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Expression of soluble moloney murine leukemia virus-reverse transcriptase in Escherichia coli BL21 star (DE3) using autoinduction system

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Abstract

Autoinduction systems in Escherichia coli can control the production of proteins without the addition of a particular inducer. In the present study, we optimized the heterologous expression of Moloney Murine Leukemia Virus derived Reverse Transcriptase (MMLV-RT) in E. coli. Among 4 autoinduction media, media Imperial College resulted the highest MMLV-RT overexpression in E. coli BL21 Star (DE3) with incubation time 96 h. The enzyme was produced most optimum in soluble fraction of lysate cells. The MMLV-RT was then purified using the Immobilized Metal Affinity Chromatography method and had specific activity of 629.4 U/mg. The system resulted lower specific activity and longer incubation of the enzyme than a classical Isopropyl ß-D-1-thiogalactopyranoside (IPTG)-induction system. However, the autoinduction resulted higher yield of the enzyme than the conventional induction (27.8%). Techno Economic Analysis revealed that this method could produce MMLV-RT using autoinduction at half the cost of MMLV-RT production by IPTG-induction. Bioprocessing techniques are necessary to conduct to obtain higher quality of MMLV-RT under autoinduction system.

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Acknowledgements

The authors would like to thank the Research Team of Extremophile Enzyme, Research Center of Applied Microbiology, National Research and Innovation Agency for permission, guidance, and facilities.

Funding

This research was supported by Indonesia Endowment Fund for Education (LPDP) Program in fiscal year 2021 for National Priority Program for COVID-19.

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Authors and Affiliations

  1. Research Center for Applied Microbiology, National Agency for Research and Innovation, Jl. Raya Bogor, Km. 46, Cibinong, Bogor, 16911, Indonesia

    Christina Vivid Handayani, Fina Amreta Laksmi, Ade Andriani, Isa Nuryana, Eva Agustriana & Awan Purnawan

  2. Biotechnology Program, Graduate School, IPB University, Bogor, Indonesia

    Christina Vivid Handayani

  3. Department of Biology, Faculty of Mathematic and Natural Science, IPB University, Bogor, Indonesia

    Nisa Rachmania Mubarik

  4. Research Center for Genetic Engineering, National Agency for Research and Innovation, Jl. Raya Bogor, Km. 46, Cibinong, Bogor, 16911, Indonesia

    Kartika Sari Dewi

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  1. Christina Vivid Handayani
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  2. Fina Amreta Laksmi
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  3. Ade Andriani
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Contributions

CV carried out the research, analyzed and interpreted the data, and wrote the manuscript, FAL designed the research and reviewed the manuscript, IN analyzed and interpreted the data and wrote the manuscript, AA provided supervision, wrote and reviewed the manuscript, NR provided supervision and reviewed the manuscript, EA wrote the manuscript, AP reviewed the manuscript.

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Correspondence to Fina Amreta Laksmi or Ade Andriani.

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Handayani, C.V., Laksmi, F.A., Andriani, A. et al. Expression of soluble moloney murine leukemia virus-reverse transcriptase in Escherichia coli BL21 star (DE3) using autoinduction system. Mol Biol Rep 51, 628 (2024). https://doi.org/10.1007/s11033-024-09583-6

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