Application of Adaptive Laboratory Evolution in Lipid and Terpenoid Production in Yeast and Microalgae

Abstract

Due to the complexity of metabolic and regulatory networks in microorganisms, it is difficult to obtain robust phenotypes through artificial rational design and genetic perturbation. Adaptive laboratory evolution (ALE) engineering plays an important role in the construction of stable microbial cell factories by simulating the natural evolution process and rapidly obtaining strains with stable traits through screening. This review summarizes the application of ALE technology in microbial breeding, describes the commonly used methods for ALE, and highlights the important applications of ALE technology in the production of lipids and terpenoids in yeast and microalgae. Overall, ALE technology provides a powerful tool for the construction of microbial cell factories, and it has been widely used in improving the level of target product synthesis, expanding the range of substrate utilization, and enhancing the tolerance of chassis cells. In addition, in order to improve the production of target compounds, ALE also employs environmental or nutritional stress strategies corresponding to the characteristics of different terpenoids, lipids, and strains.

Harnessing genetic engineering to drive economic bioproduct production in algae

“…Successful experiments have been conducted for the enhancement of metabolic pathways, bioproducts, increased uptake of specific substrates, tolerance to abiotic stress, and biofuel production to name a few ( LaPanse et al, 2021 ; Zhang et al, 2021 ). This has led to the increase of desirable natural products such as terpenoids, carotenoids, and lipids ( Fu et al, 2013 ; Arora et al, 2020 ; Jia et al, 2023 ). ALE can also be used in the reverse as a method to improve the degradation of particular compounds such as phenols, while still increasing biomass ( Wang et al, 2016 ).…”

Recent Advances and Multiple Strategies of Monoterpenoid Overproduction in Saccharomyces cerevisiae and Yarrowia lipolytica

“…66 This approach leverages the principles of natural selection, enabling strains to autonomously undergo genetic modifications in response to adaptive stress, thus securing survival and growth advantages in specified environments. 67 Here, ALE involves subjecting yeast to specific survival pressures to drive its evolution toward improved growth and production, a technique that has been successfully applied in both E. coli and yeasts. 68−71 Some approaches can achieve short-term ALE strategies to enhance limonene tolerance in Y. lipolytica.…”

“…Adaptive evolution (ALE) enables microorganisms to naturally change, via long-term cultivation under controlled laboratory conditions with specific selection pressures such as high temperatures, low pH, high salinity, or the presence of particular chemicals . This approach leverages the principles of natural selection, enabling strains to autonomously undergo genetic modifications in response to adaptive stress, thus securing survival and growth advantages in specified environments . Here, ALE involves subjecting yeast to specific survival pressures to drive its evolution toward improved growth and production, a technique that has been successfully applied in both E. coli and yeasts. − Some approaches can achieve short-term ALE strategies to enhance limonene tolerance in Y. lipolytica .…”

An Update on Microbial Biosynthesis of β-Caryophyllene, a Sesquiterpene with Multi-Pharmacological Properties

“…Retrosynthesis technologies will be supplemented with high-throughput procedures enabling the automated design of microbial strain engineering to aid in these efforts [54,[83][84][85]. Efforts will also be made to find more efficient terpene synthases [86], particularly in plants rich in BCP; to conduct membrane engineering for increased storage capacity [54,83,87,88]; to induce or overexpress efflux pumps, which have broad substrate specificity and export toxic metabolites outside the cells [85,89,90]; and to explore additional opportunities for applying the ALE technique [54,83,91,92]. In situ product extraction will receive more attention [54,83], especially since it has proven effective for improving the BCP titer in E. coli [19].…”