Pathway engineering for high-yield production of lutein in Escherichia coli

Abstract

Lutein is an industrially important carotenoid pigment, which is essential for photoprotection and photosynthesis in plants. It is crucial for maintaining human health due to its protective ability from ocular diseases. However, its pathway engineering research has scarcely been performed for microbial production using heterologous hosts, such as Escherichia coli, since the engineering of multiple genes are required there. These genes, which include tricky key carotenoid biosynthesis genes typically derived from plants, encode two sorts of cyclases (lycopene ε- and β-cyclase) and cytochrome P450 CYP97C. In this study, upstream genes effective for the increase in carotenoid amounts, such as isopentenyl diphosphate (IPP) isomerase (IDI) gene, were integrated into the E. coli JM101 (DE3) genome. The most efficient set of the key genes (MpLCYe, MpLCYb, and MpCYP97C) was selected from among corresponding genes derived from various plant (or bacterial) species using E. coli that had accumulated carotenoid substrates. Furthermore, to optimize the production of lutein in E. coli, we introduced several sorts of plasmids that contained some of the multiple genes into the genome-inserted strain and compared lutein productivity. Finally, we achieved 11 mg/L as lutein yield at a mini jar level. Here, high-yield production of lutein was successfully performed using E. coli through approaches of pathway engineering. The findings obtained here should be a base reference for substantial lutein production with microorganisms in the future.

Lycopene Production in Dedicated Novel Chasses for Lignocellulosic Waste Material Utilisation Capable of Sustained Coculture

“…The plasmids pACCRT-EIB 14 and pACHP-LYC 41 were obtained as a kind gift of Prof Norihiko Misawa. pACCRT-EIB encodes the genes crtE, crtB and crtI from Agrobacterium auranauticans .…”

“…Cells were cultured overnight in the relevant medium, according to the previous investigators 14,15,41 . Lycopene was quantified according to methods of Kim and Keasling 13 .…”

Marine Microbial-Derived Resource Exploration: Uncovering the Hidden Potential of Marine Carotenoids

“…Carotene production in microbes can be increased by providing stimulants and optimizing environmental conditions during the cultivation process. The addition of mevalonic acid as a precursor was found to increase carotene formation by 120% in R. mucilaginosa and 35% in R. glutinis [102].…”

Microalgal lutein biosynthesis: Recent trends and challenges to enhance the lutein content in microalgal cell factories

“…It is very difficult to produce lutein in non-photosynthetic microorganisms through heterologous expression due to challenges involved in lycopene asymmetric cyclization. However, some studies have succeeded in the heterologous biosynthesis in non-photosynthetic microorganisms such as Escherichia coli (Takemura et al, 2021) and Saccharomyces cerevisiae (Bian et al, 2021). Integration of several genes including isopentenyl diphosphate isomerase (IDI), lycopene ϵ-cyclase (LCY-e), lycopene b-cyclase (LCY-b) and cytochrome P450 97C (CYP97C) in E. coli leads to the synthesis of 11 mg/L of lutein (Takemura et al, 2021).…”

“…However, some studies have succeeded in the heterologous biosynthesis in non-photosynthetic microorganisms such as Escherichia coli (Takemura et al, 2021) and Saccharomyces cerevisiae (Bian et al, 2021). Integration of several genes including isopentenyl diphosphate isomerase (IDI), lycopene ϵ-cyclase (LCY-e), lycopene b-cyclase (LCY-b) and cytochrome P450 97C (CYP97C) in E. coli leads to the synthesis of 11 mg/L of lutein (Takemura et al, 2021). In the other approach, combinatorial engineering in S. cerevisiae provides lutein content up to 438 mg/g CDW (Bian et al, 2021).…”