Joint engineering of SACE_Lrp and its target MarR enhances the biosynthesis and export of erythromycin in Saccharopolyspora erythraea
Joint engineering of SACE_Lrp and its target MarR enhances the biosynthesis and export of erythromycin in Saccharopolyspora erythraea
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SACE_Lrp 及其靶点 MarR 的联合工程增强了红糖多孢菌中红霉素的生物合成和输出
DOI：
10.1007/s00253-021-11228-8
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发表时间：
2021-03-24
影响因子：
5
通讯作者：
Zhang, Buchang
中科院分区：
文献类型：
--
作者：
Liu, Jing;Li, Long;Zhang, Buchang
Lrp和MarR家族是广泛分布于原核生物中的两组转录调控因子。然而，Lrp家族和MarR家族之间的等级调节关系尚不清楚。我们前期的研究发现，来自红糖多孢子菌的Lrp （SACE_Lrp）间接抑制红霉素的生物合成。在本研究中，我们从S. erythraea中鉴定了一个新的MarR家族蛋白（SACE_6745），该蛋白受SACE_Lrp控制，在红霉素的生物合成和输出中起直接调节作用。SACE_Lrp通过特异性结合精确位点OM（5’- ctccgggaaccat -3’）直接调控marR的表达。marR基因的破坏使红链球菌A226的红霉素产量增加了45%。我们发现MarR对红霉素生物合成基因的启动子区域具有直接的dna结合活性，以及一个ABC出口者SACE_2701-2702，该基因被遗传学证明是红霉素外流的原因。破坏工业红脓杆菌的SACE_Lrp是提高红霉素产量的有效策略。本文中，我们通过删除WB Delta SACE_Lrp中的MarR，对SACE_Lrp及其靶MarR进行了联合改造，结果突变体WB Delta Lrp Delta MarR的红霉素产量比WB Delta SACE_Lrp增加了20%，比WB Delta SACE_Lrp增加了39%。总的来说，我们的研究结果为Lrp和MarR蛋白的层次调控关系提供了新的见解，并为放线菌中联合工程调控协调抗生素的生物合成和出口提供了新的途径。
The Lrp and MarR families are two groups of transcriptional regulators widely distributed among prokaryotes. However, the hierarchical-regulatory relationship between the Lrp family and the MarR family remains unknown. Our previous study found that an Lrp (SACE_Lrp) from Saccharopolyspora erythraea indirectly repressed the biosynthesis of erythromycin. In this study, we characterized a novel MarR family protein (SACE_6745) from S. erythraea, which is controlled by SACE_Lrp and plays a direct regulatory role in erythromycin biosynthesis and export. SACE_Lrp directly regulated the expression of marR by specifically binding a precise site OM (5'-CTCCGGGAACCATT-3'). Gene disruption of marR increased the production of erythromycin by 45% in S. erythraea A226. We found that MarR has direct DNA-binding activity for the promoter regions of the erythromycin biosynthetic genes, as well as an ABC exporter SACE_2701-2702 which was genetically proved to be responsible for erythromycin efflux. Disruption of SACE_Lrp in industrial S. erythraea WB was an efficient strategy to enhance erythromycin production. Herein, we jointly engineered SACE_Lrp and its target MarR by deleting marR in WB Delta SACE_Lrp, resulting in 20% increase in erythromycin yield in mutant WB Delta Lrp Delta marR compared to WB Delta SACE_Lrp, and 39% to WB. Overall, our findings provide new insights into the hierarchical-regulatory relationship of Lrp and MarR proteins and new avenues for coordinating antibiotic biosynthesis and export by joint engineering regulators in actinomycetes.