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来源于添加玉米浆培养液培养的螺旋藻中的纤维蛋白溶解酶。

Fibrinolytic enzyme from Arthrospira platensis cultivated in medium culture supplemented with corn steep liquor.

机构信息

Department of Morphology and Animal Physiology, Federal Rural University of Pernambuco-UFRPE, Dom Manoel de Medeiros Ave., Recife, PE 52171-900, Brazil.

Laboratory of Immunopathology Keizo Asami (LIKA), Federal University of Pernambuco- UFPE, Prof. Moraes Ave. Recife, PE 50670-901, Brazil.

出版信息

Int J Biol Macromol. 2020 Dec 1;164:3446-3453. doi: 10.1016/j.ijbiomac.2020.08.217. Epub 2020 Aug 31.

Abstract

Artrhospira (Spirulina) platensis produced fibrinolytic enzyme under mixotrophic conditions using corn steep liquor (CSL). The enzyme was extracted, purified by combination of two chromatographic techniques and biochemically characterized. Maximum fibrinolytic production (268.14 U mg) was obtained using liquid medium culture composed by 0.2% CLS after 10th day of cultivation. Fibrinolytic activity was higher when extracted by homogenization methods and was purified 32.72-fold with specific activity of 7988 U mg. Fibrin zymography showed an active band, indicated acts as a plasmin-like protein with molecular weight of 72 kDa. Fibrinolytic enzyme have optimum pH of 6.0, stable in the range of 6.0 to 10.0 during 24 h and optimum temperature at 40 °C with a stability below 50 °C. Fibrinolytic enzyme is a serine metalloprotease by to be enhanced by Fe and inhibited by PMSF. The enzyme has higher enzymatic activity than most other fibrinolytic enzymes and is stable at temperature and pH human physiological. Overall, the fibrinolytic enzyme from A. platensis has attractive biochemical properties to potential applications in the treatment of thrombosis.

摘要

节旋藻(螺旋藻)在玉米浆(CSL)的混合营养条件下产生纤溶酶。该酶经过两种色谱技术的组合提取、纯化,并进行了生化特性分析。培养第 10 天,在由 0.2% CLS 组成的液体培养基中培养,可获得最大的纤溶酶产量(268.14 U mg)。通过匀浆法提取的纤溶酶活性更高,经过 32.72 倍的纯化后,比活达到 7988 U mg。纤维蛋白凝胶电泳显示出一个活性带,表明其为一种分子量为 72 kDa 的类纤溶酶蛋白。纤溶酶的最适 pH 值为 6.0,在 24 小时内,其在 pH 值 6.0 到 10.0 范围内稳定,最适温度为 40°C,低于 50°C 时稳定性较好。纤溶酶是一种丝氨酸金属蛋白酶,Fe 可增强其活性,PMSF 可抑制其活性。该酶的酶活比大多数其他纤溶酶都高,并且在人体生理温度和 pH 值下稳定。总的来说,节旋藻的纤溶酶具有吸引人的生化特性,有望应用于血栓症的治疗。

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