A BioBricks Metabolic Engineering Platform for the Biosynthesis of Anthracyclinones in Streptomyces coelicolor.
A BioBricks Metabolic Engineering Platform for the Biosynthesis of Anthracyclinones in Streptomyces coelicolor.
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DOI：
10.1021/acssynbio.2c00498
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发表时间：
2022-12-16
影响因子：
4.7
通讯作者：
Nybo, S. Eric
中科院分区：
文献类型：
--
作者：
Wang, Rongbin;Nguyen, Jennifer;Hecht, Jacob;Schwartz, Nora;Brown, Katelyn, V;Ponomareva, Larissa, V;Niemczura, Magdalena;van Dissel, Dino;van Wezel, Gilles P.;Thorson, Jon S.;Metsa-Ketela, Mikko;Shaaban, Khaled A.;Nybo, S. Eric
关键词：
放线菌产生多种临床上不可缺少的分子，例如抗肿瘤蒽环类药物。然而，由于与菌丝生命周期相关的缓慢生长动力学以及缺乏用于组合生物合成的综合遗传工具箱，放线菌作为合成新蒽环类药物类似物的基因工程宿主的进一步发展受到阻碍。在本报告中，我们通过开发 BIOPOLYMER（BIOBricks POLYketide Metabolic EngineeRing）工具箱解决了这两个问题：这是一个综合性合成生物学工具箱，由用于合成蒽环酮的工程菌株、启动子、载体和生物合成基因组成。通过删除指定细胞外聚-β-1,6-N-乙酰葡糖胺 (PNAG) 的 matAB 基因簇，创建了生产宿主天蓝色链霉菌 M1152 的改进衍生物。这导致菌丝聚集减少，生物量积累和蒽环酮产量提高。然后，我们利用 BIOPOLYMER 设计了四种不同的蒽环酮途径，确定了启动子、基因和载体的最佳组合，以生产滴度在 15–20 mg/L 之间的阿克拉维酮、9-表阿克拉维酮、金霉素酮和诺加霉素酮。诺加霉素生产菌株的优化导致效价为 103 mg/L。我们对发酵过程中的六种蒽环酮产品进行了结构表征，包括新化合物 9,10-seco-7-deoxy-nogalamycinone 和 4-O-β-d-glucosyl-nogalamycinone。最后，我们在哺乳动物癌细胞活力测定中测试了蒽环类药物的抗增殖活性，其中诺加霉素酮、金霉素酮和阿克拉维酮对几种癌细胞系表现出中等的细胞毒性。我们预计 BIOPOLYMER 将作为合成设计蒽环类类似物的基础平台技术。
Actinomycetes produce a variety of clinically indispensable molecules, such as antineoplastic anthracyclines. However, the actinomycetes are hindered in their further development as genetically engineered hosts for the synthesis of new anthracycline analogues due to their slow growth kinetics associated with their mycelial life cycle and the lack of a comprehensive genetic toolbox for combinatorial biosynthesis. In this report, we tackled both issues via the development of the BIOPOLYMER (BIOBricks POLYketide Metabolic EngineeRing) toolbox: a comprehensive synthetic biology toolbox consisting of engineered strains, promoters, vectors, and biosynthetic genes for the synthesis of anthracyclinones. An improved derivative of the production host Streptomyces coelicolor M1152 was created by deleting the matAB gene cluster that specifies extracellular poly-β-1,6-N-acetylglucosamine (PNAG). This resulted in a loss of mycelial aggregation, with improved biomass accumulation and anthracyclinone production. We then leveraged BIOPOLYMER to engineer four distinct anthracyclinone pathways, identifying optimal combinations of promoters, genes, and vectors to produce aklavinone, 9-epi-aklavinone, auramycinone, and nogalamycinone at titers between 15–20 mg/L. Optimization of nogalamycinone production strains resulted in titers of 103 mg/L. We structurally characterized six anthracyclinone products from fermentations, including new compounds 9,10-seco-7-deoxy-nogalamycinone and 4-O-β-d-glucosyl-nogalamycinone. Lastly, we tested the antiproliferative activity of the anthracyclinones in a mammalian cancer cell viability assay, in which nogalamycinone, auramycinone, and aklavinone exhibited moderate cytotoxicity against several cancer cell lines. We envision that BIOPOLYMER will serve as a foundational platform technology for the synthesis of designer anthracycline analogues.
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影响因子：
2.1
作者：
Flett, F;Mersinias, V;Smith, CP
通讯作者：
Smith, CP
DOI：
10.1073/pnas.1220220110
发表时间：
2013-05-07
影响因子：
11.1
作者：
Gantt, Richard W.;Peltier-Pain, Pauline;Thorson, Jon S.
通讯作者：
Thorson, Jon S.
影响因子：
3.3
作者：
Hautala, A;Torkkell, S;Ylihonko, K
通讯作者：
Ylihonko, K
影响因子：
3.5
作者：
BIBB, MJ;JANSSEN, GR;WARD, JM
通讯作者：
WARD, JM
影响因子：
2.9
作者：
FU, H;MCDANIEL, R;KHOSLA, C
通讯作者：
KHOSLA, C