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One-pot synthesis of dihydro-β-ionone from carotenoids using carotenoid cleavage dioxygenase and enoate reductase

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Abstract

Dihydro-β-ionone is a characteristic aroma compound of Osmanthus fragrans and is widely applied in the flavor & fragrance industry. However, the main focus is on chemical synthesis due to the metabolic pathways of dihydro-β-ionone is still unclear. Here, we explored the one-pot synthesis system for dihydro-β-ionone production using carotenoid cleavage dioxygenase (CCD) and enoate reductase. After screening the CCD enzyme, PhCCD1 from the Petunia hybrid was identified as the suitable enzyme for the first step of dihydro-β-ionone synthesis due to the high enzyme activity for carotenoid. The PhCCD1 was expressed in Escherichia coli and further characterized. The optimal activity of PhCCD1 was observed at pH 6.8 and 45 °C. The enzyme was stable over the pH range of 6.0–8.0 and had good thermal stability below 40 °C. Then, we optimized the coupled reaction conditions for dihydro-β-ionone production by PhCCD1 and enoate reductase AaDBR1 from Artemisia annua. Furthermore, we introduced the NADPH regeneration system with a 1.5-fold enhancement for dihydro-β-ionone production. Collectively, approximately 13.34 mg/L dihydro-β-ionone was obtained by the one-pot biosystem with a corresponding molar conversion of 85.8%. For the first time, we successfully designed and constructed a new synthesis pathway for dihydro-β-ionone production in vitro. The coupled catalysis reported herein illustrates the feasibility of producing dihydro-β-ionone from carotenoids and guides further engineering in the food industry.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant No. 32071711).

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  1. Zhipeng Qi and Xinyi Tong contributed equally to this work.

Authors and Affiliations

  1. Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, Nanjing Forestry University, 159 Long Pan Road, Nanjing, 210037, People’s Republic of China

    Zhipeng Qi, Xiaomeng Zhang & Linguo Zhao

  2. College of Chemical Engineering, Nanjing Forestry University, 159 Long Pan Road, Nanjing, 210037, People’s Republic of China

    Zhipeng Qi, Xinyi Tong, Xiaomeng Zhang, Haifeng Lin & Linguo Zhao

  3. College of Biology and the Environment, Nanjing Forestry University, Nanjing, 210037, Jiangsu, People’s Republic of China

    Su Bu

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  1. Zhipeng Qi
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Qi, Z., Tong, X., Zhang, X. et al. One-pot synthesis of dihydro-β-ionone from carotenoids using carotenoid cleavage dioxygenase and enoate reductase. Bioprocess Biosyst Eng 45, 891–900 (2022). https://doi.org/10.1007/s00449-022-02707-x

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