Isolation and Characterization of Alkaline Proteases Producing Indigenous Bacillus sp. as a Source of Thrombolytic and Fibrinolytic Agents

T. Akhtar, M. A. A. Sikder, Tania Yeasmin, Md Arafat Al Mamun, M. Hoq, Md Mustafizur Rahman, Md Rashedul Haque, Md Abdul Mazid
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Abstract

Fibrinolytic enzymes derived from bacteria can be effective as a potent, safe and cost-effective thrombolytic agent for the prevention and treatment of cardiovascular diseases. In this study, microorganisms were isolated from various soil samples and screened for their protease activity and protein concentration. Isolates with higher protease activity were subsequently tested for their thrombolytic and fibrinolytic activity. Of a total 37 isolates, 7 exhibited significant zone in SMA media (zone ratio above 1.3). Of these, 8 isolates showed increased protease activity with values ranging from 120 to 199 U/ml. The highest protease activity was observed for the isolate GST12 obtained from mung bean (green gram) soil. The freeze-dried concentrated crude enzymes were screened by in vitro clot lysis method. Six of these eight concentrated enzymes demonstrated clot lysis above 20%, whereas, enzymes from GST12 and GST21 exhibited the highest clot lysis activity (about 27%), which is comparable with standard drug streptokinase. These two enzymes are also strongly associated with fibrinolytic activity. Our findings identify 8 isolates having potential thrombolytic and fibrinolytic activities that belong to Gram-positive Bacillus. Dhaka Univ. J. Pharm. Sci. 22(2): 125-135, 2023 (December)
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可作为血栓溶解剂和纤维蛋白溶解剂来源的产碱性蛋白酶本地芽孢杆菌的分离和特征描述
从细菌中提取的纤溶酶可作为一种有效、安全和经济的溶栓剂,用于预防和治疗心血管疾病。本研究从各种土壤样本中分离出微生物,并对其蛋白酶活性和蛋白质浓度进行筛选。随后,对蛋白酶活性较高的分离物进行了血栓溶解和纤维蛋白溶解活性测试。在总共 37 个分离菌株中,有 7 个在 SMA 培养基中显示出明显的区域(区域比高于 1.3)。其中,8 个分离菌株的蛋白酶活性有所提高,数值从 120 U/ml 到 199 U/ml 不等。从绿豆(青稞)土壤中分离出的 GST12 蛋白酶活性最高。用体外凝块裂解法对冷冻干燥的浓缩粗酶进行了筛选。在这八种浓缩酶中,有六种的血块裂解率超过 20%,而 GST12 和 GST21 的酶表现出最高的血块裂解活性(约 27%),与标准药物链激酶相当。这两种酶也与纤维蛋白溶解活性密切相关。我们的研究结果确定了 8 种具有潜在血栓溶解和纤维蛋白溶解活性的分离物,它们都属于革兰氏阳性芽孢杆菌。Dhaka Univ.22(2):125-135,2023 年(12 月)
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