Improving heterologous production of phenylpropanoids in Saccharomyces cerevisiae by tackling an unwanted side reaction of Tsc13, an endogenous double-bond reductase.
Improving heterologous production of phenylpropanoids in Saccharomyces cerevisiae by tackling an unwanted side reaction of Tsc13, an endogenous double-bond reductase.
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DOI：
10.1093/femsyr/fox004
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发表时间：
2017-01-01
影响因子：
3.2
通讯作者：
Naesby M
中科院分区：
文献类型：
--
作者：
Lehka BJ;Eichenberger M;Bjørn-Yoshimoto WE;Vanegas KG;Buijs N;Jensen NB;Dyekjær JD;Jenssen H;Simon E;Naesby M
苯丙类化合物，如黄酮类化合物和苯乙烯类化合物，具有很大的商业价值，在酿酒酵母中生产它们是一个很有前途的策略。然而，为了实现商业上可行的生产，过程的每一步都必须优化。我们观察了碳损失，已知发生在酵母的异源类黄酮途径中，并确定了一种内源性酶，烯丙基还原酶Tsc13，该酶通过还原对coumaryl - coa来积累间绿酸。Tsc13是参与脂肪酸合成的必需酶，不能被删除。因此，采用了两种方法，试图在不破坏自然功能的情况下降低副活性：位点饱和诱变确定了一些氨基酸的变化，这些变化略微增加了类黄酮的产生，但没有减少副产物的形成。相反，TSC13与植物基因同源物的互补基本上消除了不必要的副反应，同时在模拟补料分批发酵中保持了苯丙素的产量。用来自苹果的相应基因替代酵母还原酶，通过消除副产品的形成，可以更有效地生产黄酮类化合物。
Phenylpropanoids, such as flavonoids and stilbenoids, are of great commercial interest, and their production in Saccharomyces cerevisiae is a very promising strategy. However, to achieve commercially viable production, each step of the process must be optimised. We looked at carbon loss, known to occur in the heterologous flavonoid pathway in yeast, and identified an endogenous enzyme, the enoyl reductase Tsc13, which turned out to be responsible for the accumulation of phloretic acid via reduction of p-coumaroyl-CoA. Tsc13 is an essential enzyme involved in fatty acid synthesis and cannot be deleted. Hence, two approaches were adopted in an attempt to reduce the side activity without disrupting the natural function: site saturation mutagenesis identified a number of amino acid changes which slightly increased flavonoid production but without reducing the formation of the side product. Conversely, the complementation of TSC13 by a plant gene homologue essentially eliminated the unwanted side reaction, while retaining the productivity of phenylpropanoids in a simulated fed batch fermentation. Replacement of essential yeast reductase with corresponding gene from apple allows more efficient production of flavonoids by eliminating formation of side products.
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影响因子：
14.8
作者：
Gietz, R. Daniel;Schiestl, Robert H.
通讯作者：
Schiestl, Robert H.
影响因子：
6.4
作者：
Kuijpers NG;Solis-Escalante D;Bosman L;van den Broek M;Pronk JT;Daran JM;Daran-Lapujade P
通讯作者：
Daran-Lapujade P
影响因子：
6.4
作者：
Kildegaard KR;Jensen NB;Schneider K;Czarnotta E;Özdemir E;Klein T;Maury J;Ebert BE;Christensen HB;Chen Y;Kim IK;Herrgård MJ;Blank LM;Forster J;Nielsen J;Borodina I
通讯作者：
Borodina I
酿酒酵母的酿酒酵母的代谢工程从从头生产二氢切尔科酮，具有已知的抗氧化剂，抗糖尿病和甜味特性。
影响因子：
8.4
作者：
Eichenberger M;Lehka BJ;Folly C;Fischer D;Martens S;Simón E;Naesby M
通讯作者：
Naesby M
影响因子：
3.2
作者：
Jensen NB;Strucko T;Kildegaard KR;David F;Maury J;Mortensen UH;Forster J;Nielsen J;Borodina I
通讯作者：
Borodina I