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从 和 中回收和部分纯化纤维蛋白溶解蛋白酶及其提取物的细胞毒性和抗氧化活性。

Recovery and partial purification of fibrinolytic protease from and and cytotoxic and antioxidant activity of their extracts.

机构信息

Postgraduate Program in Biotechnology, Federal University of Amazonas, Manaus, Brazil.

Faculty of Pharmaceutical Sciences, Federal University of Amazonas (UFAM), Manaus, Brazil.

出版信息

Prep Biochem Biotechnol. 2024 Apr;54(4):545-552. doi: 10.1080/10826068.2023.2253466. Epub 2023 Sep 5.

DOI:10.1080/10826068.2023.2253466
PMID:37667995
Abstract

Mushrooms are a source of primary and secondary metabolites. Little is known about the most suitable conditions for production of mushrooms by submerged fermentation. This article reports antioxidant and cytotoxic assays, in addition to quantitatively evaluating the content of proteases with fibrinolytic action in the crude extracts of two species of edible mushrooms produced in different formulations, as well as evaluating the recovery of these enzymes by aqueous two-phase systems (ATPS). The mushrooms and , at concentration of 100 µg/mL, displayed inhibition of DPPH and ABTS radicals below 50%. In the cytotoxicity test, the cells human fibroblast cell lines (MRC-5) showed cell viability greater than 80%. Concerning fibrinolytic activity, presented 226.47 ± 7.26 U/mL, therefore being more efficient than (71.5 ± 0.56 U/mL). In the recovery of the extract by ATPS, the fibrinolytic protease was partitioned in the salt phase (30.25 U/mL). The molecular mass of the proteases was between 75 and 100 kDa. These results prove the low cytotoxicity of the extracts produced and that fermentation in supplemented malt broth favored the excretion of fibrinolytic proteases compared to the other evaluated media.

摘要

蘑菇是初级和次级代谢物的来源。关于液体深层发酵生产蘑菇的最适宜条件知之甚少。本文报告抗氧化和细胞毒性测定,以及定量评估两种不同配方生产的两种可食用蘑菇粗提物中具有纤维蛋白溶解作用的蛋白酶含量,同时评估这些酶通过双水相系统(ATPS)的回收情况。蘑菇在 100μg/mL 浓度下,对 DPPH 和 ABTS 自由基的抑制率低于 50%。在细胞毒性试验中,人成纤维细胞系(MRC-5)的细胞存活率大于 80%。关于纤维蛋白溶解活性,显示出 226.47±7.26 U/mL,因此比(71.5±0.56 U/mL)更有效。在通过 ATPS 回收 提取物时,纤维蛋白溶解蛋白酶分配在盐相(30.25 U/mL)中。蛋白酶的分子量在 75 到 100 kDa 之间。这些结果证明了所产生的提取物的低细胞毒性,并且与其他评估的培养基相比,在补充麦芽肉汤中的发酵有利于纤维蛋白溶解蛋白酶的分泌。

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