研究报告

饱和硫酸铵纯化重组日本囊对虾铁蛋白

  • 马贵红 ,
  • 赖龙昕 ,
  • 李蝶 ,
  • 冯颖琳 ,
  • 李筱 ,
  • 张志清 ,
  • 李树红 ,
  • 王彩霞 ,
  • 顾毅 ,
  • 马翼 ,
  • 李美良
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  • 1(四川农业大学 食品学院,四川 雅安,625014)
    2(绵阳钟沟生态农业科技有限公司,四川 绵阳,621000)
    3(四川润兆渔业有限公司,四川 雅安,625000)
硕士研究生(李美良副教授为通信作者,E-mail:liml@sicau.edu.cn)

收稿日期: 2021-09-08

  修回日期: 2021-10-11

  网络出版日期: 2022-09-16

基金资助

四川省科技厅项目(20ZDYF1893);四川省大学生创新创业训练计划项目(1921996121);四川农业大学科研兴趣项目(2021996689;1911113425)

Purification of recombinant Marsupenaeus japonicus ferritin by saturated ammonium sulfate

  • MA Guihong ,
  • LAI Longxin ,
  • LI Die ,
  • FENG Yinglin ,
  • LI Xiao ,
  • ZHANG Zhiqing ,
  • LI Shuhong ,
  • WANG Caixia ,
  • GU Yi ,
  • MA Yi ,
  • LI Meiliang
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  • 1(College of Food Science, Sichuan Agricultural University, Ya′an 625014, China)
    2(Mianyang Zhonggou Ecological Agriculture Technology Co.Ltd., Mianyang 621000, China)
    3(Sichuan Runzhao Fishery Co.Ltd., Ya′an 625000, China)

Received date: 2021-09-08

  Revised date: 2021-10-11

  Online published: 2022-09-16

摘要

目前,重组铁蛋白被广泛用作功能性纳米材料。该研究首先对热处理(65 ℃,10 min)后的重组日本囊对虾铁蛋白(recombinant Marsupenaeus japonicus ferritin,rMjF)粗提液采用20%饱和度的饱和硫酸铵处理,之后将其上清液中硫酸铵的饱和度提升至40%,并将其沉淀复溶于缓冲液后结合透析、超滤等非柱层析技术成功纯化了rMjF。结果表明,rMjF加热粗提液经饱和硫酸铵复合处理会大大降低杂蛋白的含量;rMjF是只含有1个亚基(约20 kDa)的脱铁铁蛋白,外径为12 nm左右;红外光谱分析显示,rMjF中ɑ-螺旋相对含量最高,为57.91%。此方法便于大批量分离纯化rMjF,可为rMjF作为纳米运载体的研究提供参考。

本文引用格式

马贵红 , 赖龙昕 , 李蝶 , 冯颖琳 , 李筱 , 张志清 , 李树红 , 王彩霞 , 顾毅 , 马翼 , 李美良 . 饱和硫酸铵纯化重组日本囊对虾铁蛋白[J]. 食品与发酵工业, 2022 , 48(16) : 138 -143 . DOI: 10.13995/j.cnki.11-1802/ts.029329

Abstract

At present, recombinant ferritin is widely used as functional nanomaterial. Firstly, after heat treatment (65 ℃, 10 min), the crude extract of recombinant Marsupenaeus japonicus ferritin (rMjF) was treated with 20% saturated ammonium sulfate. Secondly, the saturation of ammonium sulfate in the treated supernatant increased to 40%. Then, the precipitate was dissolved in buffer again. Finally, rMjF was successfully purified by non-column chromatography such as dialysis and ultrafiltration. The results showed that the crude extract of rMjF after heating treatment was treated with saturated ammonium sulfate compound treatment, the content of hybrid protein greatly reduced. The rMjF was apoferritin with only one subunit (about 20 kDa), with an outer diameter of about 12 nm. Fourier transform infrared spectroscopy analysis showed that the highest ɑ-helix relative content in rMjF was 57.91%. This method is convenient for mass separation and purification of rMjF, and provides a certain reference value for the study of rMjF as a nano transport carrier.

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