Abstract
5-Hydroxytryptophan (5-HTP) is the precursor of the neurotransmitter serotonin and has been used for the treatment of various diseases such as depression, insomnia, chronic headaches, and binge eating associated obesity. The production of 5-HTP had been achieved in our previous report, by the development of a recombinant strain containing two plasmids for biosynthesis of L-tryptophan (L-trp) and subsequent hydroxylation. In this study, the L-trp biosynthetic pathway was further integrated into the E. coli genome, and the promoter strength of 3-deoxy-7-phosphoheptulonate synthase, which catalyzes the first step of L-trp biosynthesis, was engineered to increase the production of L-trp. Hence, the 5-HTP production could be manipulated by the regulation of copy number of L-trp hydroxylation plasmid. Finally, the 5-HTP production was increased to 1.61 g/L in the shaking flasks, which was 24% improvement comparing to the original producing strain, while the content of residual L-trp was successfully reduced from 1.66 to 0.2 g/L, which is beneficial for the downstream separation and purification. Our work shall promote feasible progresses for the industrial production of 5-HTP.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Birdsall TC (1998) 5-Hydroxytryptophan: a clinically-effective serotonin precursor. Altern Med Rev 3(4):271–280
Carkaci-Salli N, Flanagan JM, Martz MK, Salli U, Walther DJ, Bader M, Vrana KE (2006) Functional domains of human tryptophan hydroxylase 2 (hTPH2). J Biol Chem 281(38):28105–28112. https://doi.org/10.1074/jbc.M602817200
Cui D, Deng A, Bai H, Yang Z, Liang Y, Liu Z, Qiu Q, Wang L, Liu S, Zhang Y, Shi Y, Qi J, Wen T (2019) Molecular basis for feedback inhibition of tyrosine-regulated 3-deoxy-d-arabino-heptulosonate-7-phosphate synthase from Escherichia coli. J Struct Biol 206(3):322–334. https://doi.org/10.1016/j.jsb.2019.04.001
Espah Borujeni A, Channarasappa AS, Salis HM (2014) Translation rate is controlled by coupled trade-offs between site accessibility, selective RNA unfolding and sliding at upstream standby sites. Nucleic Acids Res 42(4):2646–2659. https://doi.org/10.1093/nar/gkt1139
Fitzpatrick PF (2003) Mechanism of aromatic amino acid hydroxylation. Biochemistry 42(48):14083–14091. https://doi.org/10.1021/bi035656u
Hara R, Kino K (2013) Enhanced synthesis of 5-hydroxy-l-tryptophan through tetrahydropterin regeneration. AMB Express 3(1):70. https://doi.org/10.1186/2191-0855-3-70
Ikemoto K, Sugimoto T, Murata S, Tazawa M, Nomura T, Ichinose H, Nagatsu T (2002) (6R)-5,6,7,8-tetrahydro-L-monapterin from Escherichia coli, a novel natural unconjugated tetrahydropterin. Biol Chem 383(2):325–330. https://doi.org/10.1515/BC.2002.035
Jiang Y, Chen B, Duan C, Sun B, Yang J, Yang S (2015) Multigene editing in the Escherichia coli genome via the CRISPR-Cas9 system. Appl Environ Microb 81(7):2506–2514. https://doi.org/10.1128/AEM.04023-14
Keasling JD (2010) Manufacturing molecules through metabolic engineering. Science 330(6009):1355–1358. https://doi.org/10.1126/science.1193990
Kino K, Hara R, Nozawa A (2009) Enhancement of L-tryptophan 5-hydroxylation activity by structure-based modification of L-phenylalanine 4-hydroxylase from Chromobacterium violaceum. J Biosci Bioeng 108(3):184–189. https://doi.org/10.1016/j.jbiosc.2009.04.002
Knight EM, Zhu J, Förster J, Luo H (2013) Microorganisms for the production of 5-hydroxytryptophan. WO2013127914A1
Lim HG, Noh MH, Jeong JH, Park S, Jung GY (2016) Optimum rebalancing of the 3-hydroxypropionic acid production pathway from glycerol in Escherichia coli. ACS Synth Biol 5(11):1247–1255. https://doi.org/10.1021/acssynbio.5b00303
Lin Y, Sun X, Yuan Q, Yan Y (2014) Engineering bacterial phenylalanine 4-hydroxylase for microbial synthesis of human neurotransmitter precursor 5-hydroxytryptophan. ACS Synth Biol 3(7):497–505. https://doi.org/10.1021/sb5002505
