[1]陶梦娜,潘月明,涂璐璐,等.丙酮-低共熔溶剂联合提取法的构建及其在蛋白质提取中的应用[J].江苏农业学报,2025,(12):2326-2333.[doi:doi:10.3969/j.issn.1000-4440.2025.11.005]
　TAO Mengna,PAN Yueming,TU Lulu,et al.Construction of acetone-deep eutectic solvent combined extraction method and its application in protein extraction[J].,2025,(12):2326-2333.[doi:doi:10.3969/j.issn.1000-4440.2025.11.005]
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丙酮-低共熔溶剂联合提取法的构建及其在蛋白质提取中的应用()

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2025年12期
页码:
2326-2333
栏目:
遗传育种·生理生化
出版日期:
2025-12-31

文章信息/Info

Title:
Construction of acetone-deep eutectic solvent combined extraction method and its application in protein extraction
作者:
陶梦娜1; 2; 潘月明1; 2; 涂璐璐1; 2; 王传洗1; 2; 王震宇1; 2
(1.江南大学环境与生态学院,无锡江苏214122；2.江南大学环境过程与污染控制研究所,无锡江苏214122)
Author(s):
TAO Mengna1; 2; PAN Yueming1; 2; TU Lulu1; 2; WANG Chuanxi1; 2; WANG Zhenyu1; 2
(1.School of Environment and Ecology, Jiangnan University, Wuxi 214122, China；2.Institute of Environmental Processes and Pollution Control, Jiangnan University, Wuxi 214122, China)
关键词:
蛋白质提取; 丙酮-低共熔溶剂联合提取法; 液相色谱-串联质谱; 植物蛋白质组学
Keywords:
protein extraction; acetone-deep eutectic solvent combined extraction method; liquid chromatography-tandem mass spectrometry; plant proteomics
分类号:
S565.1
DOI:
doi:10.3969/j.issn.1000-4440.2025.11.005
文献标志码:
A
摘要:
植物蛋白质组学在阐明作物生长发育进程、解析品质改良分子机制及揭示作物抗逆应答通路等方面具有重要意义。然而,传统的三氯乙酸-丙酮沉淀法对疏水性蛋白、膜结合蛋白及热稳定性蛋白提取效率较低,而凝胶电泳-质谱联用技术对高相对分子量、低相对分子量和极碱性蛋白的检测能力有限,导致蛋白质提取率低、种类覆盖度不足。本研究利用低共熔溶剂(DES)具有绿色低毒、溶解能力强、可高效提取疏水性及热稳定性蛋白等优势,建立了丙酮-DES联合提取法。该方法显著提升了大豆种子蛋白的提取效率和鉴定数目,蛋白质提取率提高至28.9%,共鉴定到2 665种蛋白质,包括参与细胞物质运输与抗氧化过程的内质网跨膜蛋白和过氧化物酶体膜蛋白PEX14,与低氮胁迫耐受性相关的含CBS结构域的蛋白质,以及作为植物铁营养来源的铁蛋白等。利用丙酮-DES联合提取法提取玉米、花生、芝麻的蛋白质,与传统方法相比,丙酮-DES联合提取法操作时间更短,在蛋白质提取率、种类覆盖度和重复性方面均具有显著优势,且对不同蛋白质含量、不同含油量的种子原料均展现出良好的适用性。本研究结果为作物功能基因挖掘及品质改良提供了技术支持。
Abstract:
Plant proteomics plays a vital role in elucidating the growth and development processes of crops, deciphering the molecular mechanism underlying quality improvement, and revealing the stress-responsive pathways of crops. However, the traditional trichloroacetic acid-acetone precipitation method exhibits low extraction efficiency for hydrophobic proteins, membrane-bound proteins and thermostable proteins. Meanwhile, the gel electrophoresis-mass spectrometry technology has limited detection capability for high relative molecular weight, low relative molecular weight and extremely alkaline proteins, which results in low protein extraction rate and insufficient species coverage. Taking advantage of deep eutectic solvents (DES) featuring green and low toxicity, strong dissolving capacity, and high-efficiency extraction of hydrophobic and thermostable proteins, this study established an acetone-DES combined extraction method. This method significantly improved the extraction efficiency and identification number of soybean seed proteins, the protein extraction rate was increased to 28.9%, and a total of 2 665 proteins were identified. These identified proteins included endoplasmic reticulum transmembrane proteins and peroxisomal membrane protein PEX14 involved in cellular material transport and antioxidant processes, CBS domain-containing proteins associated with low-nitrogen stress tolerance, as well as ferritin serving as a source of plant iron nutrition. When applied to extract proteins from corn, peanut and sesame, compared with traditional methods, the acetone-DES combined extraction method showed shorter operation time, and exhibited significant advantages in protein extraction rate, species coverage and reproducibility. Moreover, it demonstrated good applicability to seed materials with different protein contents and different oil contents. This study provides technical support for the mining of crop functional genes and crop quality improvement.

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备注/Memo

备注/Memo:
收稿日期:2025-10-14基金项目:国家自然科学基金青年基金项目(42407609)；无锡市科技发展资金项目“太湖之光”科技攻关基础研究项目(K20231031)作者简介:陶梦娜(1993-),女,江苏无锡人,博士,实验师,研究方向为植物蛋白质组学、人工纳米颗粒的环境地球化学过程。(E-mail)taomengna@jiangnan.edu.cn通讯作者:王传洗,(E-mail)wangcx2018@jiangnan.edu.cn
更新日期/Last Update: 2026-01-20