Simple Purification of Nicotiana benthamiana-Produced Recombinant Colicins: High-Yield Recovery of Purified Proteins with Minimum Alkaloid Content Supports the Suitability of the Host for Manufacturing Food Additives.
Simple Purification of Nicotiana benthamiana-Produced Recombinant Colicins: High-Yield Recovery of Purified Proteins with Minimum Alkaloid Content Supports the Suitability of the Host for Manufacturing Food Additives.
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DOI：
10.3390/ijms19010095
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发表时间：
2017-12-29
影响因子：
5.6
通讯作者：
Gleba Y
中科院分区：
文献类型：
--
作者：
Stephan A;Hahn-Löbmann S;Rosche F;Buchholz M;Giritch A;Gleba Y
大肠杆菌素是天然的非抗生素细菌蛋白质，具有窄谱，但具有极高的抗菌活性。这些蛋白是控制主要致病性产滋贺毒素大肠杆菌的有希望的食品添加剂。肉类和农产品中的大肠杆菌血清型。在美国，从菠菜和叶甜菜等可食用植物中产生的大肠杆菌素已经被美国食品和药物管理局接受。S.美国食品和药物管理局（FDA）和美国。S.美国农业部（USDA）通过GRAS（公认安全）监管审查程序将其列为食品加工抗菌剂。本氏烟草（Nicotianabenthamiana）是烟草的一种野生近缘种; tabacum已经成为生产重组蛋白质（包括生物药物、疫苗和生物材料）的优选生产宿主植物，但是迄今为止所采用的纯化方法非常复杂和昂贵。我们描述了一种简单而廉价的纯化方法的基础上特定的酸性提取，然后通过一个色谱步骤。该方法提供了纯化的大肠杆菌素的高回收率，以及尼古丁的急剧减少到能够使最终产品用于食品的水平。所描述的纯化方法允许以商业上可行的商品成本生产大肠杆菌素产品，并且可能广泛适用于其他成本敏感的蛋白质。
Colicins are natural non-antibiotic bacterial proteins with a narrow spectrum but an extremely high antibacterial activity. These proteins are promising food additives for the control of major pathogenic Shiga toxin-producing E. coli serovars in meats and produce. In the USA, colicins produced in edible plants such as spinach and leafy beets have already been accepted by the U. S. Food and Drug Administration (FDA) and U. S. Department of Agriculture (USDA) as food-processing antibacterials through the GRAS (generally recognized as safe) regulatory review process. Nicotiana benthamiana, a wild relative of tobacco, N. tabacum, has become the preferred production host plant for manufacturing recombinant proteins—including biopharmaceuticals, vaccines, and biomaterials—but the purification procedures that have been employed thus far are highly complex and costly. We describe a simple and inexpensive purification method based on specific acidic extraction followed by one chromatography step. The method provides for a high recovery yield of purified colicins, as well as a drastic reduction of nicotine to levels that could enable the final products to be used on food. The described purification method allows production of the colicin products at a commercially viable cost of goods and might be broadly applicable to other cost-sensitive proteins.
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影响因子：
5.5
作者：
Cummings JF;Guerrero ML;Moon JE;Waterman P;Nielsen RK;Jefferson S;Gross FL;Hancock K;Katz JM;Yusibov V;Fraunhofer USA Center for Molecular Biotechnology Study Group
通讯作者：
Fraunhofer USA Center for Molecular Biotechnology Study Group
影响因子：
50.5
作者：
Bendandi, M.;Marillonnet, S.;Gleba, Y.
通讯作者：
Gleba, Y.
影响因子：
3.8
作者：
SAITOH, F;NOMA, M;KAWASHIMA, N
通讯作者：
KAWASHIMA, N
DOI：
10.1515/cttr-2016-0008
发表时间：
2016-04-01
期刊：
Beitraege zur Tabakforschung International
影响因子：
--
作者：
Moldoveanu, Serban C.;Scott, Wayne A.;Lawson, Darlene M.
通讯作者：
Lawson, Darlene M.
影响因子：
64.8
作者：
Qiu, Xiangguo;Wong, Gary;Audet, Jonathan;Bello, Alexander;Fernando, Lisa;Alimonti, Judie B.;Fausther-Bovendo, Hugues;Wei, Haiyan;Aviles, Jenna;Hiatt, Ernie;Johnson, Ashley;Morton, Josh;Swope, Kelsi;Bohorov, Ognian;Bohorova, Natasha;Goodman, Charles;Kim, Do;Pauly, Michael H.;Velasco, Jesus;Pettitt, James;Olinger, Gene G.;Whaley, Kevin;Xu, Bianli;Strong, James E.;Zeitlin, Larry;Kobinger, Gary P.
通讯作者：
Kobinger, Gary P.