Nisin Engineering Service for Enhanced Yield and Spectrum

Nisin is a widely used bacteriocin in the food industry, acting as a natural preservative (E number E234). Its application is currently limited by a low fermentation yield in the natural host ( Lactococcus lactis ) and a narrow antibacterial spectrum , primarily effective against Gram-positive bacteria. Expanding its functionality and increasing its cost-effectiveness are key goals for wider adoption.

CD Biosynsis offers a synthetic biology service focused on optimizing the production and function of Nisin. Our core strategy involves the metabolic regulation modification of Lactococcus lactis to enhance the flux of precursor amino acids and reduce competing metabolic pathways, thereby increasing Nisin synthesis. This is combined with domain engineering of the Nisin peptide to introduce modifications that may expand the antibacterial spectrum, including efficacy against some Gram-negative pathogens. This integrated approach aims to deliver a high-yield, broad-spectrum Nisin variant for the food and pharmaceutical sectors.

Pain Points

Improving the industrial viability of Nisin requires addressing these production and functional limitations:

• Low Fermentation Yield: The native producer, L. lactis , often has low growth rates and low specific productivity , resulting in a high fermentation cost per unit of Nisin.
• Narrow Antibacterial Spectrum: Nisin acts primarily on Gram-positive bacteria by targeting lipid II. Its inability to penetrate the outer membrane of Gram-negative bacteria severely restricts its applications.
• Biosynthesis Bottlenecks: The complex post-translational modifications (dehydration, cyclization) required to form active Nisin can be rate-limiting steps in the L. lactis host.
• Self-Immunity System: The host cell's immunity system (e.g., NisI and NisFEG) limits the maximum amount of Nisin that can be safely produced without toxicity.

A successful solution must maximize the metabolic efficiency and engineer the peptide for expanded functionality.

Solutions

CD Biosynsis utilizes advanced synthetic biology to enhance Nisin production and function:

Metabolic Regulation Modification of Lactococcus lactis

           

We employ genome editing to enhance the flux of key amino acid precursors (e.g., Serine and Threonine) and optimize energy metabolism to support higher synthesis rates.

Domain Engineering to Expand the Antibacterial Spectrum

We modify the Nisin peptide structure (e.g., N-terminus or specific hinge regions) to potentially increase hydrophobicity or charge, enabling it to better penetrate the outer membrane of Gram-negative bacteria.

Biosynthesis Machinery Optimization

We overexpress or modify the enzymes responsible for post-translational modification (e.g., NisB dehydratase, NisC cyclase) to eliminate folding and processing bottlenecks.

Self-Immunity System Tuning

We employ controlled expression of immunity genes to increase the host's tolerance to higher internal Nisin concentrations, supporting higher total production yield.

This systematic approach is focused on overcoming both the fermentation efficiency issues and the functional limitations of the natural Nisin peptide.

Advantages

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

High Fermentation Titer

Metabolic and regulatory modifications aim to significantly boost the specific productivity (mg}/\text{L}/\text{h) of the host strain.

Expanded Antibacterial Spectrum

Engineered variants are focused on showing efficacy against previously resistant Gram-negative bacteria , expanding market potential.

Enhanced Biosynthesis Efficiency

Optimization of post-translational machinery aims to ensure a higher conversion rate of the precursor peptide into active Nisin.

Natural and Safe Preservative

As a naturally derived bacteriocin, Nisin is a Generally Recognized as Safe (GRAS) substance, meeting strong consumer demand for clean labels.

Cost Reduction Potential

Higher fermentation yield is focused on reducing the overall cost per unit of active Nisin, improving competitiveness. [Image of Cost Reduction Icon]

We provide a biosynthetic platform aimed at maximizing the yield and functional spectrum of Nisin production.

Process

Our Nisin strain engineering service follows a standardized, iterative research workflow:

• Metabolic Pathway Analysis: Map the L. lactis genome to identify bottlenecks in amino acid synthesis and energy flow that limit Nisin production.
• Biosynthesis Gene Optimization: Overexpress or rationally engineer the post-translational modification enzymes (NisB/NisC) and transport system (NisT) for efficient precursor processing.
• Nisin Domain Engineering: Use rational design or directed evolution to introduce mutations in the Nisin structural gene to test for expanded Gram-negative activity .
• Immunity System Tuning: Engineer the Nisin immunity system (NisFEG) to allow the host to tolerate higher product concentrations in the medium.
• Fermentation Performance Validation: Test the final engineered strain in fed-batch fermentation to assess volumetric productivity and final Nisin titer .
• Result Report Output: Compile a detailed Experimental Report including genetic modification data, Nisin titer, and antibacterial spectrum testing (including MIC against Gram-negative strains), supporting product registration.

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

Explore the potential for a high-performance, broad-spectrum Nisin supply. CD Biosynsis provides customized strain and peptide engineering solutions:

• Detailed Titer and Spectrum Analysis Report , demonstrating the success of yield enhancement and functional expansion.
• Consultation on fermentation strategies optimized for Nisin production in the L. lactis host.
• Experimental reports include complete raw data on Minimal Inhibitory Concentration (MIC) against target bacteria and final product purity, essential for quality control.