Abstract
TetR family transcriptional regulators (TFRs) are widespread in actinomycetes, which exhibit diverse regulatory modes in antibiotic biosynthesis. Nitrogen regulators play vital roles in modulation of primary and secondary metabolism. However, crosstalk between TFR and nitrogen regulator has rarely been reported in actinomycetes. Herein, we demonstrated that a novel TFR, SACE_4839, was negatively correlated with erythromycin yield in Saccharopolyspora erythraea A226. SACE_4839 indirectly suppressed erythromycin synthetic gene eryAI and resistance gene ermE and directly inhibited its adjacent gene SACE_4838 encoding a homologue of nitrogen metabolite repression (NMR) regulator NmrA (herein named NmrR). The SACE_4839-binding sites within SACE_4839-nmrR intergenic region were identified. NmrR positively controlled erythromycin biosynthesis by indirectly stimulating eryAI and ermE and directly repressing SACE_4839. NmrR was found to affect growth viability under the nitrogen source supply. Furthermore, NmrR directly repressed glutamine and glutamate utilization-related genes SACE_1623, SACE_5070 and SACE_5979 but activated nitrate utilization-associated genes SACE_1163, SACE_4070 and SACE_4912 as well as nitrite utilization-associated genes SACE_1476 and SACE_4514. This is the first reported NmrA homolog for modulating antibiotic biosynthesis and nitrogen metabolism in actinomycetes. Moreover, combinatorial engineering of SACE_4839 and nmrR in the high-yield S. erythraea WB resulted in a 68.8% increase in erythromycin A production. This investigation deepens the understanding of complicated regulatory network for erythromycin biosynthesis.
Key points
• SACE_4839 and NmrR had opposite contributions to erythromycin biosynthesis.
• NmrR was first identified as a homolog of another nitrogen regulator NmrA.
• Cross regulation between SACE_4839 and NmrR was revealed.
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All the data generated or analyzed during this study are included in this published article (and its supplementary information files).
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Funding
This study was supported by the National Natural Science Foundation of China (grant numbers 31972930, 32170073 and 31800057), the Anhui Provincial Natural Science Foundation (grant number 2208085Y09), the University Synergy Innovation Program of Anhui Province (grant number GXXT-2019–035), and the Anhui Provincial Program on Key Research and Development Project (grant number 201904a07020080).
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BZ, HW and PW conceived and designed the study. SK, XX, JS, XL, KC and ZX performed the experiments. SK, XX, JS, PW, JL and HW analyzed the data. SK and PW wrote the manuscript. HW and BZ checked the final version. All the authors read and approved the final manuscript.
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Khan, S., Xu, X., Song, J. et al. Crosstalk of TetR-like regulator SACE_4839 and a nitrogen regulator for erythromycin biosynthesis. Appl Microbiol Biotechnol 106, 6551–6566 (2022). https://doi.org/10.1007/s00253-022-12153-0
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DOI: https://doi.org/10.1007/s00253-022-12153-0