Biodiesel (Fatty Acid Methyl Ester, FAME) Engineering Service

Biodiesel, typically in the form of Fatty Acid Methyl Esters (FAME), is a leading candidate for renewable liquid transportation fuel. Its commercialization is significantly challenged by the high cost of raw materials (traditional vegetable oils or animal fats) and the low efficiency of the ester exchange (transesterification) reaction when using enzymatic catalysts.

CD Biosynsis offers a synthetic biology service focused on both microbial feedstock generation and process efficiency. Our core strategy involves the modification of oil-producing yeast lipid synthesis pathway to maximize the yield of affordable, microbial oil (single-cell oil) as a direct replacement for costly traditional feedstocks. This is combined with the directed evolution of lipase enzymes to optimize their activity, solvent tolerance, and thermal stability, significantly improving the efficiency of the transesterification step. This integrated approach aims to deliver an economically competitive and environmentally sustainable bioproduction platform for FAME.

Pain Points

The economic viability of biobased FAME is constrained by these major hurdles:

• High Raw Material Cost: Traditional feedstock oils account for a significant portion of the final biodiesel price , making it difficult to compete with low-cost fossil diesel.
• Low Efficiency of Ester Exchange: Chemical transesterification requires harsh conditions. Enzymatic conversion using lipase is gentler but often suffers from low catalytic rate and poor tolerance to the methanol/oil mixture.
• Lipid Synthesis Bottlenecks: Native oleaginous microbes often divert carbon flux to non-lipid growth products, resulting in a lower overall oil content in the biomass.
• Glycerol Byproduct Issues: The transesterification process generates glycerol as a major byproduct, which requires costly purification and disposal or valorization.

A cost-effective solution requires both a cheaper, high-yield feedstock and a more efficient biocatalytic conversion process.

Solutions

CD Biosynsis addresses feedstock and conversion challenges through metabolic and enzyme engineering:

Modification of Oil-Producing Yeast Lipid Synthesis Pathway

           

We employ CRISPR and genome editing to upregulate key lipid accumulation enzymes (DGA1) and downregulate competing pathways, maximizing TAG (Triacylglycerol) yield from low-cost carbon sources.

Directed Evolution of Lipase for Optimization

We use directed evolution to enhance the lipase biocatalyst, focusing on increased solvent (methanol) tolerance , thermal stability, and higher specific activity for FAME production.

Glycerol Metabolism Engineering

We modify the yeast's central metabolism to enable the conversion of the glycerol byproduct into valuable products or back into the FAME synthesis pathway, improving process economics.

Simultaneous Oil Synthesis and Transesterification

We explore whole-cell catalysis by engineering the oleaginous yeast to express the optimized lipase on its surface or within lipid bodies, enabling single-step conversion.

This systematic approach is focused on lowering feedstock costs through microbial oil and improving the efficiency of the enzymatic conversion process.

Advantages

Our Biodiesel engineering service is dedicated to pursuing the following production goals:

Affordable Microbial Feedstock

Microbial oil production decouples FAME price from volatile commodity vegetable oil markets, providing a low-cost, sustainable alternative .

Enhanced Transesterification Rate

Optimized lipase and whole-cell catalysts are focused on achieving a faster, more complete conversion of TAG to FAME.

Improved Biocatalyst Robustness

Engineered lipases exhibit higher tolerance to methanol and higher temperatures , making the enzymatic process more industrial-friendly.

Sustainable Production Route

Biocatalysis is a non-toxic, non-corrosive process that reduces chemical waste compared to conventional chemical conversion.

Cost Reduction Potential

Reduced feedstock cost and improved conversion efficiency are focused on achieving economic parity with fossil fuels . [Image of Cost Reduction Icon]

We provide an integrated platform aimed at maximizing the yield and minimizing the cost of FAME production.

Process

Our Biodiesel engineering service follows a standardized, integrated research workflow:

• Feedstock Strain Engineering: Modify the lipid synthesis pathway of an oleaginous yeast to maximize TAG accumulation (>70\% cell dry weight) from simple carbon sources.
• Lipase Directed Evolution: Generate mutant libraries of lipase and screen for variants with high specific activity and increased tolerance to methanol and co-solvents.
• Lipase Expression System: Develop a high-yield expression system for the engineered lipase, focusing on surface display or encapsulation for whole-cell catalysis.
• Glycerol Byproduct Valorization: Engineer the host's metabolism to convert glycerol into a valuable coproduct or back into cell mass, eliminating waste stream issues.
• Process Integration and Validation: Test the engineered biocatalyst/whole cell in a transesterification reaction with microbial oil, measuring FAME yield and purity .
• Result Report Output: Compile a detailed Experimental Report including strain modification data, lipase kinetics, and final FAME yield, purity, and process economics , supporting industrial scale-up.

Technical communication is maintained throughout the process, focusing on timely feedback regarding lipid yield and conversion efficiency.

Explore the potential for a cost-competitive, sustainable Biodiesel supply. CD Biosynsis provides customized strain and enzyme engineering solutions:

• Detailed Lipid Yield and Composition Analysis Report , confirming the quantity and quality of the microbial oil feedstock.
• Consultation on process optimization for whole-cell transesterification, including methanol staging and temperature control.
• Experimental reports include complete raw data on lipase activity (U}/\text{mg), methanol tolerance, and FAME conversion efficiency , essential for process design.