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从药用真菌树舌灵芝(灵芝科)中分离并鉴定凝集素及其对HT-29结肠癌细胞增殖抑制活性的评估

Isolation and Characterization of Lectin from the Artist's Conk Medicinal Mushroom, Ganoderma applanatum (Agaricomycetes), and Evaluation of Its Antiproliferative Activity in HT-29 Colon Cancer Cells.

作者信息

Kumaran Sekar, Pandurangan Ashok Kumar, Shenbhagaraman Ramalingam, Esa Norhaizan Mohd

机构信息

Institute of Systems Biology, Universiti Kebangsaan Malaysia, Bangi, Selangor, Malaysia; Muthayammal Center for Advanced Research, Muthayammal College of Arts & Science, Rasipuram, Tamil Nadu, India; S&R Agro Bio-Tech, Chennai, Tamil Nadu, India; Department of Botany, Periyar University Constituent Arts and Science College, Harur, Tamil Nadu, India.

Muthayammal Center for Advanced Research, Muthayammal College of Arts & Science, Rasipuram, Tamil Nadu, India; Department of Nutrition and Dietetics, Faculty of Medicine and Health Sciences, Universiti of Putra Malaya, Serdang, Selangor, Malaysia.

出版信息

Int J Med Mushrooms. 2017;19(8):675-684. doi: 10.1615/IntJMedMushrooms.2017021274.

Abstract

The growth and lectin production of Ganoderma applanatum, a white rot fungus, was optimized in broth cultures. The fungus was found to have a higher growth rate and higher lectin activity when grown in a medium adjusted to pH 6.5 at 26°C under stationary conditions. Expression of lectin activity started in 5-day-old mycelial culture; maximum activity was expressed after the 15th day of incubation. Among the various carbon and nitrogen sources tested, the carbon source sucrose and the nitrogen source yeast extract support maximum growth and lectin production. Lectin from G. applanatum was purified by ammonium sulfate precipitation and ion exchange chromatography. The purified fraction revealed a single band with a molecular weight of 35.0 kDa. Moreover, carbohydrates such as mannitol, glucose, sucrose, maltose, mannose, galactose, sorbose, and fructose were found to inhibit the hemagglutinating activity of the lectin. The purified lectins from G. applanatum contain cytotoxic and proapoptotic activities against HT-29 colon adenocarcinoma cells.

摘要

在液体培养中对一种白腐真菌——平盖灵芝的生长和凝集素生产进行了优化。发现该真菌在26°C、静止条件下于pH 6.5的培养基中生长时,具有更高的生长速率和更高的凝集素活性。凝集素活性在5日龄菌丝体培养物中开始表达;培养15天后表达出最大活性。在所测试的各种碳源和氮源中,碳源蔗糖和氮源酵母提取物支持最大生长和凝集素生产。通过硫酸铵沉淀和离子交换色谱法纯化了平盖灵芝的凝集素。纯化后的组分显示出一条分子量为35.0 kDa的单一条带。此外,发现甘露醇、葡萄糖、蔗糖、麦芽糖甘露糖、半乳糖、山梨糖和果糖等碳水化合物会抑制该凝集素的血凝活性。从平盖灵芝中纯化得到的凝集素对HT - 29结肠腺癌细胞具有细胞毒性和促凋亡活性。

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