文章基本信息

标题：Functional expression of the sweet-tasting protein brazzein in transgenic tobacco
本地全文：下载
作者：CHOI Hyo-Eun ; LEE Ji-In ; JO Seon-Yeong 等
期刊名称：Food Science and Technology (Campinas)
印刷版ISSN：0101-2061
电子版ISSN：1678-457X
出版年度：2022
卷号：42
DOI：10.1590/fst.40521
语种：English
出版社：Sociedade Brasileira de Ciência e Tecnologia de Alimentos
摘要：The sweet-tasting protein, brazzein, has potential as a low-calorie sugar substitute owing to its high sweetness, stability, and water solubility. In this study, the synthetic brazzein gene was expressed in the tobacco plant, Nicotiana tabacum. Three types of expression cassettes containing the brazzein gene were constructed to examine the expression and purification efficiency of the brazzein: pBI-BZ1 containing a signal sequence and His-tag, pBI-BZ2 containing a signal sequence, and pBI-BZ3 containing only the brazzein gene. Brazzein expression confirmed by ELISA was purified using ammonium sulfate precipitation, heat treatment, and CM-sepharose chromatography. The purity and conformational state of the brazzein were confirmed using SDS-PAGE, HPLC, and circular dichroism. The identity of the brazzein was confirmed by N-terminal amino acid analysis, ESI-MS/MS, and sweetness analysis. We successfully generated brazzein overexpression tobacco plants, suggesting that this method could be used as a brazzein production platform to provide an alternative to currently produced sweeteners.
关键词：alternative sweetener;brazzein;Agrobacterium;mediated transformation;transgenic tobacco plant;protein purification