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Engineering of an Lrp family regulator SACE_Lrp improves erythromycin production in Saccharopolyspora erythraea.
- Source :
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Metabolic engineering [Metab Eng] 2017 Jan; Vol. 39, pp. 29-37. Date of Electronic Publication: 2016 Oct 26. - Publication Year :
- 2017
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Abstract
- 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&#95;5388, an Lrp family regulator named as SACE&#95;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&#95;Lrp directly controlled the expression of the divergently transcribed SACE&#95;5387-5386 operon putatively encoding a BCAA ABC transporter by interacting with the intergenic region between SACE&#95;Lrp and SACE&#95;5387 (SACE&#95;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&#95;Lrp from SACE&#95;Lrp -5387-int, whereas histidine increased their binding. Gene disruption of SACE&#95;Lrp (ΔSACE&#95;Lrp) in S. erythraea A226 resulted in a 25% increase in erythromycin production, while overexpression of SACE&#95;5387-5386 in A226 enhanced erythromycin production by 36%. Deletion of SACE&#95;Lrp (WBΔSACE&#95;Lrp) in the industrial strain S. erythraea WB enhanced erythromycin production by 19%, and overexpression of SACE&#95;5387-5386 in WBΔSACE&#95;Lrp (WBΔSACE&#95;Lrp/5387-5386) increased erythromycin production by 41% compared to WB. Additionally, supplement of 10mM valine to WBΔSACE&#95;Lrp/5387-5386 culture further increased total erythromycin production up to 48%. In a 5-L fermenter, the erythromycin accumulation in the engineered strain WBΔSACE&#95;Lrp/5387-5386 with 10mM extra valine in the industrial culture media reached 5001mg/L, a 41% increase over 3503mg/L of WB. These insights into the molecular regulation of antibiotic biosynthesis by SACE&#95;Lrp in S. erythraea are instrumental in increasing industrial production of secondary metabolites.<br /> (Copyright © 2016 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.)
- Subjects :
- Bacterial Proteins genetics
Erythromycin isolation & purification
Metabolic Networks and Pathways genetics
Biosynthetic Pathways genetics
Erythromycin biosynthesis
Genetic Enhancement methods
Leucine-Responsive Regulatory Protein genetics
Metabolic Engineering methods
Saccharopolyspora physiology
Subjects
Details
- Language :
- English
- ISSN :
- 1096-7184
- Volume :
- 39
- Database :
- MEDLINE
- Journal :
- Metabolic engineering
- Publication Type :
- Academic Journal
- Accession number :
- 27794466
- Full Text :
- https://doi.org/10.1016/j.ymben.2016.10.012