Strain improvement, optimization and purification studies for enhanced production of streptokinase from Streptococcus uberis TNA-M1
Strain improvement, optimization and purification studies for enhanced production of streptokinase from Streptococcus uberis TNA-M1作者机构：School of Biosciences and Technology VIT University Vellore-632014 TN India
出 版 物：《Frontiers in Biology》 (生物学前沿（英文版）)
年 卷 期：2017年第12卷第5期
页 面：376-384页
核心收录：
学科分类：0710[理学-生物学] 07[理学] 09[农学]
基 金：We are greatly indebted to Vellore Institute of Technology for the constant encouragement help and support for extending necessary facilities
主 题：Streptokinase Streptococcus uberis clot busters mutagenesis optimization
摘 要：BACKGROUND： Screening of isolates for their potency to produce streptokinase was an important criterion of this research. The current study emphasizes the strain improvement, optimization and purification studies for enhanced production of streptokinase from Streptococcus uberis TNA-M1 isolated from bovine milk. METHODS： The study was carried out on samples collected from milk sample. Primary screening and characterization is used as an excellent source for the isolation of 13-hemolytic organisms. Strain improvement was done by both physical ＆ chemical mutagenesis. The enzyme activity was checked by clot lysis assay and confirmed by fibrin plate method. The partially purified and crude enzyme were analysed by high-performance liquid chromatography. Molecular weight ＆ enzyme purity was checked by SDS -PAGE, further confirmed by fibrin zymography. RESULTS： Out of the 3 isolated strains, only one isolate expressed 13-haemolysis with streptokinase （SK） activity. Based on the results of radial caseinolytic assay and blood clot dissolving assay, isolate TNA-M1 demonstrated the highest streptokinase activity. Based on morphological, biochemical and molecular characterization, it was identified as Streptococcus uberis and the strain was named as Streptococcus uberis TNA-M1. The results indicated that ultra-violet （UV） and ethyl methane sulfonate （EMS） were effective mutagenic agents for strain improvement of Streptococcus uberis TNA-M1 and enhanced SK productivity. HPLC analysis was performed in order to confirm the presence of streptokinase with the similar retention time （0.875 min） with its standard （0.854） min. SDS-PAGE of the enzyme showed protein band of approximately 47 kDa and confirmed by fibrin zymography. It exhibited fibrinolytic activity, which was more potent than other fibrinolytic enzymes. Glucose and peptone were recorded to be the optimum carbon and nitrogen sources respectively. CONCLUSION： Thus this study presents its novelty by highlighting the potentia
维普期刊数据库