凡纳对虾原肌球蛋白硫酸铵沉淀分离纯化方法的优化

doi:10.7506/spkx1002-6630-201718030

食品科学 ›› 2017, Vol. 38 ›› Issue (18): 187-192.doi: 10.7506/spkx1002-6630-201718030

凡纳对虾原肌球蛋白硫酸铵沉淀分离纯化方法的优化

傅玲琳，富舒洁，王彦波，王翀，张岩，吴琦，马爱进

（1.浙江工商大学食品与生物工程学院，浙江?杭州 310018；2.河北省食品检测研究院，河北?石家庄 050091；3.中国标准化研究院，北京 100191）

出版日期:2017-09-25 发布日期:2017-09-04
基金资助:
国家自然科学基金面上项目（31571770）；“十三五”国家重点研发计划重点专项（2016YFD0401203）；浙江省科技厅公益技术应用研究计划项目（2016C32058）

Optimization of Purification by Ammonium Sulfate Precipitation of the Major Allergen Tropomyosin from Shrimp (Litopenaeus vannamei)

FU Linglin, FU Shujie, WANG Yanbo, WANG Chong, ZHANG Yan, WU Qi, MA Aijin

(1. School of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou 310018, China; 2. Hebei Food Inspection and Research Institute, Shijiazhuang 050091, China; 3. China National Institute of Standardization, Beijing 100191, China)

Online:2017-09-25 Published:2017-09-04

摘要/Abstract

摘要： 虾原肌球蛋白（tropomyosin，TM）食物过敏是一种全球性的食品安全问题，研究发现TM的分离纯化对于系统准确地鉴定和控制过敏具有重要的意义，鉴于此，本研究以南美白对虾（Litopenaeus vannamei）为对象，改进了TM分离纯化的方法。结果表明，抽提液pH?6.5～7.5、硫酸铵饱和度30%时，可以有效分离纯化TM；经喹啉酸法测定其质量浓度最高可达43?μg/μL。该方法省去了高效液相色谱、层析技术等对TM的进一步纯化，减少了纯化步骤，避免了在这些纯化步骤中出现的蛋白损失及蛋白稀释；高质量浓度的TM溶液可无须经过冷冻干燥，直接用于对其理化性质及过敏原性的研究。

关键词: 过敏原, 南美白对虾, 原肌球蛋白, 分离, 纯化

Abstract: Tropomyosin (TM), the major shrimp allergen, is one of the serious food safety problems throughout the world and has drawn much attention. It has been reported that the isolation and purification of TM is of very importance in the identification and control of allergy. In the present study, tropomyosin was purified from Litopenaeus vannamei by an improved method. The results indicated that TM could be purified effectively by adjusting the extraction buffer to pH 6.5 to 7.5 and then adding ammonium sulfate to approximately 30% saturation. The concentration of TM was about 43 μg/μL, as determined by a BCA Protein Assay Kit. The improved method avoided further purification of TM using high performance liquid chromatography (HPLC) or another chromatographic technology. In addition, the process of TM isolation was simplified and TM loss and dilution were avoided during further purification. High concentration of purified TM solution obtained in the present study could be directly used to study its physicochemical properties and allergenicity without being freeze-dried.

Key words: allergen, Litopenaeus vannamei, tropomyosin (TM), isolation, purification

中图分类号:

TS254.1

傅玲琳，富舒洁，王彦波，王翀，张岩，吴琦，马爱进. 凡纳对虾原肌球蛋白硫酸铵沉淀分离纯化方法的优化[J]. 食品科学, 2017, 38(18): 187-192.

FU Linglin, FU Shujie, WANG Yanbo, WANG Chong, ZHANG Yan, WU Qi, MA Aijin. Optimization of Purification by Ammonium Sulfate Precipitation of the Major Allergen Tropomyosin from Shrimp (Litopenaeus vannamei)[J]. FOOD SCIENCE, 2017, 38(18): 187-192.

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凡纳对虾原肌球蛋白硫酸铵沉淀分离纯化方法的优化

Optimization of Purification by Ammonium Sulfate Precipitation of the Major Allergen Tropomyosin from Shrimp (Litopenaeus vannamei)

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