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首页> 外文期刊>International Journal of Biological Macromolecules: Structure, Function and Interactions >Purification and characterization of L-glutaminase enzyme from camel liver: Enzymatic anticancer property
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Purification and characterization of L-glutaminase enzyme from camel liver: Enzymatic anticancer property

机译：骆驼肝脏L-谷氨酰胺酶的纯化与表征：酶促抗癌性能

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摘要

L-Glutaminase has gained an important attention as glutamine-depleting enzyme in treatment of various cancers. Therefore, this study aimed to purify, characterize and investigate antitumor activity oft-glutaminase from camel liver mitochondria (CL-Glu), since no available information about CL-Glu from camel. CL-Glu was purified using cell fractionation, ultrafiltration, DEAE-and CM-cellulose chromatography columns. The purified CL-Glu was a monomer with a molecular weight of 70 +/- 3 kDa, isoelectric point of 7.2, optimum temperature of 70 degrees C and it was active over a broad pH range with a pH optimum at pH 8.0. Its activity had a clear dependence on phosphate ions. The studied enzyme showed sigmoidal kinetics, indicated its allosteric behavior with Km of 36 +/- 4 rnM and Hill coefficient of 1.5 which suggested a positive cooperatively of active sites. The purified L-glutaminase exerted antitumor activity against different cell lines with the highest cytotoxic activity against Hepatocellular carcinoma cell line (HepG-2) with an IC50 value of 152 mu g/ml. In conclusion, L-glutaminase was purified from camel liver using simple methods and its unique properties such as stability at both wide pH range and at high temperature along with its relatively low molecular weight, facilitated its usage in medical applications as antitumor drug. (C) 2019 Elsevier B.V. All rights reserved.
机译：L-谷氨酰胺酶在治疗各种癌症中的谷氨酰胺耗尽酶中获得了重要的关注。因此，本研究旨在纯化，表征和研究来自骆驼肝线粒体（CL-Glu）的抗肿瘤活性，因为没有关于来自骆驼的CL-Glu的可用信息。使用细胞分级，超滤，DEAE和CM-纤维素色谱柱纯化CL-Glu。将纯化的CL-Glu被的70 +/- 3kDa的，7.2，70℃下的最适温度的等电点的分子量的单体和它在广泛的pH范围内具有最适pH在pH8.0是活跃。其活性明显依赖磷酸盐离子。所研究的酶显示Sigmoidal动力学，表明其具有36 +/- 4 RNM和Hill系数的崩解行为，1.5的Hill系数，这表明了活性位点的正面合作。纯化的L-谷氨酰胺酶施加抗肿瘤活性对不同细胞系的抗肿瘤活性，其具有对肝细胞癌细胞系（HepG-2）的最高细胞毒性活性，IC50值为152μg/ ml。总之，使用简单的方法从骆驼肝纯化L-谷氨酰胺酶，其独特的性质如宽pH范围和高温下的稳定性以及其相对低的分子量，促进其在医疗应用中作为抗肿瘤药物的用途。 （c）2019 Elsevier B.v.保留所有权利。

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