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具有强大抗癌潜力的L-蛋氨酸酶的分离与鉴定。

Isolation and characterization of L-methioninase with promising a powerful anticancer.

作者信息

Ashkan Mada F, Younis Sadia A, Elazab Nahla T

机构信息

Biological Sciences Department, College of Science & Arts, King Abdulaziz University, Rabigh 21911, Saudi Arabia.

Department of Botany, Molecular Microbial Lab, Faculty of Science, Mansoura University, Egypt.

出版信息

Saudi J Biol Sci. 2023 Dec;30(12):103870. doi: 10.1016/j.sjbs.2023.103870. Epub 2023 Nov 4.

Abstract

Bioactive components derived from medicinal herbs have recently acquired popularity due to their efficacy in treating various ailments, including cancer and infectious diseases. In this study, the anticancer enzyme, L-methioninase isolated from medicinal plants endophytic fungi, evaluated its promising therapeutic agents against different types of human cancers. L methionine was purified using column chromatography with the stationary phase of Sephadex G-200 with 6.6-fold purification, which increased the specific activity of 71.3 U/mg of protein with a recovery rate of 48.2 %. On the SDS-PAGE chromatogram, the apparent molecular mass of the isolated enzyme was 48 kDa, and its highest activity was observed at pH 8 and 35 °C. The enzyme was catalytically stable within the pH range of 6.0-9.0 and below 40 °C. This study demonstrates that isolated L-methioninase is particularly efficient against tumour cell lines in vitro. The crude and purified L-methioninase inhibited 60 and 80 % of the growth of the breast cancer cell line (MCF-7), respectively, with an estimated IC = 12.6 μg/ml (crude) and IC = 5.0 μg/ml for purified L-methioninase from isolate 8 with accession no MZ675362. Because of this, pure L-methioninase has better catalytic characteristics and significant thermal stability, which could be used as a cancer-fighting substance than the enzyme purified from other sources.

摘要

草药衍生的生物活性成分最近因其在治疗各种疾病(包括癌症和传染病)方面的功效而受到欢迎。在本研究中,从药用植物内生真菌中分离出抗癌酶L-蛋氨酸酶,并评估了其作为针对不同类型人类癌症的有前景治疗剂的潜力。使用Sephadex G-200固定相通过柱色谱法纯化L-蛋氨酸,纯化倍数为6.6倍,使比活性提高到71.3 U/mg蛋白质,回收率为48.2%。在SDS-PAGE色谱图上,分离出的酶的表观分子量为48 kDa,在pH 8和35°C时观察到其最高活性。该酶在pH 6.0-9.0范围内和40°C以下具有催化稳定性。本研究表明,分离出的L-蛋氨酸酶在体外对肿瘤细胞系特别有效。粗制和纯化的L-蛋氨酸酶分别抑制乳腺癌细胞系(MCF-7)生长的60%和80%,对于登录号为MZ675362的分离株8的纯化L-蛋氨酸酶,估计IC50 = 12.6 μg/ml(粗制)和IC50 = 5.0 μg/ml。因此,纯L-蛋氨酸酶具有更好的催化特性和显著的热稳定性,与从其他来源纯化的酶相比,它可以用作抗癌物质。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/955d/10663931/85bed54da6e3/gr1.jpg

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