Lu J, Tang J, Liu Y, Zhu X, Zhang T, Zhang X (2012) Combinatorial modulation of galP and glk gene expression for improved alternative glucose utilization. Appl Microbiol Biotechnol 93(6):2455–2462. https://doi.org/10.1007/s00253-011-3752-y
Mora-Villalobos JA, Zeng AP (2018) Synthetic pathways and processes for effective production of 5-hydroxytryptophan and serotonin from glucose in Escherichia coli. J Biol Eng 12:3. https://doi.org/10.1186/s13036-018-0094-7
Mora-Villalobos JA, Zeng AP (2017) Protein and pathway engineering for the biosynthesis of 5-hydroxytryptophan in Escherichia coli. Eng Life Sci 17:892–899. https://doi.org/10.1002/elsc.201700064
Murphy KL, Zhang X, Gainetdinov RR, Beaulieu JM, Caron MG (2008) A regulatory domain in the N terminus of tryptophan hydroxylase 2 controls enzyme expression. J Biol Chem 283(19):13216–13224. https://doi.org/10.1074/jbc.M706749200
Nielsen MS, Petersen CR, Munch A, Vendelboe TV, Boesen J, Harris P, Christensen HE (2008) A simple two step procedure for purification of the catalytic domain of chicken tryptophan hydroxylase 1 in a form suitable for crystallization. Protein Expr Purif 57(2):116–126. https://doi.org/10.1016/j.pep.2007.10.016
World Health Organization (2017) Depression and other common mental disorders: global health estimates. https://apps.who.int/iris/handle/10665/254610
Pribat A, Blaby IK, Lara-Nunez A, Gregory JF, Crecy-Lagard V, Hanson AD (2010) FolX and FolM are essential for tetrahydromonapterin synthesis in Escherichia coli and Pseudomonas aeruginosa. J Bacteriol 192(2):475–482. https://doi.org/10.1128/JB.01198-09
Ran N, Frost JW (2007) Directed evolution of 2-keto-3-deoxy-6-phosphogalactonate aldolase to replace 3-deoxy-d-arabino-heptulosonic acid 7-phosphate synthase. J Am Chem Soc 129(19):6130–6139. https://doi.org/10.1021/ja067330p
Turner EH, Loftis JM, Blackwell AD (2006) Serotonin a la carte: supplementation with the serotonin precursor 5-hydroxytryptophan. Pharmacol Ther 109(3):325–338. https://doi.org/10.1016/j.pharmthera.2005.06.004
Wang H, Liu W, Shi F, Huang L, Lian J, Qu L, Cai J, Xu Z (2018) Metabolic pathway engineering for high-level production of 5-hydroxytryptophan in Escherichia coli. Metab Eng 48:279–287. https://doi.org/10.1016/j.ymben.2018.06.007
Windahl MS, Boesen J, Karlsen PE, Christensen HE (2009) Expression, purification and enzymatic characterization of the catalytic domains of human tryptophan hydroxylase isoforms. Protein J 28(9–10):400–406. https://doi.org/10.1007/s10930-009-9207-y
Xu P, Gu Q, Wang W, Wong L, Bower A, Collins CH, Ma K (2013) Modular optimization of multi-gene pathways for fatty acids production in E. coli. Nat Commun 4(1):1409. https://doi.org/10.1038/ncomms2425
Funding
This work was financially supported by the National Natural Science Foundation of China (Nos. 21576232, 21606205, and 21808199), the National Key Research and Development Program of China (No. 2018YFC1603900), and the Natural Science Foundation of Zhejiang Province (No. LY18B060002).
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflict of interest
All authors confirm that they have no conflict of interests.
Human and animal rights
This article does not contain any studies with human participants or animals performed by any of the authors.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
ESM 1
(PDF 371 kb)
Rights and permissions
About this article
Cite this article
Xu, D., Fang, M., Wang, H. et al. Enhanced production of 5-hydroxytryptophan through the regulation of L-tryptophan biosynthetic pathway. Appl Microbiol Biotechnol 104, 2481–2488 (2020). https://doi.org/10.1007/s00253-020-10371-y
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-020-10371-y