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4,4′-Diaponeurosporene Production as C30 Carotenoid with Antioxidant Activity in Recombinant Escherichia coli

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Abstract

Carotenoids, a group of isoprenoid pigments, are naturally synthesized by various microorganisms and plants, and are industrially used as ingredients in food, cosmetic, and pharmaceutical product formulations. Although several types of carotenoids and diverse microbial carotenoid producers have been reported, studies on lactic acid bacteria (LAB)-derived carotenoids are relatively insufficient. There is a notable lack of research focusing on C30 carotenoids, the functional characterizations of their biosynthetic genes and their mass production by genetically engineered microorganisms. In this study, the biosynthesis of 4,4′-diaponeurosporene in Escherichia coli harboring the core biosynthetic genes, dehydrosqualene synthase (crtM) and dehydrosqualene desaturase (crtN), from Lactiplantibacillus plantarum subsp. plantarum KCCP11226 was constructed to evaluate and enhance 4,4′-diaponeurosporene production and antioxidant activity. The production of 4,4′-diapophytoene, a substrate of 4,4′-diaponeurosporene, was confirmed in E. coli expressing only the crtM gene. In addition, recombinant E. coli carrying both C30 carotenoid biosynthesis genes (crtM and crtN) was confirmed to biosynthesize 4,4′-diaponeurosporene and exhibited a 6.1-fold increase in carotenoid production compared to the wild type and had a significantly higher antioxidant activity compared to synthetic antioxidant, butylated hydroxytoluene. This study presents the discovery of an important novel E. coli platform in consideration of the industrial applicability of carotenoids.

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Funding

This work was supported by the National Research Foundation of Korea (NRF) grants funded by the Korea government (MIST) (Grant No. 2019R1A2C1006038 and 2022R1F1A1062699). This research was also supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (Grant No. 2021R1A6A3A01087585). In addition, this research was supported by the Ottogi Ham Taiho Foundation.

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Authors and Affiliations

  1. Department of Chemical Engineering, Pohang University of Science and Technology, Pohang, 37673, Gyeongbuk, Korea

    Mibang Kim

  2. Department of Bioengineering and Nano-Bioengineering, Incheon National University, Incheon, 22012, Republic of Korea

    Mibang Kim, Chi Young Hwang & Myung-Ji Seo

  3. Microorganism Resources Division, National Institute of Biological Resources, Incheon, 22689, Republic of Korea

    Dong-Hyun Jung

  4. Division of Bioengineering, Incheon National University, Incheon, 22012, Republic of Korea

    Inonge Noni Siziya & Myung-Ji Seo

  5. Research Center for Bio Material & Process Development, Incheon National University, Incheon, 22012, Republic of Korea

    Inonge Noni Siziya & Myung-Ji Seo

  6. Department of Food Science and Biotechnology, Gachon University, Seongnam, 13120, Republic of Korea

    Young-Seo Park

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  1. Mibang Kim
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Contributions

All authors contributed to the study conception and design. Myung-Ji Seo designed and coordinated all the experiments; Mibang Kim performed in vitro experiments on 4,4′-diaponeurosporene production in E. coli and wrote the manuscript draft; Dong-Hyun Jung and Young-Seo Park analyzed the research data and reviewed the manuscript. Chi Young Hwang and Inonge Noni Siziya performed antioxidant activity of 4,4′-diaponeurosporene and analyzed the carotenoid production level. All authors have read and approved the final manuscript.

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Correspondence to Myung-Ji Seo.

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Kim, M., Jung, DH., Hwang, C.Y. et al. 4,4′-Diaponeurosporene Production as C30 Carotenoid with Antioxidant Activity in Recombinant Escherichia coli. Appl Biochem Biotechnol 195, 135–151 (2023). https://doi.org/10.1007/s12010-022-04147-5

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