Compartmentalization and transporter engineering strategies for terpenoid synthesis

Abstract

Microbial cell factories for terpenoid synthesis form a less expensive and more environment-friendly approach than chemical synthesis and extraction, and are thus being regarded as mainstream research recently. Organelle compartmentalization for terpenoid synthesis has received much attention from researchers owing to the diverse physiochemical characteristics of organelles. In this review, we first systematically summarized various compartmentalization strategies utilized in terpenoid production, mainly plant terpenoids, which can provide catalytic reactions with sufficient intermediates and a suitable environment, while bypassing competing metabolic pathways. In addition, because of the limited storage capacity of cells, strategies used for the expansion of specific organelle membranes were discussed. Next, transporter engineering strategies to overcome the cytotoxic effects of terpenoid accumulation were analyzed. Finally, we discussed the future perspectives of compartmentalization and transporter engineering strategies, with the hope of providing theoretical guidance for designing and constructing cell factories for the purpose of terpenoid production.

Keywords: Compartmentalization; Metabolic engineering; Terpenoids; Transporter.

Fig. 1

a ER, b LD, c peroxisome and d mitochondrion used for compartmentalization strategies. G3P: glycerol-3-phosphate; PA: phosphatidic acid; DAG: diacylglycerol; TAG: triacylglycerol; SE: sterol esters; PLN1: perilipin; PAH1: phosphatidic acid phosphatase; DGA1: diacylglycerol acyltransferase

Fig. 2

Schematic representation of membrane engineering strategies using E. coli as an example. The overexpression of membrane-bending proteins and the enhancement of membrane lipids will lead to membrane expansion for more terpenoid storage. G3P: glycerol-3-phosphate; Diacylglycerol-3P: diacylglycerol-3-phosphate; Tsr: chemotaxis receptor protein; PlsB: glycerol-3-phosphate acyltransferase; PlsC: 1-acylglycerol- phosphate acyltransferase

Fig. 3

Schematic representation of transport system engineering strategies using S. cerevisiae and E. coli as an example. a Using ABC transporters in S. cerevisiae for terpenoid secretion. b Engineering E. coli cells to produce more outer membrane vesicles by reducing the crosslinks between the inner and outer membrane, and enhancing certain membrane components

Fig. 4

Future prospects of strategies that can effectively promote terpenoid synthesis using S. cerevisiae as an example. a Peptides with dual-localization may target enzyme complex to two different organelles. b Using methods like AlphaFold and molecular dynamics simulation for protein engineering to increase the substrate specificity of ABC transporters. c Mining terpenoid transporters to accelerate intracellular transportation