Carotenoids are a class of terpenes of commercial interest and exert important biological functions. Engineering morphological and biosynthetic aspects of Escherichia coli cell membrane could improve its storage capacity for β-carotene. However, how the synthesis of phosphatidylethanolamine, the major component of the cell membrane, was not discussed in detail. In this work, the synthesis of phosphatidylethanolamine was divided into three modules to discuss their synergetic effect, by expressing in different combinations. Overexpressing the upstream module 1 in CAR016 caused a 30.5% increase of β-carotene specific production (from 10.1 mg/g to 13.7 mg/g DCW); combined overexpressing module 1 and module 2 in CAR016 led to a 122% increase of β-carotene specific production (from 10.5 mg/g to 22.3 mg/g DCW). The optimal expression combination of the phosphatidylethanolamine synthetic pathway was obtained, which further increased the content of the cell membrane for β-carotene storage, and improved its production. The membrane engineering strategy opens up a new direction for engineering microbial producers for a large spectrum of hydrophobic molecules.

As an important model industrial microorganism, Escherichia coli has been widely used in pharmaceutical, chemical industry and agriculture. In the past 30 years, a variety of new strategies and techniques, including artificial intelligence, gene editing, metabolic pathway assembly, and dynamic regulation have been used to design, construct, and optimize E. coli cell factories, which remarkably improved the efficiency for biotechnological production of chemicals. In this review, three key aspects for constructing E. coli cell factories, including pathway design, pathway assembly and regulation, and optimization of global cellular performance, are summarized. The technologies that have played important roles in metabolic engineering of E. coli, as well as their future applications, are discussed.
Artificial Intelligence ; Escherichia coli/genetics* ; Gene Editing ; Metabolic Engineering ; Metabolic Networks and Pathways/genetics*