶具有较高的PI活性。F3蛋白的电泳分析显示,凝胶上有9条具有不同迁移率的条带。不同浓度的F3- 60 - 90% (NH4)2SO4饱和的PIs在琼脂平板上存在抑制区(ZOI),对大肠杆菌、金黄色葡萄球菌和铜绿假单胞菌的抑菌活性分别为100±5µg/ml、100±4µg/ml和90±3µg/ml。我们的研究结果表明,从sirisa种子中提取的pi对所测试的微生物显示出有效的抗菌活性,可以在未来的研究中进一步用于设计或配制天然抗菌药物来治疗微生物感染相关疾病。
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Isolation and Characterization of Protease Inhibitors with Antimicrobial Activity from Sirisa (Albizia lebbeck) Seed Protein Extract
The present study aimed to identify protease inhibitors (PIs) with antimicrobial activity from sirisa (Albizia lebbeck) seed protein extracts that may be a natural alternative to overcome multi-drug resistance, toxicity, and side effects of existing antimicrobial drugs. The crude PIs were extracted from seeds of A. lebbeck in 1% PVP and further partially purified by ammonium sulphate (NH4)2SO4 fractionation. The total protein content was found to be high in 0-30 % (NH4)2SO4 saturated protein fraction F1 (7.3 ± 0.17 mg/ml). Reasonably high PI activity towards trypsin was observed in 60–90 % (NH4)2SO4 saturated fraction F3 assessed by the agar well diffusion method and in vitro solution assay. Electrophoretic profiling of proteins from the F3 fraction showed nine bands on the gel with differential mobility. The presence of a zone of inhibition (ZOI) for different concentrations of F3- 60–90 % (NH4)2SO4 saturated PIs on agar plate demonstrated antimicrobial activity against E.coli, S. aureus, and P. aeruginosa with MIC values of 100 ± 5 µg/ml, 100 ± 4 µg/ml, and 90 ± 3 µg/ml respectively. Our results indicate that PIs from seeds of sirisa display potent antimicrobial activity against the tested microorganisms and could be investigated further in the future use in designing or formulating natural antimicrobial drugs to treat microbial infection-related diseases.
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