Multifunctional compound microecological preparation nano-selenium-recombination expression VIP (vasoactive intestinal peptide)-lactococcus lactis and preparation method
A technology of vasoactive intestinal peptide and compound microecology, which is applied in the fields of bioengineering and nanomaterial preparation, can solve the problems of strong toxicity of selenium additives, complex technical process, and low bioavailability, so as to improve intestinal health and wide application Prospect, high bioavailability effect
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Publication Date
- 2018-11-06
- Estimated Expiration
- Not applicable · inactive patent
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Abstract
Description
technical field
[0001] The invention belongs to the technical field of bioengineering and nano-material preparation, and relates to a multifunctional compound microecological preparation nano-selenium-recombinantly expressed vasoactive intestinal peptide-Lactococcus lactis and a preparation method. Background technique
[0002] The overuse of feed antibiotics leads to serious problems of drug residue and drug resistance. Therefore, it is urgent to reduce or even ban the use of antibiotics and find suitable substitutes. In recent years, many countries in the world have introduced corresponding policies to control the use of antibiotics. However, due to the rapid development of the breeding industry, the increase in breeding density and the increased risk of breeding animal diseases, there is an urgent need for new green feed additive products that can replace antibiotics. Today, green, safe and residue-free new green feed additives - probiotic microecological preparations ar...
Examples
Embodiment
[0048] Method for constructing secretory recombinant expression plasmid of antimicrobial peptide VIP
[0049] (1) Design and optimization of antimicrobial peptide VIP gene sequence:
[0050] use Lactococcus lactis expression system, since the pNZ8148 plasmid itself is not a secretory expression plasmid, in order to realize the secretion and expression of the antimicrobial peptide VIP in Lactococcus lactis, the signal peptide sequence SP was introduced when designing the VIP gene sequence usp45 , in order to facilitate the subsequent separation and purification of antimicrobial peptides, a His tag recognition sequence (HHHHH) was added to the nucleotide sequence of the antimicrobial peptides; KpnI and NcoI restriction endonucleases were used to construct recombinant plasmids, and sequences corresponding to pNZ8148 were designed at both ends of the gene The KpnI and NcoI enzyme cutting sites in the middle have the same sequence, and the nucleotide sequence of the newly designe...
Embodiment 6
[0068] Example 6: Antioxidant activity of Lactococcus lactis L.lactis NZ9000 enriched in nano-selenium
[0069] by establishing 500 µM H 2 o 2 The induced oxidative damage model of porcine intestinal epithelial cells IPEC-J2 cells was used to study the antioxidant activity of the microecological preparation prepared by the present invention. IPEC-J2 cells in the logarithmic growth phase were co-cultured with Lactococcus lactis NZ9000 enriched in nano-selenium (4 μg / mL selenium) for 8 h, and then the cells were exposed to 500 μM H 2 o 2 After further culturing for 12 h in serum-free medium, live cells were stained with Hoechst 33342 staining kit, and cell apoptosis was detected with Annexin V-FITC PI staining kit. The level of malondialdehyde (MDA) and the activity of total superoxide dismutase (T-SOD) in the cell culture medium were detected with corresponding kits. The results are attached Figure 4 As shown, Lactococcus lactis NZ9000 enriched with nano-selenium signific...