Inducible Lactococcus Expression Service

Lactococcus lactis has recently been identified as an effective gram-positive cell factory for the production of recombinant protein. As an expert in the biotechnology industry, Creative BioMart has now established a Nisin induced NICE (Nisin controlled gene expression system) system based lactic acid bacteria expression platform. We can provide a variety of expression schemes according to customer needs, including heat-induced expression, food grade expression and secretory expression. We promise to provide our customers with high supernatant, high purity, high activity of Lactobacillus protein expression services.

NICE Technology Description

Nisin is a natural bioactive antimicrobial peptide composed of 34 amino acids. It is the only bacteriocin allowed to be used as a food additive in the world.

An effective NICE system consists of three parts:

• Gram-positive host bacteria carrying nisR and nisK gene
• Nisin or its analogs and mutants as inducing molecules
• The plasmids containing nisA or nisF promoter fragments and MCS

Service Content

Expression procedure and delivery standard of L.lactis constitutive expression system.

Selection of expression system (if the customer provides the expression strain, this step will be omitted. Only the optional host strain of expression plasmid will be provided)
Available expression option	Standard constitutive expression system Food grade constitutive expression system
Construction of expression vector	Deliverable
Gene optimization Subcloning design Synthetic gene Subcloning to vector	1. Gene synthesis report 2. Sequencing validation report 3. Cloning strain containing expression plasmid (optional) 4. Expression plasmid 5. Plasmid instructions
Analysis and identification of protein expression test
Electroconversion Expression test Expression analysis and identification Expand scale	1. Positive expression strains 2. Small test expression process report 3. Try to express and analyze original images 4. Small test expression summary
Protein expression and purification
Protein affinity purification (extracellular or intracellular) Identify purified proteins using WB PFM detection (optional) MS detection (optional)	1. Protein purification process report 2. Original picture of protein purification 3. Delivery of protein 4. Western blot (labeled antibody) 5. Protein fingerprint (optional) 6. Molecular weight of protein mass spectrometry (optional)
Scale-up culture in fermentor
Amplification Culture (10-20l) Amplification Culture (above 50L)	1. Protein purity identification report 2. Finished protein (agreed price according to the results of expression test)

Why Choose NICE Expression System

• Expression of membrane proteins
• Secretion of proteins into the medium
• Less endogenous and no exogenous proteases
• Endotoxin-free food-grade expression system
• No spores
• Tightly controlled gene expression allows the production of toxic proteins
• Simple fermentation, scale-up and downstream processing. It is also suitable for bacteria other than lactic acid bacteria

What You Can Do With The Technology

• Overexpression of homologous and heterologous genes
• Application of industrial-scale protein production
• Expression of membrane proteins
• Protein secretion and surface anchoring
• Expression and analysis of toxic or essential gene products

Customers who need our assistance are more than welcome to contact us or submit an online inquiry. Our professional team will provide you with detailed guidance and solution ASAP.