Production of pyranoanthocyanins using Escherichia coli co-cultures
Production of pyranoanthocyanins using Escherichia coli co-cultures
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DOI：
10.1016/j.ymben.2019.05.008
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发表时间：
2019-09-01
影响因子：
8.4
通讯作者：
Koffas, Mattheos A. G.
中科院分区：
文献类型：
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作者：
Akdemir, Hulya;Silva, Adilson;Koffas, Mattheos A. G.
羟基苯基-吡喃花青素是红葡萄酒和一些果汁中发现的吡喃花青素之一。由于它们有一个四环（吡喃或环D），与花青素前体相比，它提供了更高的颜色强度和对pH变化的卓越稳定性，这些分子是最重要的天然着色剂之一，特别是在低酸性和中酸性食品和饮料中。然而，由于它们的浓度很低，分离和表征是困难的。在这项研究中，我们共培养重组大肠杆菌菌株，以提高滴度和产量合成pyrano花青素。为了完成这项任务，我们首先设计了4-乙烯基苯酚和4-乙烯基儿茶酚生产模块，然后将这些菌株与花青素-3- o -葡萄糖苷生产重组细胞共培养，得到pyranocyanidin-3- o -葡萄糖苷苯酚（花青素-3- o -葡萄糖苷与乙烯基儿茶酚加合物）和pyranocyanidin-3- o -葡萄糖苷-儿茶酚（花青素-3- o -葡萄糖苷与乙烯基儿茶酚加合物）。通过优化共培养条件，我们能够显著提高最终滴度和产量，使我们的共培养方法轻松优于从红酒中生产吡喃花青素。最后，我们证明了生产的pyrano花青素远比起始植物生产的花青素3- o -葡萄糖苷更稳定。
Hydroxyphenyl-pyranoanthocyanins are one of the pyranoanthocyanins found in red wines and some fruit juices. Since they have a fourth ring (pyran or ring D) which provides higher color intensity and exceptional stability toward pH variations in comparison to their anthocyanin precursors, these molecules are one of the most important candidates as natural colorants especially for low- and medium-acidic food and beverages. However, their isolation and characterization are difficult due to their very low concentration. In this study, we co-cultured recombinant E. coli strains to synthesize pyranoanthocyanins with improved titers and yields. To accomplish this task, firstly we engineered 4-vinylphenol and 4-vinylcatechol producer modules then we cocultured each one of these strains with cyanidin-3-O-glucoside producer recombinant cells to obtain pyranocyanidin-3-O-glucoside phenol (cyanidin-3-O-glucoside with vinylphenol adduct) and pyranocyanidin-3-O-glucoside-catechol (cyanidin-3-O-glucoside with vinylcatechol adduct). By optimizing the co-culture conditions, we were able to significantly increase final titers and yields, allowing our co-culture approach to easily outperform production of pyranoanthocyanins from red wine. Finally, we demonstrate that the produced pyranoanthocyanins are far more stable than the starting plant-produced cyanidin 3-O-glucoside.