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Production of serotonin by dual expression of tryptophan decarboxylase and tryptamine 5-hydroxylase in Escherichia coli

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Abstract

A plant-specific biogenic amine, serotonin, was produced by heterologous expression of two key biosynthetic genes, tryptophan decarboxylase (TDC) and tryptamine 5-hydroxylase (T5H), in Escherichia coli. The native T5H, a cytochrome P450 enzyme, was unable to be functionally expressed in E. coli. Through a series of N-terminal deletions or additions of tagging proteins, we generated a functional T5H enzyme construct (GST∆37T5H) in which glutathione S transferase (GST) was translationally fused with the N-terminal 37 amino acid deleted T5H. Dual expression of GST∆37T5H and TDC using a pCOLADuet-1 E. coli vector produced serotonin at concentrations of approximately 24 mg l−1 in the culture medium and 4 mg l−1 in the cells. An optimum temperature of approximately 20°C was required to achieve peak serotonin production in E. coli because the low induction temperature gave rise to the highest soluble expression of GST∆37T5H.

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Acknowledgments

This work was supported by the Technology Development Program for Agriculture and Forestry, Ministry of Agriculture and Forestry (108078-3), the BioGreen 21 Program (20100301-061-023-001-03-00) funded by the Rural Development Administration, and the Priority Research Centers Program through the National Research Foundation (NRF) funded by the Ministry of Education, Science, and Technology (2010-0020141).

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Authors and Affiliations

  1. Department of Biotechnology, Interdisciplinary Programs of Graduate School for Bioenergy and Biomaterials, Bioenergy Research Center, Chonnam National University, Gwangju, 500-757, South Korea

    Sangkyu Park, Kiyoon Kang & Kyoungwhan Back

  2. Department of Crop Science & Biotechnology, Jinju National University, Jinju, 660-758, South Korea

    Shin Woo Lee & Mi-Jeong Ahn

  3. School of Life Sciences and Biotechnology, Korea University, Seoul, 136-701, South Korea

    Jung-Myung Bae

  4. Department of Biotechnology, College of Agriculture and Life Sciences, Chonnam National University, Gwangju, 500-757, Republic of Korea

    Kyoungwhan Back

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  1. Sangkyu Park
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  2. Kiyoon Kang
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  3. Shin Woo Lee
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  4. Mi-Jeong Ahn
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Correspondence to Kyoungwhan Back.

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Park, S., Kang, K., Lee, S.W. et al. Production of serotonin by dual expression of tryptophan decarboxylase and tryptamine 5-hydroxylase in Escherichia coli . Appl Microbiol Biotechnol 89, 1387–1394 (2011). https://doi.org/10.1007/s00253-010-2994-4

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