Heterogeneous expression of DOPA decarboxylase to improve the production of dopamine in Escherichia coli.

Fuqiang Song; Wujiu Chen; Fengli WU; Xiaoshuang Wang; Fuping LU; Qinhong Wang

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Heterogeneous expression of DOPA decarboxylase to improve the production of dopamine in Escherichia coli.

Author: Fuqiang SONG ¹ ; Wujiu CHEN ² ; Fengli WU ² ; Xiaoshuang WANG ² ; Fuping LU ¹ ; Qinhong WANG ²
Author Information

1. Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin University of Science & Technology, Tianjin 300457, China.
2. CAS Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Science, Tianjin 300308, China.
Publication Type:Journal Article
Keywords: DOPA decarboxylase; Escherichia coli; L-DOPA; dopamine; metabolic engineering
MeSH: Animals; Dopa Decarboxylase/genetics*; Dopamine/biosynthesis*; Drosophila melanogaster/genetics*; Escherichia coli/metabolism*; Humans; Metabolic Engineering
From: Chinese Journal of Biotechnology 2021;37(12):4266-4276
CountryChina
Language:Chinese
Abstract: Dopamine is the precursor of a variety of natural antioxidant compounds. In the body, dopamine acts as a neurotransmitter that regulates a variety of physiological functions of the central nervous system. Thus, dopamine is used for the clinical treatment of various types of shock. Dopamine could be produced by engineered microbes, but with low efficiency. In this study, DOPA decarboxylase gene from Sus scrofa (Ssddc) was cloned into plasmids with different copy numbers, and transformed into a previously developed L-DOPA producing strain Escherichia coli T004. The resulted strain was capable of producing dopamine from glucose directly. To further improve the production of dopamine, a sequence-based homology alignment mining (SHAM) strategy was applied to screen more efficient DOPA decarboxylases, and five DOPA decarboxylase genes were selected from 100 candidates. In shake-flask fermentation, the DOPA decarboxylase gene from Homo sapiens (Hsddc) showed the highest dopamine production (3.33 g/L), while the DOPA decarboxylase gene from Drosophila Melanogaster (Dmddc) showed the least residual L-DOPA concentration (0.02 g/L). In 5 L fed-batch fermentations, production of dopamine by the two engineered strains reached 13.3 g/L and 16.2 g/L, respectively. The residual concentrations of L-DOPA were 0.45 g/L and 0.23 g/L, respectively. Finally, the Ssddc and Dmddc genes were integrated into the genome of E. coli T004 to obtain genetically stable dopamine-producing strains. In 5 L fed-batch fermentation, 17.7 g/L of dopamine was produced, which records the highest titer reported to date.