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首页> 外文期刊>The Journal of Antibiotics: An International Journal >Production of Ansamycin Polyketide Precursors in Escherichia coli
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Production of Ansamycin Polyketide Precursors in Escherichia coli

机译：在大肠杆菌中生产安沙霉素聚酮化合物前体

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For the heterologous production of ansamycin polyketides such as rifamycin and geldanamycin in Escherichia coli,a number of unusual but important tools must be engineered into the bacterium.Here we demonstrate efficient production of the starter unit 3-amino-5-hydroxybenzoic acid (AHBA) and the methoxymalonyl extender unit in E.coli.Previous work has demonstrated the production of the ansamycin starter unit AHBA in E.coli in low yield.It was shown that the low yield is primarily due to acetylation of AHBA into N-acetyl-AHBA.Three methods for minimizing this side reaction were evaluated.First,a putative N-arylamine-acetyltransferase (NAT) was deleted from the E.coli chromosome,although this did not alter N-acetyl-AHBA production.Next,E.coli grown in media devoid of glucose yielded a nearly equal mixture of AHBA and N-acetyl-AHBA.Lastly,the NAT inhibitor glycyrrhizic acid was shown to further inhibit the acetylation reaction.The entire set of genes for synthesizing the methoxymalonyl extender unit was transferred from the geldanamycin producer Streptomyces hygroscopicus into E.coli.The pathway specific ACP isolated from the resulting recombinant strain was found to predominantly occur as methyoxymalonyl-ACP.Together,these findings set the stage for engineered biosynthesis of ansamycin polyketides in E.coli.
机译：为了在大肠杆菌中异源生产安沙霉素聚酮化合物（如利福霉素和格尔德霉素），必须在细菌中设计许多不寻常但重要的工具。在此，我们证明了高效生产3-氨基-5-羟基苯甲酸起始剂单元的方法。以前的工作证明了在大肠杆菌中以低收率生产安沙霉素起始剂单元AHBA，这表明低收率主要是由于AHBA乙酰化为N-乙酰基AHBA评估了最小化这种副反应的三种方法。首先，从大肠杆菌染色体中删除了一个假定的N-芳基胺-乙酰基转移酶（NAT），尽管这不会改变N-乙酰基-AHBA的产生。在没有葡萄糖的培养基中，AHBA和N-乙酰基-AHBA的混合物几乎相等。最后，NAT抑制剂甘草酸被证明进一步抑制了乙酰化反应。整个合成甲氧基丙二酸前体的基因将ender单位从格尔德霉素生产商吸水链霉菌转移到大肠杆菌中。发现从重组菌株中分离出的途径特异性ACP主要以甲氧基丙二酰-ACP的形式存在。这些发现共同为在大肠杆菌中工程合成生物合成安沙霉素聚酮化合物奠定了基础。 .coli。

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