Glucosinolate-Mediated Inhibition of Carbonic Anhydrase IX: A Study on Turnip (Brassica rapa) and its Potential in Cancer Prevention

IF 3.2 4区农林科学 Q2 FOOD SCIENCE & TECHNOLOGY

Food Biophysics Pub Date : 2025-06-27 DOI:10.1007/s11483-025-09992-5

Emadeldin M. Kamel, Ahmed A. Allam, Hassan A. Rudayni, Saleh Alkhedhairi, Faris F. Aba Alkhayl, May Bin-Jumah, Doaa A. Abdelrheem, Al Mokhtar Lamsabhi

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引用次数: 0

Abstract

Cruciferous vegetables such as turnip (Brassica rapa) are rich in bioactive glucosinolates with potential health-promoting and anticancer properties. Here, 4-methoxyglucobrassicin, glucobrassicin and neoglucobrassicin were purified from hot-methanol extracts of B. rapa roots by acidic-alumina and Sephadex LH-20 chromatography. Recombinant human carbonic anhydrase IX (CA IX) esterase activity was measured with a 4-nitrophenyl-acetate microplate assay, and inhibition constants were extracted from Lineweaver–Burk plots. Protein–ligand interactions were investigated by AutoDock Vina docking (PDB 5FL4), followed by 200 ns CHARMM36m molecular-dynamics simulations in GROMACS; binding free energies were obtained with the MM/PBSA method, and conformational stability was evaluated through free-energy-landscape analysis. In vitro enzyme-inhibition assays revealed that neoglucobrassicin and glucobrassicin exhibited potent CA IX inhibition, with IC₅₀ values of 48.5 ± 2.1 nM and 65.06 ± 2.8 nM, respectively. Kinetic studies indicated a mixed-type inhibition mechanism, suggesting multiple interaction modes with the enzyme. Molecular docking confirmed strong polar interactions within the CA IX active site, and MD/MM-PBSA ranked neoglucobrassicin as the most favorable binder (ΔG_total = − 34.74 ± 2.47 kcal mol⁻¹). Free-energy-landscape analysis further showed that neoglucobrassicin maintains a highly stable conformational state during the simulation. Overall, this integrated isolation, enzyme-assay, and computational workflow identifies turnip-derived indole glucosinolates as promising natural CA IX inhibitors and supports their potential role in functional foods and nutraceuticals aimed at cancer prevention and tumor-micro-environment modulation.

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硫代葡萄糖苷介导的碳酸酐酶IX的抑制作用：芜菁及其抗癌潜力的研究

十字花科蔬菜如萝卜（Brassica rapa）富含生物活性硫代葡萄糖苷，具有潜在的促进健康和抗癌特性。本研究采用酸性氧化铝和Sephadex LH-20层析，从rapa根热甲醇提取物中分离得到4-甲氧基葡萄花青素、葡萄花青素和新葡萄花青素。采用4-硝基苯乙酸微孔板法测定重组人碳酸酐酶IX （CA IX）酯酶活性，并从Lineweaver-Burk图中提取抑制常数。通过AutoDock Vina对接（PDB 5FL4）研究蛋白质与配体的相互作用，然后在GROMACS中进行200 ns CHARMM36m分子动力学模拟；用MM/PBSA法获得了结合自由能，并通过自由能景观分析评价了构象稳定性。体外酶抑制实验表明，新糖花椰菜素和葡萄花椰菜素对CA IX具有较强的抑制作用，IC50值分别为48.5±2.1 nM和65.06±2.8 nM。动力学研究表明其抑制机制为混合型，表明其与酶具有多种相互作用模式。分子对接证实了CA IX活性位点内的强极性相互作用，MD/MM-PBSA将新葡萄花青素列为最有利的结合剂（ΔGtotal =−34.74±2.47 kcal mol⁻¹）。自由能景观分析进一步表明，在模拟过程中，新葡萄糖花青素保持高度稳定的构象状态。总的来说，这种综合的分离、酶测定和计算工作流程确定了芜菁衍生的吲哚硫代葡萄糖苷是有前途的天然CA IX抑制剂，并支持其在功能性食品和营养保健品中的潜在作用，旨在预防癌症和调节肿瘤微环境。

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来源期刊

Food Biophysics 工程技术-食品科技

CiteScore

5.80

自引率

3.30%

发文量

审稿时长

1 months

期刊介绍： Biophysical studies of foods and agricultural products involve research at the interface of chemistry, biology, and engineering, as well as the new interdisciplinary areas of materials science and nanotechnology. Such studies include but are certainly not limited to research in the following areas: the structure of food molecules, biopolymers, and biomaterials on the molecular, microscopic, and mesoscopic scales; the molecular basis of structure generation and maintenance in specific foods, feeds, food processing operations, and agricultural products; the mechanisms of microbial growth, death and antimicrobial action; structure/function relationships in food and agricultural biopolymers; novel biophysical techniques (spectroscopic, microscopic, thermal, rheological, etc.) for structural and dynamical characterization of food and agricultural materials and products; the properties of amorphous biomaterials and their influence on chemical reaction rate, microbial growth, or sensory properties; and molecular mechanisms of taste and smell. A hallmark of such research is a dependence on various methods of instrumental analysis that provide information on the molecular level, on various physical and chemical theories used to understand the interrelations among biological molecules, and an attempt to relate macroscopic chemical and physical properties and biological functions to the molecular structure and microscopic organization of the biological material.