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Heterologous Production of Polyunsaturated Fatty Acids in E. coli Using Δ5-Desaturase Gene from Microalga Isochrysis Sp.

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Abstract

Eicosapentaenoic acid (EPA) and arachidonic acid (ARA) are long-chain polyunsaturated fatty acids (PUFAs) that play a significant role in human growth and development, which deficiency can trigger several metabolic-related diseases. Since the availability of PUFA sources is limited, there arises a need to explore alternative sources. Therefore, the present study aimed to investigate whether an Escherichia coli which are engineered with Δ5Des-Iso gene isolated from Isochrysis sp. could be utilized to synthesize PUFAs. Full-length gene Δ5Des-Iso (1149 bp) was isolated from Isochrysis sp. that encodes 382 amino acids and identified as Δ5-desatruase gene using different bioinformatic analysis. Heterologous gene expression was carried out in E. coli having Δ5Des-Iso with precursor fatty acids. The Δ5Des-Iso produced novel fatty acids of EPA (ω-3) and ARA (ω-6) as respective products were identified by GC-MS. Gene expression and PUFA synthesis in E. coli were optimized by temperature, time, and concentrations of precursor fatty acid substrates. Δ5Des-Iso RNA transcript level was inversely proportional to the time and fatty acid synthesis. And, the significant production of EPA (4.1 mg/g) and ARA (8.3 mg/g) in total fatty acids was observed in E. coli grown at 37 °C for 24 h with 25 μM of external fatty acid substrate as an optimum growth conditions. E. coli could be used as alternative organism to synthesis PUFAs and widely applicable in many nutraceuticals and pharmaceuticals industry for human use.

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Acknowledgments

The authors would like to thank DBT-IPLS, NRCBS, CEGS, DST-PURSE, and UGC-CAS Phase III, School of Biological Sciences, Madurai Kamaraj University, Tamil Nadu, India, for the instrumentation facilities and the supports provided for this study. SK profoundly thanks Dr. Vikas Jain, Department of Biological Sciences, Indian Institute of Science Education and Research (IISER), Bhopal, India, for providing us with laboratory facilities to carry out Western blotting.

Funding

This work was supported by grants from Science and Engineering Research Board (SERB), Government of India (SB/EMEQ-219/2014, ECR/2017/002914).

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Authors and Affiliations

  1. Department of Molecular Biology, School of Biological Sciences, Madurai Kamaraj University, Madurai, Tamil Nadu, 625021, India

    S. Thiyagarajan, Priyal Khandelwal & S. Kathiresan

  2. Department of Plant Molecular Biology and Bioinformatics, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, 641003, India

    N. Senthil

  3. Department of Biotechnology, Centre of Innovation, Agriculture College and Research Institute, Madurai, Tamil Nadu, 625104, India

    S. Vellaikumar

  4. Microbial Processes and Technology Division, National Institute for Interdisciplinary Science and technology (NIIST), Council of Scientific and Industrial Research (CSIR), Trivandrum, Kerala, 695 019, India

    M. Arumugam

  5. Academy of Scientific and Innovative Research (AcSIR), New Delhi, 110001, India

    M. Arumugam

  6. Microbiology and Molecular Biology Laboratory, Department of Biological Sciences, Indian Institute of Science Education and Research (IISER), Bhopal, Madhya Pradesh, 462066, India

    Abhishek Anil Dubey

  7. Department of Life Sciences, School of Life Sciences, Central University of Tamil Nadu, Thiruvarur, Tamil Nadu, 610005, India

    S. Kathiresan

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  1. S. Thiyagarajan
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Correspondence to S. Kathiresan.

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Thiyagarajan, S., Khandelwal, P., Senthil, N. et al. Heterologous Production of Polyunsaturated Fatty Acids in E. coli Using Δ5-Desaturase Gene from Microalga Isochrysis Sp.. Appl Biochem Biotechnol 193, 869–883 (2021). https://doi.org/10.1007/s12010-020-03460-1

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