Ethyl Lactate Bioproduction Engineering Service

Ethyl Lactate (EL) is an environmentally friendly solvent, flavor agent, and pharmaceutical intermediate. Its utility stems from its low toxicity and biodegradability. Conventional production via chemical esterification of lactic acid and ethanol suffers from insufficient purity in chemical synthesis due to side reactions and difficulty in removing residual reactants/catalysts. Biosynthetic routes in yeast often face low fermentation yield , primarily because the primary fermentation product of yeast is ethanol (competing with lactic acid) and the esterification step itself is inefficient.

CD Biosynsis offers a synthetic biology service focused on the integrated, high-titer production of Ethyl Lactate from sugars in Saccharomyces cerevisiae. Our core strategy involves modification of ethanol metabolism pathway in Saccharomyces cerevisiae . We engineer yeast to reroute carbon flux from ethanol (the competing product) to lactic acid and ethanol precursors, ensuring both alcohol and acid components are available. This includes deleting the major alcohol dehydrogenase gene (ADH1) and introducing a highly efficient lactate dehydrogenase (LDH) gene. Critically, this is coupled with overexpression of ester synthase . We introduce and heavily overexpress a highly active, solvent-tolerant ester synthase (e.g., a specific lipase or esterase) to efficiently catalyze the final in situ esterification of bio-derived lactic acid and ethanol/butanol into Ethyl Lactate. This integrated approach aims to deliver a high-yield, high-purity, bio-based Ethyl Lactate product, simplifying downstream purification.

Pain Points

Developing a competitive biosynthetic route for Ethyl Lactate faces these key challenges:

• Low Fermentation Yield: Wild-type yeast prefers to produce ethanol over lactic acid. The inherent competition for carbon flux severely limits the yield of the lactic acid precursor.
• Insufficient Purity in Chemical Synthesis: Chemical esterification often requires harsh catalysts and high heat, resulting in residual impurities (e.g., catalyst traces, unreacted acid/alcohol) that complicate large-scale pharmaceutical or food-grade applications.
• Lack of Native Esterase: Yeast lacks an endogenous enzyme that can efficiently catalyze the formation of Ethyl Lactate from the two bio-derived components at a commercially viable rate.
• Product Toxicity: Ethyl Lactate, being a solvent, can be toxic to the host organism (S. cerevisiae) at high concentrations, limiting the final achievable titer.

A successful solution must reroute the central carbon metabolism away from ethanol, toward lactic acid, and introduce a highly active, dedicated ester synthase.

Solutions

CD Biosynsis utilizes advanced metabolic and enzyme engineering to optimize EL production in S. cerevisiae:

Modification of Ethanol Metabolism Pathway in S. cerevisiae

           

We implement a knockout of ADH1 (Alcohol Dehydrogenase) and a co-overexpression of heterologous LDH (Lactate Dehydrogenase) to divert carbon flux toward lactic acid and away from ethanol.

Overexpression of Ester Synthase

We introduce a heterologous, high-performance ester synthase/lipase and overexpress it using strong, constitutive promoters to ensure efficient, rapid conversion of lactic acid and ethanol to EL.

Fine-tuning Redox Balance

We co-optimize $\text{NADH}/\text{NAD}^+$ balance through metabolic shunts to ensure sufficient reducing power for the engineered lactate pathway, improving overall carbon yield .

In Situ Product Recovery (ISPR) Integration

We integrate a two-phase fermentation system or membrane separation to continuously remove the toxic EL product , mitigating toxicity and increasing final titer.

This systematic approach is focused on establishing a highly directed carbon flux toward both precursors and maximizing the efficiency of the final enzymatic coupling.

Advantages

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

High Integrated Titer

Rerouting the central carbon metabolism and overexpressing the final enzyme leads to maximal EL concentration in the fermenter.

Bio-based High Purity Product

Enzymatic synthesis avoids harsh catalysts, resulting in a cleaner product profile suitable for food and pharmaceutical applications.

Sustainable Production Route

Fermentation utilizes renewable sugar feedstock , reducing dependence on petrochemicals. [Image of Cost Reduction Icon]

Simplified Purification

ISPR and high purity synthesis simplify downstream recovery and reduce refining costs .

Robust Yeast Host

Saccharomyces cerevisiae is a well-established industrial workhorse, providing a robust and scalable platform .

We provide a sustainable and cost-effective biosynthetic platform for industrial Ethyl Lactate production.

Process

Our Ethyl Lactate strain engineering service follows a rigorous, multi-stage research workflow:

• Metabolic Flux Rerouting: Knockout the ADH1 gene and implement the heterologous LDH gene to direct carbon from pyruvate toward lactic acid.
• Ester Synthase Optimization: Introduce and overexpress a highly active ester synthase (e.g., from Candida or Rhizomucor) under a strong, constitutive promoter.
• Ethanol/Lactic Acid Balance Tuning: Fine-tune the pathway to ensure stoichiometrically sufficient amounts of both the acid and alcohol components for the esterification reaction.
• ISPR System Development: Design and integrate a compatible In Situ Product Recovery (ISPR) method (e.g., membrane or solvent extraction) to manage EL toxicity.
• Fermentation Performance Validation: Test the final engineered strain in fed-batch fermentation to assess EL titer, yield, and purity .
• Result Report Output: Compile a detailed Experimental Report including gene modification data, enzyme kinetics, and fermentation metrics (yield, titer, and purity) , supporting commercial scale-up.

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

Explore the potential for a high-titer, bio-based Ethyl Lactate supply. CD Biosynsis provides customized strain and enzyme engineering solutions:

• Detailed Titer and Yield Analysis Report , demonstrating success in carbon flux redirection toward EL.
• Consultation on optimized ISPR method design and downstream purification strategies.
• Experimental reports include complete raw data on carbon yield (g EL/g sugar) and final product purity (e.g., residual acid/alcohol levels).