Engineered mitochondrial production of monoterpenes in Saccharomyces cerevisiae

Yeast cell factories, particularly Saccharomyces cerevisiae, have proven valuable for the synthesis of non-native compounds, ranging from commodity chemicals to complex natural products. One significant challenge has been ensuring sufficient carbon flux to the desired product. Traditionally, this has been addressed by strategies involving “pushing” and “pulling” the carbon flux toward the products by overexpression while “blocking” competing pathways via downregulation or gene deletion. Colocalization of enzymes is an alternate and complementary metabolic engineering strategy to control flux and increase pathway efficiency toward the synthesis of non-native products. Spatially controlling the pathway enzymes of interest, and thus positioning them in close proximity, increases the likelihood of reaction along that pathway. This mini-review focuses on the recent developments and applications of colocalization strategies, including enzyme scaffolding, construction of synthetic organelles, and organelle targeting, in both S. cerevisiae and non-conventional yeast hosts. Challenges with these techniques and future directions will also be discussed.

Section: Organelle Targetingmentioning

confidence: 99%

Successful Enzyme Colocalization Strategies in Yeast for Increased Synthesis of Non-native Products

Yocum

Pham

Silva

2021

“…By transplanting the whole FPP pathway together with amorpha-4,11-diene synthase into yeast mitochondria, the yield of amorpha-4,11-diene in engineered strain reached 427 mg/L (Yuan and Ching, 2016). Yee et al (2019) targeted the geraniol biosynthetic pathway to the S. cerevisiae mitochondria to protect the GPP pool from consumption by the cytosolic ergosterol pathway. The production of geraniol in mitochondrial was 6-fold increase compared to cytosolic producing strains ( Figure 1A).…”

Section: Subcellular Engineering and Cell Free Systemmentioning

confidence: 99%

Production of Terpenoids by Synthetic Biology Approaches

Zhang

Hong

2020

Terpenoids are a large family of natural products with remarkable diverse biological functions, and have a wide range of applications as pharmaceuticals, flavors, pigments, and biofuels. Synthetic biology is presenting possibilities for sustainable and efficient production of high value-added terpenoids in engineered microbial cell factories, using Escherichia coli and Saccharomyces cerevisiae which are identified as well-known industrial workhorses. They also provide a promising alternative to produce non-native terpenes on account of available genetic tools in metabolic engineering and genome editing. In this review, we summarize the recent development in terpenoids production by synthetic biology approaches.

“…The concentration of acetyl-CoA in mitochondria is 20-30 times higher than that in the cytoplasm (Galdieri et al, 2014). Many studies focused on the mitochondrial acetyl-CoA pool to promote the production of PNPs such as amorpha-4,11-diene (Yuan and Ching, 2016), geraniol, 8-hydroxygeraniol, and nepetalactol (Yee et al, 2019). Recently, dual engineering of metabolic pathways in the cytoplasm and organelles was performed for high-level PNPs production.…”

Section: Subcellular Compartmentalization Of Metabolic Pathwaysmentioning

confidence: 99%

Microbial Cell Factory for Efficiently Synthesizing Plant Natural Products via Optimizing the Location and Adaptation of Pathway on Genome Scale

Yang

Feng

2020

Plant natural products (PNPs) possess important pharmacological activities and are widely used in cosmetics, health care products, and as food additives. Currently, most PNPs are mainly extracted from cultivated plants, and the yield is limited by the long growth cycle, climate change and complex processing steps, which makes the process unsustainable. However, the complex structure of PNPs significantly reduces the efficiency of chemical synthesis. With the development of metabolic engineering and synthetic biology, heterologous biosynthesis of PNPs in microbial cell factories offers an attractive alternative. Based on the in-depth mining and analysis of genome and transcriptome data, the biosynthetic pathways of a number of natural products have been successfully elucidated, which lays the crucial foundation for heterologous production. However, there are several problems in the microbial synthesis of PNPs, including toxicity of intermediates, low enzyme activity, multiple auxotrophic dependence, and uncontrollable metabolic network. Although various metabolic engineering strategies have been developed to solve these problems, optimizing the location and adaptation of pathways on the whole-genome scale is an important strategy in microorganisms. From this perspective, this review introduces the application of CRISPR/Cas9 in editing PNPs biosynthesis pathways in model microorganisms, the influences of pathway location, and the approaches for optimizing the adaptation between metabolic pathways and chassis hosts for facilitating PNPs biosynthesis.