Recent advances in high-throughput ¹³C-fluxomics

Area of Science:

• Synthetic biology
• Metabolic engineering
• Biotechnology

Background:

• Synthetic biology enables the creation of numerous microbial cell factories through high throughput (HT) strain engineering.
• Analyzing the metabolic state, specifically intracellular fluxes, of these engineered strains is a critical bottleneck in process development.
• Current high throughput (HT) tools exist for cultivation, sampling, isotopic analysis, and data processing, but integrated HT ¹³C-fluxomics workflows are not yet established.

Purpose of the Study:

• To review recent advancements in high throughput (HT) ¹³C-fluxomics.
• To identify and highlight the key bottlenecks hindering the widespread adoption of HT ¹³C-fluxomics.
• To discuss the requirements for establishing true HT ¹³C-fluxomics workflows in metabolic engineering.

Main Methods:

• Review of recent literature and technological developments in high throughput (HT) ¹³C-fluxomics.
• Analysis of existing HT tools across different stages of the ¹³C-fluxomics pipeline.
• Identification of integration challenges and bottlenecks in current workflows.

Main Results:

• Significant progress has been made in individual HT tools for cultivation, sampling, isotopic analysis, and data processing.
• A lack of integration and standardization across these HT tools prevents the establishment of seamless HT ¹³C-fluxomics workflows.
• Key bottlenecks include the need for automated sampling, faster isotopic analysis, and integrated software for flux calculation.

Conclusions:

• Overcoming current bottlenecks in cultivation, analytics, and data processing is essential for realizing the potential of HT ¹³C-fluxomics.
• Further development and integration of HT tools are required to enable rapid metabolic state analysis of engineered microbial strains.
• The emergence of true HT ¹³C-fluxomics workflows will accelerate synthetic biology and metabolic engineering efforts.