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

Inactive Publication Date: 2018-11-06
NORTHWESTERN POLYTECHNICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

To solve the problems existing in the use of existing antibiotics and the existing selenium additives in food and feed have strong toxicity, poor water solubility, low bioavailability, large environmental pollution, and the existing production and preparation technology process is complicated, the production cost is high, There are problems such as unsafe factors

Method used

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  • Multifunctional compound microecological preparation nano-selenium-recombination expression VIP (vasoactive intestinal peptide)-lactococcus lactis and preparation method
  • Multifunctional compound microecological preparation nano-selenium-recombination expression VIP (vasoactive intestinal peptide)-lactococcus lactis and preparation method
  • Multifunctional compound microecological preparation nano-selenium-recombination expression VIP (vasoactive intestinal peptide)-lactococcus lactis and preparation method

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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...

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Abstract

The invention relates to multifunctional compound microecological preparation nano-selenium-recombination expression VIP (vasoactive intestinal peptide)-lactococcus lactis and a preparation method. Toxic sodium selenite can be converted into non-toxic red simple-substance nano-selenium, and synthesized nano-selenium is enriched in lactococcus lactis NZ9000 in cells. The invention also discloses amethod for preparing a multifunctional compound microecological preparation SENPs-rVIP-L.lactis NZ9000 based on lactococcus lactis NZ9000. The problems that existing selenium supplementing additives have high toxicity action and low bioavailability and easily cause environment pollution, existing production technical procedures are complicated, the cycle is long and the production cost is high andthe problems about remaining of antibiotics, drug resistance and the like are solved.

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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/74C12P3/00C12P21/02C12R1/46
CPCC12N15/74C12P3/00C12P21/02C07K14/47Y02A50/30
Inventor 徐春兰郭宇乔磊马丽程忆忆
Owner NORTHWESTERN POLYTECHNICAL UNIV
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