5-Aminolevulinic Acid (5-ALA) Strain Engineering Service

5-Aminolevulinic Acid (5-ALA) is a vital biological precursor with broad applications in agricultural science as an environmentally friendly plant growth regulator, and in medicine as a photodynamic therapy sensitizer. However, the existing industrial bioproduction method suffers from low titer and high costs, severely limiting its wider application, especially in large-scale agriculture.

CD Biosynsis focuses on enhancing the performance of microbial hosts (such as Corynebacterium glutamicum and E. coli ) for 5-ALA synthesis. We employ core technologies like CRISPR-Cas9 precision editing to alleviate metabolic feedback inhibition and implement multi-strategy collaborative metabolic engineering. Our goal is to achieve a significant breakthrough in fermentation concentration and purity. We provide clients with advanced, cost-effective strain solutions to drive the industrialization and wider adoption of 5-ALA.

Pain Points

In current 5-ALA bioproduction, strain performance directly dictates economic viability. The critical bottlenecks are:

• Low Fermentation Titer: The typical biological fermentation concentration only reaches 20-25 g/L . This low yield necessitates higher processing volumes, directly contributing to elevated extraction costs and restricting price competitiveness in agricultural markets.
• Severe Metabolic Feedback Inhibition: The 5-ALA synthesis pathway is often tightly regulated by downstream products (like heme ), leading to strong feedback inhibition on key enzymes (such as C}_5$ pathway enzymes), which halts synthesis prematurely.
• Insufficient Product Purity: Fermentation processes often produce metabolic byproducts that are difficult to separate, resulting in a final product purity that is too low for pharmaceutical applications, and potentially affecting the efficacy of agricultural formulations.
• Imbalanced Precursor Supply: The supply of essential precursors (like succinyl-CoA and glycine) for 5-ALA synthesis is often inadequate or unbalanced within the host cell, limiting the overall flux towards the target product.

Overcoming the yield limitation requires precise metabolic pathway engineering to unlock the full production potential of the host strain.

Solutions

CD Biosynsis utilizes collaborative, multi-strategy engineering to address the core challenges of 5-ALA production, ensuring maximized yield and improved purity:

CRISPR-Cas9 Metabolic Inhibition Relief

           

We employ CRISPR-Cas9 to precisely modify or knock out regulatory regions of key synthesis pathway genes, effectively alleviating feedback inhibition caused by downstream products and extending the production phase.

5-ALA Pathway Enzyme Optimization

We screen highly efficient ALA synthase from diverse sources and optimize its expression level and enzyme activity, ensuring the entire pathway operates at its maximum flux rate towards 5-ALA.

Precursor Supply Enhancement

The metabolic pathways responsible for succinyl-CoA and glycine supply are engineered and reinforced, redirecting carbon flux away from competing pathways to ensure a robust supply of 5-ALA precursors.

Byproduct Pathway Knockout

Non-essential metabolic branches that divert carbon flux or generate impurities are systematically knocked out to minimize byproduct formation, significantly boosting final product purity and yield.

This multi-strategy approach is validated through controlled fermentation experiments, providing a reliable pathway for achieving high 5-ALA titers.

Advantages

Choosing CD Biosynsis's 5-ALA strain engineering service offers the following core value:

Titer Breakthrough Focus

Solutions are specifically engineered to dismantle the metabolic ceiling, targeting a fermentation titer significantly higher than the 20-25 g/L benchmark , directly impacting production cost.

Multi-Strategy Collaborative Optimization

We combine pathway amplification, inhibition relief, and precursor balancing, employing a synergistic strategy rather than single-point modification for robust and stable high-yield strains.

Purity Improvement through Design

By eliminating byproduct pathways upstream, we ensure the fermentation broth contains fewer impurities, simplifying downstream processing and achieving the high purity required for advanced applications.

Versatility in Host Systems

Our expertise covers common 5-ALA production strains, including E. coli , Corynebacterium glutamicum , and yeast, allowing for the selection of the most cost-effective and scalable host.

Data-Driven Metabolic Analysis

We employ advanced metabolic modeling before editing, ensuring the chosen targets are the most impactful for flux maximization , leading to higher success rates.

We are dedicated to providing stable, high-performance strains to accelerate the adoption of 5-ALA in both agricultural and pharmaceutical fields.

Process

CD Biosynsis's 5-ALA strain engineering service follows a standardized research workflow, ensuring every step is precise and controllable:

• Requirement Alignment and Baseline Analysis: Define the target titer/purity, and choose the optimal microbial host. Collect the strain's foundational data and conduct preliminary metabolic flux analysis to pinpoint rate-limiting steps.
• Technical Solution Design: Based on flux analysis, design gene editing targets for metabolic inhibition relief and precursor pathway reinforcement. Formulate a detailed Strain Engineering Research Protocol utilizing CRISPR-Cas9 or other tools.
• Strain Editing and Construction: Complete the construction of vectors, cell transformation, and high-throughput screening. Molecular verification confirms the precise editing of target genes, yielding the engineered high-yield 5-ALA strain.
• Performance Validation Experiments: Conduct comparative fermentation experiments under optimized conditions (e.g., fed-batch), measuring the difference in titer g/L ), yield g/g ), and purity between the engineered and wild-type strains.
• Result Report Output: Compile a Strain Engineering Experimental Report that includes fermentation curves, mass spectrometry-based purity analysis, and a technical summary, providing objective performance assessment and scale-up guidance.

Technical communication is maintained throughout the process, focusing on timely performance feedback and strategic adjustments to the metabolic engineering plan.

Achieve cost-effective 5-ALA production through specialized strain engineering! CD Biosynsis provides customized 5-ALA strain engineering solutions:

• Detailed Metabolic Flux Report analyzing the production bottlenecks of your current strain.
• Contracted clients receive discounts on fermentation optimization consultation , ensuring lab results translate effectively to industrial scale.
• Experimental reports include complete raw data on titer, yield, and purity , supporting regulatory and scale-up documentation.