Terpenoids are a class of natural products widely distributed in living organisms, with isoprene as their fundamental structural unit. However, traditional plant extraction and chemical synthesis methods are often limited by low product purity, difficult separation, and complex synthetic steps, making it challenging to meet the demands of large-scale production. Conventional hosts such as Escherichia coli and Saccharomyces cerevisiae are utilized for terpenoid synthesis due to their advantages of short growth cycles and controllable cultivation conditions. Nevertheless, the complexity of terpenoid biosynthetic pathways poses significant challenges for these hosts in producing structurally complex terpenoids. In contrast, microalgae as photosynthetic microorganisms, possess well-developed endogenous terpenoid metabolic pathways, abundant precursor pools, and subcellular structures and regulatory mechanisms similar to those of plants, demonstrating significant advantages in the heterologous production of complex terpenoids. This review systematically summarizes recent advances in the production of heterologously synthesized terpenoids in eukaryotic microalgae, ranging from monoterpenes to triterpenes, and provides an in-depth analysis of key engineering strategies, including MEP/MVA pathway regulation, gene expression optimization, subcellular compartmentalization, and cultivation process intensification. In addition, the application potential of advanced tools such as CRISPR/Cas, microalgae-microorganism co-culture, and artificial intelligence is introduced. Finally, the major bottlenecks faced by microalgae as a sustainable green cell factory for terpenoid production are briefly analyzed, and future research directions are proposed.
This review summarizes recent advances in monoterpenoid biosynthesis, multilevel regulation, and heterologous production, with particular emphasis on major bottlenecks and optimization strategies for sustainable and efficient biomanufacturing.
Jun-Chi Zhang, Jiale Cui, Shang Li et al.· Natural Products and Biopros...· 0 citations
Artificial intelligence (AI) and machine learning (ML) have emerged as powerful tools for metabolic engineering by enabling pathway prediction, metabolic flux optimization, enzyme engineering, and identification of bottlenecks throughout terpenoid biosynthesis.
Aakash Kamalesan, K. Kumar, Bharathi Nathan et al.· Antonie van Leeuwenhoek· 0 citations
Terpenoids (isoprenoids) constitute one of the largest and most structurally diverse families of natural products with application ranging from flavors, pharmaceuticals to biofuels. Conventional extraction from plants is often limited by low yields, seasonal variability, and environmental constraints; whereas, chemical synthesis requires toxic chemicals and energy-intensive processes. Consequently, microbial cell factories have emerged as sustainable and industrially scalable alternatives for terpenoid biosynthesis. Different microorganisms possess distinct physiological and metabolic advantages, including efficient precursor supply, tolerance to toxic products, internal storage for hydrophobic compounds, utilization of renewable carbon sources, and compatibility with complex biosynthetic pathways. Furthermore, several microbial hosts have strains with Generally Recognized as Safe (GRAS) status, making them attractive candidates for food and nutraceutical applications, although their regulatory acceptance ultimately depends on the production strain, genetic modifications, manufacturing process, product, and intended use. In addition, photosynthetic cyanobacteria offer a promising platform for direct conversion of CO2 into terpenoids, providing opportunities for more resource-efficient and sustainable biomanufacturing. Therefore, strategic host selection is a crucial step in designing efficient microbial platforms for terpenoid production. The present review provides a host-centric perspective by comparing conventional and emerging microbial cell factories, highlighting their physiological strengths, product spectrum, industrial applicability, and strategic considerations for sustainable and application-specific terpenoid biomanufacturing.
Vibha Shukla, Virendra Shukla, Shweta Rawat et al.· Frontiers in Bioengineering...· 0 citations
Triterpenoids are important natural secondary metabolites with diverse bioactivities, including antioxidant, anti-inflammatory, and anti-cancer properties, making them valuable for applications in the pharmaceutical, cosmetic, and food industries. Currently, triterpenoids are mainly obtained through natural extraction or chemical synthesis. However, these conventional approaches are often limited by production efficiency, environmental burdens, and product diversity. Rapid advances in metabolic engineering and synthetic biology have promoted the emergence of heterologous biosynthesis as a promising, efficient, and sustainable strategy for triterpenoids production. In this review, we first summarize the classification and bioactive properties of triterpenoids, together with the challenges and potential solutions associated with their microbial synthesis. Then, we analyze the key characteristics of microbial hosts and their corresponding biosynthetic pathways for triterpenoids production, aiming to establish programmable platforms that overcome the limitations of natural biosynthesis. Subsequently, we propose metabolic engineering and synthetic biology strategies, including enzyme optimization, pathway optimization, compartmentalization engineering, and systems biology approaches, for optimizing matter and energy transmission and thereby enhancing triterpenoids production. We further discuss the potential challenges for scaling up triterpenoids production from laboratory-scale studies to industrial-scale applications, including the optimization of large-scale fermentation process and the improvement of downstream extraction and recovery. Finally, we discuss the techno-economic feasibility and industrial prospects of microbial triterpenoid production, highlight current regulation and governance in synthetic biology related to triterpenoids biosynthesis, analyze existing limitations, and propose potential solutions to provide insights for future research on the biomanufacturing of triterpenoids.
Ning Jiao, Zhanpeng Shan, Xiu-Lai Chen· Biotechnology Advances· 0 citations
The heterologous production of terpene in microbial hosts is often limited by inefficient and unstable pathway expression, creating a major bottleneck for industrial-scale synthesis. While E. coli as a chassis offers significant advantages, such as rapid growth, ease of cultivation, and genetic tractability. Its endogenous supply of terpenoid precursors remains a critical constraint, fundamentally restricting high-yield production. To address this challenge, we developed a genomically integrated Mevalonate (MVA) pathway from Actinomycetota in E. coli BL21(DE3) to enhance terpene precursor supply. Our approach began with an in silico multi-layer global genome mining analysis of 25,261 Actinomycetota genomes to identify a series of MVA pathway enzymes with potentially high catalytic efficiency, created a high-efficiency chassis E. coli MVA platform (ecMVA-1 and ecMVA-2) for terpene precursor synthesis. Its functionality was validated by testing eight distinct TSs. Among them, the fermentation of artemisinin precursor amorphadiene using a 5-liter bioreactor yielded 947.80 mg/L. These results indicated that E. coli (MVA) is well-suited for TS studies in the laboratory as well as holding significant promise for industrial applications. In addition, this in silico approach offers a new perspective for metabolic engineering and provides potential reservoir of diverse chassis for the industrial production of terpenoid-derived compounds.
Wenchao Liu, Xueying Tian, W. Wong et al.· Metabolic Engineering· 0 citations
