使用双水相体系从平菇中同时提取和纯化多糖和蛋白质。
Simultaneous extraction and purification of polysaccharides and proteins from Pleurotus ostreatus using an aqueous two-phase system.
作者信息
Yang Zhilai, Wang Jingkun, Wang Hongyu, Xie Huixian, Zhang Yingnan, Wang Meng
机构信息
College of Food Science and Engineering, Nanjing University of Finance and Economics/Collaborative Innovation Center for Modern Grain Circulation and Safety, Nanjing, China.
Division of Pharmacoengineering and Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, North Carolina, USA.
出版信息
J Food Sci. 2025 Jan;90(1):e17674. doi: 10.1111/1750-3841.17674.
Pleurotus ostreatus is a nutrient-dense edible fungus renowned for its delicate texture, appealing flavor, and numerous potential health benefits. Simultaneous extraction within the framework of food resource processing facilitates the concurrent isolation and analysis of multiple target compounds. In this study, an ethanol/salt aqueous two-phase system (ATPS) was employed to extract polysaccharides (PS) and proteins from P. ostreatus. The impacts of pH, inorganic salts, and temperature on ATPS phase formation were systematically evaluated. The ethanol/K₂HPO₄ system demonstrated superior selective extraction performance under optimal conditions: 25.16% ethanol (w/w), 15.91% K₂HPO₄ (w/w), and 24.11% crude extract (w/w) without pH adjustment. The highest recovery efficiency of PS and proteins were 90.29% ± 0.24% and 76.35% ± 0.15%, respectively. The simultaneous extraction efficiency of PS and proteins was 12.49% ± 0.14% and 20.34% ± 0.09%, respectively. As a comparison, hot water extraction and alkali extraction methods were performed to assess the impact of ATPS on the physicochemical properties of PS and proteins. SDS-PAGE and HPLC analyses confirmed that the protein subunit distribution was consistent across different extraction methods, whereas the ATPS-extracted PS exhibited characteristics of homogeneous heteropolysaccharides with a molecular weight of 28.8 × 10⁶ Da. Monosaccharides such as mannose, glucuronic acid, and glucose were identified in the PS hydrolysate. The results demonstrate that ATPS preserves the physicochemical integrity of the extracted substances. This method holds promise for unlocking new possibilities and has the potential to become an effective method for large-scale extraction and purification of polysaccharides and proteins from edible fungi. PRACTICAL APPLICATION: This work describes an efficient and straightforward aqueous two-phase system (ATPS) method for simultaneously extracting polysaccharides and proteins from Pleurotus ostreatus. This approach offers advantages such as reduced operational time, lower toxicity solvents, and minimized byproduct waste for the extraction of bioactive substances from edible fungi. Additionally, the reduced residue levels help to shorten subsequent purification steps. This promising method has potential applications in the food, pharmaceutical, and health product industries.
平菇是一种营养丰富的食用菌，以其细腻的质地、诱人的风味和众多潜在的健康益处而闻名。在食品资源加工框架内进行同时提取，有助于同时分离和分析多种目标化合物。在本研究中，采用乙醇/盐水双水相体系（ATPS）从平菇中提取多糖（PS）和蛋白质。系统评估了pH值、无机盐和温度对ATPS相形成的影响。乙醇/K₂HPO₄体系在最佳条件下表现出优异的选择性提取性能：25.16%乙醇（w/w）、15.91%K₂HPO₄（w/w）和24.11%粗提取物（w/w），无需调节pH值。PS和蛋白质的最高回收率分别为90.29%±0.24%和76.35%±0.15%。PS和蛋白质的同时提取效率分别为12.49%±0.14%和20.34%±0.09%。作为比较，进行了热水提取和碱提取方法，以评估ATPS对PS和蛋白质理化性质的影响。SDS-PAGE和HPLC分析证实，不同提取方法的蛋白质亚基分布一致，而ATPS提取的PS表现出均一杂多糖的特征，分子量为28.8×10⁶Da。在PS水解产物中鉴定出甘露糖、葡萄糖醛酸和葡萄糖等单糖。结果表明，ATPS保留了提取物的理化完整性。该方法有望开辟新的可能性，有潜力成为从食用菌中大规模提取和纯化多糖和蛋白质的有效方法。实际应用：本工作描述了一种高效、简便的双水相体系（ATPS）方法，用于同时从平菇中提取多糖和蛋白质。这种方法具有操作时间短、溶剂毒性低、副产物浪费少等优点，可用于从食用菌中提取生物活性物质。此外，残留水平的降低有助于缩短后续的纯化步骤。这种有前景的方法在食品、制药和保健品行业具有潜在应用。
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