乙醇洗脱制备脱铁血红蛋白及其运载特性研究  

Study on preparation of apohemoglobin through ethanol elution and its complexation characteristics

作　　者：闵怡宁 朱思嘉 周成 张炎云 丁双 陈星 张娜娜 闫博文[1] 范大明[1] MIN Yining;ZHU Sijia;ZHOU Cheng;ZHANG Yanyun;DING Shuang;CHEN Xing;ZHANG Nana;YAN Bowen;FAN Daming(School of Food Science and Technology,Jiangnan University,Wuxi 214122,China)

机构地区：[1]江南大学食品学院,江苏无锡214122

出　　处：《食品与发酵工业》2025年第6期200-207,共8页Food and Fermentation Industries

基　　金：中国博士后科学基金面上项目(2023M731336);江南大学大学生创新创业训练计划资助(CXY2023009)。

摘　　要：畜禽血液有良好的营养价值与功能特性,高值化利用能提高产品附加值、拓宽应用前景,有利于推进生态环境保护和可持续发展。血液蛋白质中主要组分血红蛋白,其多肽链在非极性氨基酸残基富集的疏水区内可缔合血红素,脱除血红素生成脱铁血红蛋白。该研究采用乙醇洗脱法,使用不同体积分数的酸性乙醇洗脱血红素,计算脱铁血红蛋白的得率,对其结构、功能及重组特性进行分析。研究发现,乙醇体积分数80%的溶液透析脱除血红素的效果最好,经SDS-PAGE分析得脱铁血红蛋白纯度约为68%。圆二色光谱结果显示,脱铁血红蛋白α-螺旋含量降低,不同乙醇体积分数下二级结构变化趋势相近。重组血红蛋白的重组率约为40%。在姜黄素复合贮藏实验中,发现脱铁血红蛋白-姜黄素复合物的光和热稳定性较好,低温、弱光照强度有利于其稳定性。综上,采用乙醇洗脱法制备脱铁血红蛋白具有一定的合理性,既降低安全风险和生产成本又有较好的提取效果,可为禽畜血液的高值化利用提供参考。Livestock blood has good nutritional value and functional characteristics.The high-value utilization of livestock blood resources can enhance product value,broaden application prospects,and contribute to the protection of the ecological environment.This holds significant potential for China’s economic,social,and ecological sustainability.Hemoglobin,a key blood protein component,has a polypeptide chain that binds to heme in a hydrophobic region rich in non-polar amino acid residues,removing heme and yielding apohemoglobin.To remove heme,the study employed the ethanol elution method with varying concentrations of acidic ethanol.Subsequently,the study determined the yield of apohemoglobin and conducted a comparative analysis of its structure,function,and recombination characteristics.The study found that the optimal ethanol concentration for maximum heme removal was 80%.The purity of apohemoglobin was about 68%by SDS-PAGE.The circular dichroism spectrum showed a significant reduction in theα-helix content of apohemoglobin during acidic ethanol dissolution,and the secondary structure changed similarly under different ethanol concentrations.The recombination rate of heme and apohemoglobin to form recombinant hemoglobin was about 40%.During the composite storage experiment with curcumin,it was found that the light stability and thermal stability of the ApoHb-Cur complex were enhanced compared with single Cur.Specifically,lower temperature and weaker light intensities further enhanced the stability of the ApoHb-Cur complex.In conclusion,the ethanol elution method for preparing apohemoglobin was both practical and effective,reducing safety risks and production costs while maintaining a good extraction outcome.This approach provides valuable insights for the deep processing and high-value utilization of livestock blood resources.

关 键 词：脱铁血红蛋白 蛋白活性 结构特性 姜黄素 运载特性 

分 类 号：TS251.93[轻工技术与工程—农产品加工及贮藏工程]