Engineering of an Lrp family regulator SACE_Lrp improves erythromycin production in Saccharopolyspora erythraea
Engineering of an Lrp family regulator SACE_Lrp improves erythromycin production in Saccharopolyspora erythraea
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Lrp 家族调节剂 SACE_Lrp 的工程设计提高了红糖多孢菌中红霉素的产量
DOI：
10.1016/j.ymben.2016.10.012
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发表时间：
2017-01-01
影响因子：
8.4
通讯作者：
Zhang, Buchang
中科院分区：
文献类型：
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作者：
Liu, Jing;Chen, Yunfu;Zhang, Buchang
亮氨酸应答调节蛋白（Leucine-responsive regulatory proteins，Lrps）是一类在细菌和古细菌中调节多种细胞过程的转录调节因子。然而，Lrps在抗生素生物合成中的调节作用仍然知之甚少。在这项研究中，我们发现，SACE_5388，一个Lrp家族的调节剂命名为SACE_Lrp，是一个有效的调节剂，运输和分解代谢支链氨基酸（BCAAs），发挥重要作用，在红霉素生产中起着重要的作用，在多孢霉。SACE_Lrp通过与SACS_Lrp和SAGE 5387之间的基因间隔区（SACE_Lrp-5387-int）相互作用，直接控制编码BCAA ABC转运蛋白的差异转录的SACE_5387-5386操纵子pupp 3的表达，并间接控制编码BCAA的氨基转移酶分解代谢的ilvE pupp 3的表达。支链氨基酸催化剂是红霉素生物合成的前体物质之一。赖氨酸和精氨酸促进SACE_Lrp与SACE_Lrp-5387-int的解离，而组氨酸增加它们的结合。S. SACE_Lrp（Delta SACE_Lrp）的基因中断。拟南芥A226导致红霉素产量增加25%，而SACS 5387-5386在A226中的过表达使红霉素产量增加36%。在工业菌株S中缺失SACE_ Lrp（WB Delta SACE_Lrp）。与WB相比，拟南芥WB使红霉素产量增加19%，并且WB Delta SACE_Lrp中SACE_ 5387-5386的过表达（WB Delta SACE_Lrp/5387-5386）使红霉素产量增加41%。此外，向WB Delta SACE_Lrp/5387-5386培养物中补充10 mM缬氨酸进一步将总红霉素产量增加至48%。在5L发酵罐中，工程菌株WB Delta SACE_Lrp/5387-5386在工业培养基中添加10 mM额外缬氨酸时，红霉素积累达到5001 mg/L，比WB的3503 mg/L增加41%。SACE_Lrp对S.在增加次级代谢产物的工业生产中，藜麦是有用的。
Leucine-responsive regulatory proteins (Lrps) are a group of transcriptional regulators that regulate diverse cellular processes in bacteria and archaea. However, the regulatory role of Lrps in antibiotic biosynthesis remains poorly understood. In this study, we show that SACE_5388, an Lrp family regulator named as SACE_Lrp, is an efficient regulator for transporting and catabolizing branched-chain amino acids (BCAAs), playing an important role in regulating erythromycin production in Saccharopolyspora erythraea. SACE_Lrp directly controlled the expression of the divergently transcribed SACE_5387-5386 operon putatively encoding a BCAA ABC transporter by interacting with the intergenic region between SACS_Lrp and SAGE 5387 (SACE_Lrp-5387-int), and indirectly controlled the expression of ilvE putatively encoding an aminotransferase catabolizing BCAAs. BCAA catabolism is one source of the precursors for erythromycin biosynthesis. Lysine and arginine promoted the dissociation of SACE_Lrp from SACE_Lrp -5387-int, whereas histidine increased their binding. Gene disruption of SACE_Lrp (Delta SACE_Lrp) in S. erythraea A226 resulted in a 25% increase in erythromycin production, while overexpression of SACS 5387-5386 in A226 enhanced erythromycin production by 36%. Deletion of SACE_ Lrp (WB Delta SACE_Lrp) in the industrial strain S. erythraea WB enhanced erythromycin production by 19%, and overexpression of SACE_ 5387-5386 in WB Delta SACE_Lrp (WB Delta SACE_Lrp/5387-5386) increased erythromycin production by 41% compared to WB. Additionally, supplement of 10mM valine to WB Delta SACE_Lrp/5387-5386 culture further increased total erythromycin production up to 48%. In a 5-L fermenter, the erythromycin accumulation in the engineered strain WB Delta SACE_Lrp/5387-5386 with 10 mM extra valine in the industrial culture media reached 5001 mg/L, a 41% increase over 3503 mg/L of WB. These insights into the molecular regulation of antibiotic biosynthesis by SACE_Lrp in S. erythraea are instrumental in increasing industrial production of secondary metabolites.