Production of Nattokinase from Bacillus amyloliquefaciens MRS18: A Bacterial Strain Isolated from Fermented Beans.

Simran B Coutinho, Rutuja R Shirodkar, Anju Kanjirakkandy, Vijayaganapathi Arulmozhi, Mohanasrinivasan Vaithilingam, Subathra D Chandrasekaran
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Abstract

Background: Nattokinase (NK) is a naturally occurring fibrinolytic protease enzyme obtained from the traditional Japanese food called Natto and has several uses in the pharmaceutical and medical industries. Nowadays, the most often used thrombolytic agent in the clinical field is NK, in part because it is less expensive than other thrombolytic medicines.

Objectives: The objective of this study is to investigate the screening, isolation and characterization of the NK enzyme-producing Bacillus strain from fermented Soya beans.

Methods: The sample of fermented soya beans were tested for the presence of fibrinolytic protease- producing bacteria, followed by the screening, extraction, characterization and clot lysis assays.

Results: A total of three isolates were screened for caseinolytic activities by casein hydrolysis assay. Out of those isolates, MRS18 was found to be potent in producing the enzyme proteinase. To determine the taxonomy and phylogeny of these isolates, biochemical and molecular characterization has been carried out. Bacillus amyloliquefaciens MRS18 has been found with the highest caseinolytic activity. The clot lysing ability of the potent strain Bacillus amyloliquefaciens was found to be 61.7% after 120 min, and on further purification, by ammonium sulphate precipitation method, the lysis percentage was found to be 656% after 120 min.

Conclusion: From the results of the present study, we concluded that Bacillus amyloliquefaciens isolated from the fermented soya beans produced an NK enzyme that exhibits immense potential to lyse blood clots.

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从发酵豆中分离出的芽孢杆菌 MRS18 生产纳豆激酶
背景:纳豆激酶(NK)是从日本传统食品纳豆中提取的一种天然纤溶蛋白酶,在制药和医疗行业有多种用途。目前,临床上最常用的溶栓药物是 NK,部分原因是它比其他溶栓药物便宜:本研究的目的是从发酵大豆中筛选、分离和鉴定产生 NK 酶的芽孢杆菌菌株:方法:检测发酵大豆样品中是否存在产纤维蛋白溶解酶的细菌,然后进行筛选、提取、定性和凝块裂解试验:结果:通过酪蛋白水解试验,共筛选出三种分离菌具有酪蛋白溶解活性。在这些分离菌中,发现 MRS18 能有效产生蛋白酶。为了确定这些分离物的分类和系统发育,对它们进行了生化和分子鉴定。发现淀粉样芽孢杆菌 MRS18 具有最高的酪蛋白溶解活性。通过硫酸铵沉淀法进一步纯化后,发现 120 分钟后有效菌株淀粉样芽孢杆菌的凝块裂解能力为 61.7%,裂解率为 65.6%:从本研究的结果中,我们得出结论:从发酵大豆中分离出的淀粉样芽孢杆菌产生了一种 NK 酶,这种酶具有溶解血凝块的巨大潜力。
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