Pathway Engineering for Phenethylamine Production in Escherichia coli

Abstract

In this study, the metabolic pathway of phenethylamine synthesis was reconstructed by chromosomal integration and overexpression of the Enterococcus faecium pdc gene encoding phenylalanine decarboxylase in Escherichia coli. The genes encoding 3deoxy-D-arabinoheptulosonate-7-phosphate synthase (aroG), shikimate kinase II (aroL), chorismate mutase/prephenate dehydratase (pheA), and tyrosine aminotransferase (tyrB) in the phenethylamine synthetic pathway were sequentially chromosomally overexpressed. The phosphotransferase system was replaced by deleting the ptsH-ptsI-crr genes and chromosomally overexpressing the genes encoding galactose permease (galP) and glucokinase (glk). In addition, the zwf gene encoding glucose-6-phosphate dehydrogenase in the pentose phosphate pathway was chromosomally overexpressed, generating the final engineered E. coli strain AUD9. The AUD9 strain produced 2.65 g L −1 phenethylamine with a yield of 0.27 g of phenethylamine g −1 glucose in batch fermentation; fed-batch fermentation of AUD9 produced 38.82 g L −1 phenethylamine with a productivity of 1.08 g L −1 h −1 phenethylamine, demonstrating its potential for industrial fermentative production of phenethylamine.

High-titre production of aromatic amines in metabolically engineered Escherichia coli

“…As depicted in Figure 5b, the maximum phenethylamine production of 555 ± 50 mg/L was achieved by strain PEA3.0 after 72 h, which was increased by 2.28‐fold compared with that of strain PEA1.0 (169 ± 20 mg/L). The phenethylamine level achieved in the shake flask by our study was 1.2 times higher than that of a recent report (Xu & Zhang, 2020). In addition, we found that both strain PEA1.0 and PEA3.0 gave a similar growth profile (Figure 5b).…”

“…Heterologous expression of tyrosinase and decarboxylase in the L‐tyrosine overproducing E. coli achieved 260 mg/L dopamine (Nakagawa et al, 2011). The metabolic pathway for the production of phenethylamine in E. coli was rebuilt using chromosomal integration of the phenethylamine synthesis pathway and the PP pathway, and the engineered strain produced 460 mg/L of phenylethylamine in a shake flask and 38.82 g/L in fed‐batch fermentation (Xu & Zhang, 2020).…”

High-titer production of aromatic amines in metabolically engineered Escherichia coli

“…5b, the maximum phenethylamine production of 555±50 mg/L was achieved by strain PEA3.0 after 72 h, which was increased by 2.28-fold compared with that of strain PEA1.0 (169±20 mg/L). The phenethylamine level achieved in the shake flask by our study was also much higher than that of a recent report (Xu and Zhang 2020). In addition, we found that both strain PEA1.0 and PEA3.0 gave a similar growth profile (Fig.…”

“…Finally, 1.965±0.205 g/L tyramine, 703±21 mg/L dopamine, and 555±50 mg/L phenethylamine were obtained from 40 g/L glycerol in shake flask cultivation. The titers achieved by us were much higher than previous reports under shake flask conditions (Koma et al 2012; Nakagawa et al 2011; Xu and Zhang 2020).…”

“…The copyright holder for this preprint this version posted January 14, 2022. ; https://doi.org/10.1101/2022.01.14.476304 doi: bioRxiv preprint higher than previous reports under shake flask conditions (Koma et al 2012;Nakagawa et al 2011;Xu and Zhang 2020).…”