Industrial Amylase Engineering Service for Enhanced Stability and Efficiency

Industrial Amylases are essential biocatalysts in starch liquefaction, brewing, and textile desizing. However, their industrial application is limited by poor stability at high temperatures (leading to high energy consumption) and a narrow pH adaptation range , requiring precise condition control. Furthermore, low catalytic efficiency necessitates higher enzyme loads, increasing production costs.

CD Biosynsis offers a comprehensive bioengineering solution focused on improving the performance and production of Amylase. Our core strategy involves the directed evolution of enzyme molecules to enhance thermal and pH stability and boost specific activity. This is combined with the optimization of the secretion system in Bacillus subtilis , an efficient industrial host, to maximize the final yield of the high-performance enzyme. This integrated approach aims to deliver a robust and cost-effective Amylase for large-scale industrial use.

Pain Points

Industrializing Amylase production and application faces these performance and host-related issues:

• Thermal Instability: Many native amylases lose activity rapidly above 70℃, complicating high-temperature starch processing and increasing cooling costs.
• Narrow pH Range: Most industrial processes require enzymes to be active across a broad pH range, but existing enzymes often have a narrow pH adaptation range , leading to efficiency loss.
• Low Catalytic Efficiency: Lower specific activity (kcat) means higher enzyme dosage is required per batch, directly increasing operational costs.
• Host Secretion Bottlenecks: While Bacillus subtilis is a strong secretor, challenges such as protease degradation and inefficient signal peptide cleavage still limit final functional enzyme yields.

A successful solution must dramatically improve the enzyme's intrinsic robustness and its yield from the host.

Solutions

CD Biosynsis employs enzyme engineering and host optimization for developing superior Amylase:

Directed Evolution of Enzyme Molecules

           

We use error-prone PCR and DNA shuffling coupled with high-throughput screening to generate variants with enhanced thermostability and pH tolerance.

Optimization of the Secretion System in Bacillus subtilis

We engineer the signal recognition particle (SRP) pathway components and optimize the signal peptide to maximize the efficient export of the target Amylase.

Rational Enzyme Design and Stabilization

Based on structural analysis, we introduce stabilizing mutations (e.g., disulfide bridges or salt bridges ) to enhance the enzyme's structural rigidity at extreme temperatures.

Host Protease Knockout for Stability

Genes encoding major extracellular proteases in B. subtilis are deleted to minimize the degradation of the valuable secreted Amylase during fermentation.

This systematic approach is focused on enhancing both the intrinsic performance and the bioproduction yield of the Amylase.

Advantages

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

High Thermostability

The engineered enzyme is intended to maintain high activity at elevated temperatures, potentially leading to reduced energy usage in industrial processes.

Broad pH Adaptability

Directed evolution aims to produce an Amylase variant that operates efficiently across a wider industrial pH range , increasing versatility.

Enhanced Catalytic Efficiency

Improvements in specific activity aim to reduce the required enzyme dosage per batch, directly lowering raw material costs.

Maximized Secretion Yield

Optimization of the host's secretion system is focused on significantly increasing the final enzyme output in the fermentation medium.

Simplified Purification Potential

Using the highly secretory Bacillus subtilis host, combined with protease knockout, provides a cleaner broth , potentially simplifying downstream steps.

We provide a comprehensive platform aimed at overcoming the stability and cost challenges of industrial enzyme application.

Process

Our Industrial Amylase engineering service follows a standardized, iterative research workflow:

• Enzyme Characterization: Determine the wild-type Amylase's initial activity, T_{50 (thermal stability), and optimal pH range , setting baseline targets for improvement.
• Directed Evolution Strategy: Generate diverse mutant libraries using methods like ITCHY or random mutagenesis and implement a high-throughput screening assay for enhanced stability/efficiency.
• Host Secretion Optimization: Engineer the B. subtilis host by optimizing promoter strength, signal peptide sequences , and potentially knocking out competing genes for high secretion.
• Protease and Genome Modification: Systematically delete major extracellular protease genes in the B. subtilis genome to prevent product degradation.
• Scale-Up and Fermentation Validation: Test the final engineered strain/enzyme combination in fed-batch fermentation to assess yield, cost-efficiency, and long-term stability under simulated industrial conditions.
• Result Report Output: Compile a detailed Experimental Report including mutant sequence data, specific activity metrics (kcat/Km), thermostability profiles, and final fermentation titer , supporting technology transfer.

Technical communication is maintained throughout the process, focusing on timely feedback regarding enzyme performance and host productivity.

Explore the potential for robust, cost-effective industrial enzyme production. CD Biosynsis provides customized strain and enzyme engineering solutions:

• Detailed Kinetic and Stability Analysis Report , demonstrating the improvement in T_{50 and kcat/Km values.
• Consultation on fermentation and purification strategies optimized for the engineered B. subtilis strain.
• Experimental reports include complete raw data on enzyme yield, thermal deactivation rates, and pH activity profiles , essential for commercial application assessment.