江苏农业学报2025，Vol.41Issue(12)：2326-2333,8.DOI:10.3969/j.issn.1000-4440.2025.12.005
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
Construction of acetone-deep eutectic solvent combined extraction method and its application in protein extraction
摘要
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 meth-od.This method significantly improved the extraction efficien-cy 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 in-cluded 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 repro-ducibility.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.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Key words
protein extraction/acetone-deep eutectic solvent combined extraction method/liquid chromatography-tandem mass spectrometry/plant proteomics分类
农业科技引用本文复制引用
陶梦娜,潘月明,涂璐璐,王传洗,王震宇..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,2025,41(12):2326-2333,8.基金项目
国家自然科学基金青年基金项目(42407609) （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) （K20231031）
