Levan Engineering Service

Levan, a polyfructan, is an exopolysaccharide with applications as a Prebiotics/Film Formers in food, cosmetics, and pharmaceuticals due to its water-solubility and unique structure. The current production methods face a major hurdle: Low yield and high cost of purification from traditional Bacillus fermentation. This limits its cost-effectiveness and scalability for industrial use.

CD Biosynsis offers a two-pronged strategy focused on enzyme and feedstock efficiency: Enzyme Engineering: Overexpress the Levansucrase enzyme in E. coli or yeast and optimize the enzyme reaction to control the degree of polymerization DP. This ensures a high-quality product with tailored properties. Additionally, we implement Feedstock Utilization: Engineer the host to utilize cheap feedstocks like sucrose. By optimizing the use of economical carbon sources, we significantly reduce the overall production cost.

Pain Points

The industrial production of Levan faces these primary constraints:

• Low Titer and Yield: The traditional Bacillus fermentation often results in low Levan titer compared to biomass production, making recovery inefficient.
• High Purification Cost: Bacillus fermentation produces Levan in a complex broth with cell debris and proteins, requiring extensive and costly purification steps e.g. alcohol precipitation.
• Uncontrolled DP: The Degree of Polymerization DP is critical for Levan functionality e.g. low DP for prebiotics, high DP for films. Traditional methods struggle to precisely control this molecular weight distribution.
• Feedstock Constraints: Levansucrase requires sucrose, which can be a relatively expensive feedstock compared to simple glucose or glycerol.

A cost-effective solution must improve Levansucrase activity and enable cheaper, high-titer production.

Solutions

CD Biosynsis utilizes enzyme engineering and metabolic optimization for enhanced Levan production:

High-Yield Enzyme Expression

           

We overexpress the Levansucrase enzyme in robust, high-density fermentation hosts e.g. E. coli or yeast for high enzyme titer and purity.

Precise DP Control

We optimize enzyme reaction conditions e.g. sucrose concentration and use enzyme mutants to precisely tune the Degree of Polymerization DP of the Levan product.

Cost-Effective Feedstock Use

We engineer the host Bacillus or E. coli to efficiently utilize cheap sucrose sources e.g. molasses and co-utilize other sugars, reducing raw material costs.

Simplified Downstream Processing

By using cell-free or extracellular enzyme systems in a clean host, we significantly simplify the purification steps required for high-purity Levan.

This biological and process optimization strategy ensures a high-quality, cost-competitive Levan supply.

Advantages

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

Tunable Molecular Weight DP Icon

Enzyme optimization allows for precise control over the DP, tailoring Levan for prebiotic or material applications.

High Enzyme Activity and Stability Icon

Overexpression in robust hosts provides large amounts of highly active Levansucrase, improving reaction kinetics.

Lower Feedstock Cost Icon

Host engineering enables the efficient use of cheap sucrose-based byproducts e.g. molasses, cutting production expenses.

Simplified Purification Icon

Cell-free or extracellular production dramatically reduces the steps and cost associated with downstream processing.

High Product Purity Icon

The in-vitro or enzyme reaction approach minimizes impurities related to biomass and other byproducts.

We deliver a scalable, high-purity, and functional Levan product for demanding industrial markets.

Process

Our Levan engineering service follows a rigorous, multi-stage research workflow:

• Levansucrase Gene Optimization: Clone Levansucrase genes from Bacillus or other sources and codon-optimize them for high expression in E. coli or yeast.
• Enzyme Production and Purification: Develop high-titer fermentation protocols for the recombinant Levansucrase host and establish a cost-effective purification method.
• DP Tuning Reaction Development: Optimize the extracellular enzymatic reaction by adjusting sucrose concentration, reaction time, and temperature to achieve target DP.
• Feedstock Metabolic Engineering: Engineer Bacillus or E. coli to efficiently take up and utilize low-cost sucrose-rich media e.g. molasses.
• Product Characterization and Validation: Determine Levan yield and analyze the molecular weight DP distribution e.g. via SEC-HPLC to confirm compliance with specifications.

Technical communication is maintained throughout the process, focusing on timely feedback regarding yield and product quality attributes.

Explore the potential for a cost-efficient, molecularly-defined Levan supply. CD Biosynsis provides customized strain and process engineering solutions:

• Detailed Levan Titer and DP Reports g/L, average DP from optimized runs.
• Consultation on extracellular reaction scale-up and enzyme recycling strategies.
• Experimental reports include complete raw data on Levansucrase activity, substrate conversion rates, and product rheology.