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Promoter Screening Facilitates Heterologous Production of Complex Secondary Metabolites in Burkholderiales Strains.
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2020-02-07 , DOI: 10.1021/acssynbio.9b00459 Qing Ouyang 1 , Xue Wang 1 , Na Zhang 1 , Lin Zhong 1 , Jiaqi Liu 1 , Xiaoming Ding 2 , Youming Zhang 1 , Xiaoying Bian 1
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2020-02-07 , DOI: 10.1021/acssynbio.9b00459 Qing Ouyang 1 , Xue Wang 1 , Na Zhang 1 , Lin Zhong 1 , Jiaqi Liu 1 , Xiaoming Ding 2 , Youming Zhang 1 , Xiaoying Bian 1
Affiliation
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Burkholderiales are an emerging source of bioactive secondary metabolites and have the potential to be a robust chassis for metabolites from Gram-negative bacteria. However, only a few constitutive promoters can be utilized in Burkholderiales. Herein, we described the screening of strong constitutive promoters from Burkholderiales strain DSM 7029, and 37 promoters identified from transcriptome sequencing were cloned and characterized using a firefly luciferase reporter and were further verified by qPCR analysis. These promoters were then used to drive a complex 56-kb epothilone BGC from myxobacterium and a 23-kb rhizomide BGC from Paraburkholderia rhizoxinica, and the successful production of epothilone and rhizomide was observed in DSM 7029, with improved yields compared to the production achieved by previously used promoters. Additionally, these promoters are also functional in other Burkholderiales species. Thus, these promoters are highly useful for optimizing yields of important metabolites in Burkholderiales and for mining cryptic biosynthetic pathways in DSM 7029.
中文翻译：
启动子筛选有助于在伯克霍尔德里亚氏菌菌株中异源产生复杂的次生代谢产物。
伯克霍尔德氏菌是生物活性次生代谢物的新兴来源，并有可能成为革兰氏阴性细菌代谢物的强大底盘。但是，伯克霍尔德里亚氏菌只能使用少数组成型启动子。在本文中，我们描述了从Burkholderiales菌株DSM 7029筛选强组成型启动子的过程，并使用萤火虫荧光素酶报告基因克隆了从转录组测序中鉴定的37个启动子，并通过qPCR分析进行了进一步验证。然后将这些启动子用于驱动来自粘杆菌的复杂的56kb埃博霉素BGC和来自根瘤菌的23kb根茎BGC，在DSM 7029中观察到埃坡霉素和根茎的成功生产，与通过以前使用的启动子。另外，这些启动子在其他伯克霍尔德里亚种中也有功能。因此，这些启动子对于优化伯克霍尔德氏菌中重要代谢物的产量以及在DSM 7029中挖掘隐秘生物合成途径非常有用。
更新日期：2020-02-10
中文翻译：
启动子筛选有助于在伯克霍尔德里亚氏菌菌株中异源产生复杂的次生代谢产物。
伯克霍尔德氏菌是生物活性次生代谢物的新兴来源，并有可能成为革兰氏阴性细菌代谢物的强大底盘。但是，伯克霍尔德里亚氏菌只能使用少数组成型启动子。在本文中，我们描述了从Burkholderiales菌株DSM 7029筛选强组成型启动子的过程，并使用萤火虫荧光素酶报告基因克隆了从转录组测序中鉴定的37个启动子，并通过qPCR分析进行了进一步验证。然后将这些启动子用于驱动来自粘杆菌的复杂的56kb埃博霉素BGC和来自根瘤菌的23kb根茎BGC，在DSM 7029中观察到埃坡霉素和根茎的成功生产，与通过以前使用的启动子。另外，这些启动子在其他伯克霍尔德里亚种中也有功能。因此，这些启动子对于优化伯克霍尔德氏菌中重要代谢物的产量以及在DSM 7029中挖掘隐秘生物合成途径非常有用。
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