Engineering Lactococcus lactis Services for Live Biotherapeutics Drug Discovery

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One of the potential uses of recombinant Lactococcus lactis strains is the treatment of metabolic disorders such as diabetes, obesity, and auto-immune disorders such as atherosclerosis and encephalomyelitis. Creative Biolabs is one of the largest live biotherapeutics products (LBP) services suppliers. We have expertise and capabilities in providing high-quality LBP solutions.

Lactococcus Lactis: Heterologous Protein Delivery

Lactococcus lactis has made great progress from its initial discovery and application in dairy fermentation to the application of biotechnology in genetic engineering to produce various recombinant proteins and metabolites that transcend heterologous barriers. Key desirable characteristics of this Gram-positive lactic acid non-colonizing gut bacteria include its generally recognized safe (GRAS) status, probiotic properties, inclusion body and endotoxin-free, surface display and exocrine techniques, and multiple selections of cloning and induced expression vectors. This has made Lactococcus lactis a desirable and promising host on par with other well-established model bacterial or yeast systems.

Fig.1 The NICE system in L. lactis. (Frelet-Barrand, 2022)Fig.1 The Nisin-Controlled Gene Expression (NICE) system in L. lactis. (Frelet-Barrand, 2022)

Lactococcus lactis Genetic Engineering Services

Services
  • Gene knock-out
  • Gene knock-in/Gene insertion
Deliverables

Engineered strain in glycerol stock

Quality Control
  • Sequencing Validation
  • Western Blot Validation (Optional)
  • PAGE
  • or cell-based assay of your interest
Delivery Time

Starting from 4-6 weeks.

Materials from Clients
  • Overall construction scheme
  • Host strain
  • Target gene name
  • Target gene sequence, if the whole genome sequence is unavailable

Advantages

  • The growth of L. lactis is simple, the cost is low, and there are various genetic methods and vector systems.
  • The potential for host overexpression of homologous and allogenic proteins has been explored.
  • This bacteria can be studied directly with intact cells and membrane vesicles.

Case Studies

Case 1: Lactobacillus lactis Expression Strain Construction

Objective: Construction of Lactobacillus lactis strain expressing A enzyme

Result: Verified by SDS-PAGE and WB, the target protein was expressed in the secretion supernatant and lysed cell supernatant of Lactobacillus lactis NZ9000.

Method:

  1. Vector Construction (vector pNZ8148).
  2. Fusion Protein Expression (strain NZ9000).

Turnaround time: 6-8 weeks.

Deliverable:

  1. Strains
    1. Lactobacillus lactis NZ9000 A enzyme expression strain.
    2. A enzyme clone strain.
    3. Plasmid.
  2. Project report
    Including detailed materials and methods, PCR, and Western Blot validation results.

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Frequently Asked Questions

How have people benefited from Lactococcus lactis?

The most important function of L. lactis is to boost the immune system, and it is particularly effective in delivering antigens that stimulate mucosal immunity against respiratory pathogens.

Published Data

Recombinant Lactococcus lactis NZ3900 Expressing Human FGF21 Reduced Body Weight of Mice

FGF21 is a metabolic regulator that plays an important role in metabolic diseases such as obesity and diabetes. It is also expressed in mice, and the mouse source is highly homologous to human FGF21. In recent years, it has been widely studied as a potential drug target for the treatment of metabolic diseases. Scientists used Lactococcus lactis, a food-grade bacterium, as the host to express FGF21. In the NZ3900/PNZ8149 expression system, the LacF gene replaces the antibiotic resistance gene as the selective marker, which is safe and can reduce the antibiotic resistance crisis. The recombinant FGF21 expressing strain could express the human FGF21 well in the body and ameliorate obesity in Db/Db mice. Furthermore, a prolonged half-life of human FGF21 was achieved through oral gavage of the human FGF21-expressing strain. Therefore, oral human FGF21 may open up a new avenue for the treatment of obesity and related diseases.

Fig.3 Expression of FGF21 in the strain NZ3900. (Cao, 2020) Fig.2 Expression of recombinant Human FGF21 by Lactobacillus lactis NZ3900. (Cao, 2020)

As a common inducible host bacterium, Lactococcus lactis has an important role in research and application. Because of its potential in the production of heterologous proteins of therapeutic or technological value, Lactococcus lactis is becoming increasingly popular, among which the nisin-controlled gene expression (NICE) system is the most commonly used. In its combined form, Lactococcus lactis NZ3900/PNZ8149 is a crucial member of the NICE system. Creative Biolabs is a diverse, knowledgeable, and agile team, with a solid leadership universally recognized in the world of LBP. We have specialized in the custom services of LBP development for researchers around the world. If you are interested in our recombinant lactobacillus expression strain construction services, please contact us for more details.

References

  1. Frelet-Barrand, Annie. "Lactococcus lactis, an attractive cell factory for the expression of functional membrane proteins." Biomolecules 12.2 (2022): 180.
  2. Cao, W-Y., et al. "Recombinant Lactococcus lactis NZ3900 expressing bioactive human FGF21 reduced body weight of Db/Db mice through the activity of brown adipose tissue." Beneficial microbes 11.1 (2020): 67-78.

For Research Use Only. Not intended for use in food manufacturing or medical procedures (diagnostics or therapeutics). Do Not Use in Humans.

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For Research Use Only. Not intended for use in food manufacturing or medical procedures (diagnostics or therapeutics). Do Not Use in Humans.

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