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Downregulating of hemB via synthetic antisense RNAs for improving 5-aminolevulinic acid production in Escherichia coli

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Abstract

Aminolevulinic acid (ALA), a type of natural non-protein amino acid, is a key precursor for the biosynthesis of heme, and it has been broadly applied in medicine, agriculture. Several strategies have been applied to enhance ALA synthesis in bacteria. In the present study, we employed synthetic antisense RNAs (asRNAs) of hemB (encodes ALA dehydratase) to weaken metabolic flux of ALA to porphobilinogen (PBG), and investigated their effect on ALA accumulation. For this purpose, we designed and constructed vectors pET28a-hemA-asRNA and pRSFDuet-hemA-asRNA to simultaneously express 5-ALA synthase (ALAS, encoded by hemA) and PTasRNAs (2 inverted repeat DNA sequences sandwiched with the antisense sequence of hemB), selecting the region ranging from − 57 nt upstream to + 139 nt downstream of the start codon of hemB as a target. The qRT-PCR analysis showed that the mRNA levels of hemB were decreased above 50% of the control levels, suggesting that the anti-hemB asRNA was functioning appropriately. ALA accumulation in the hemB weakened strains were 17.6% higher than that obtained using the control strains while accumulating less PBG. These results indicated that asRNAs can be used as a tool for regulating ALA accumulation in E. coli.

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Acknowledgements

This study was funded by the Sichuan provincial Science & Technology Department (2018JY0106, 2018JY0310).

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  1. Fanglan Ge and Dongmei Wen are equally contributed to this paper.

Authors and Affiliations

  1. College of Life Sciences, Sichuan Normal University, Chengdu, 610068, People’s Republic of China

    Fanglan Ge, Dongmei Wen, Yao Ren, Guiying Chen, Bing He, Xiaokun Li & Wei Li

  2. Key Laboratory for Utilization and Conservation of Bio-Resources of Education, Department of Sichuan Province, Chengdu, People’s Republic of China

    Wei Li

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  1. Fanglan Ge
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  2. Dongmei Wen
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  3. Yao Ren
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  4. Guiying Chen
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  6. Xiaokun Li
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  7. Wei Li
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Correspondence to Wei Li.

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None of the authors has any financial or personal relationships that could inappropriately influence or bias the content of the research paper. The authors declare that they have no conflict of interest in the publication.

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Ge, F., Wen, D., Ren, Y. et al. Downregulating of hemB via synthetic antisense RNAs for improving 5-aminolevulinic acid production in Escherichia coli. 3 Biotech 11, 230 (2021). https://doi.org/10.1007/s13205-021-02733-8

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  • DOI: https://doi.org/10.1007/s13205-021-02733-8

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