Due to the toxic side effects of traditional antibiotics and the emergence of resistant strains, efforts are being made to find alternatives to antibiotics. Defensin, a novel bioactive peptide with a broad antibacterial spectrum, has a completely different mechanism of action than antibiotics that block macromolecular biosynthesis. Defensin can rapidly kill broad-spectrum pathogenic microorganisms, and as an active substance of the organism itself, it is relatively non-immunogenic, with fewer bacteria resistant to it and pathogenic bacteria less likely to develop resistance to it, so it can replace antibiotics to play a broad-spectrum and efficient antibacterial effect in agriculture and animal husbandry.
Defensin preparation
Defensins can be obtained mainly through three routes, extraction from cells or body fluids, chemical synthesis, and recombinant expression and purification. However, the expression of defensins in tissues is very small and the purification process is difficult and costly, so genetic engineering preparation is undoubtedly the preferred method for the mass production of defensins. Lifeasible uses advanced genetic engineering technology to transform defensin genes into yeast for large-scale recombinant expression and fermentation after molecular modification, which can greatly improve the content and activity of defensins and effectively deal with pathogenic microorganisms. Of course, we also offer prokaryotic expression and baculovirus expression systems for customers to choose from.
Purification of recombinant defensins
- Capture phase. We collect the target proteins from the supernatant of bacterial cell breakage, including body lysate and other systems, and remove some of the tramp proteins. For unlabeled proteins, we use ion exchange chromatography, and for labeled proteins, we use affinity chromatography or ion exchange chromatography followed by affinity chromatography.
- Crude purification stage. We recover the target peptide from enzymatic or chemical lysis of fusion protein system, yeast fermentation supernatant, animal cell culture supernatant, and other solutions. This step is mainly performed by cation exchange chromatography or gel chromatography. The dissociation of the target peptide from the fusion protein is a key step in the purification.
- Refinement stage. We mainly use reversed-phase chromatography to obtain high-purity target peptides.
For different sources of recombinant defensins, we will design a purification strategy with fewer steps and lower costs to obtain high recovery according to the biochemical properties of various proteins in the system.
Defensins in livestock
The use of antibiotic additives seriously disrupts the micro-ecological balance of animal intestines, and drug residues affect the quality of livestock products and human health. Since defensins are peptide components, they are easily degraded to amino acids by proteases in the body, and there are generally no residues in the body after animals have consumed them. Moreover, defensins from mammals have a small relative molecular mass and are thermally stable and water-soluble, so they can be absorbed in the intestinal tract of animals.
We can use genetic engineering methods to produce environmentally friendly defensin feed additives or add them to diets. This can regulate the expression of defensin genes and achieve their effects, such as reducing stillbirths, increasing newborn weight, and birth uniformity, improving weaning survival, reducing lactation morbidity, reducing stress, and improving subfertility and reproductive disorders. In addition, it can improve daily weight gain, reduce the feed-to-meat ratio, improve immunosuppression, improve immune response and make antibody levels more uniform.
Defensin in agriculture
We build defensin bioreactors for mass production and purification of defensin protein to help you use defensin in crop disease resistance breeding research to develop new resistant varieties. We have been able to transfer rabbit defensin NP-1 into wheat plants, and the results of pest and disease resistance tests show that wheat has improved resistance to powdery mildew, leaf rust, and stripe rust. The results of disease resistance in tomato plants obtained by constructing rabbit defensin NP-1 gene into plant expression vector showed that it was resistant to tomato cyanobacteria, which provides a new idea for breeding tomatoes for disease resistance.
With the development of biotechnology, the emergence of new defensin genes, and the continuous research on the defensin family, Lifeasible's highly efficient expression systems and bioreactors are constantly being updated, and we are committed to promoting defensins as a new generation of antimicrobial drugs widely used in agricultural and livestock production, creating more value for human beings. Please feel free to contact us to formulate your requirements.
For research use only, not intended for any clinical use.
