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Scylla paramamosain anti-bacterial polypeptide Sp-NPFin and its application

A technology of antibacterial peptides and pseudo-cave crabs, applied in the fields of application, antibacterial drugs, peptides, etc., can solve the problems of less research and achieve the effect of broad antibacterial spectrum, significant antibacterial effect, and fast sterilization rate

Active Publication Date: 2019-07-19
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still little research on the role of NPF in the immune response, especially in crustaceans

Method used

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  • Scylla paramamosain anti-bacterial polypeptide Sp-NPFin and its application
  • Scylla paramamosain anti-bacterial polypeptide Sp-NPFin and its application
  • Scylla paramamosain anti-bacterial polypeptide Sp-NPFin and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1 Preparation of the antibacterial polypeptide Sp-NPFin of pseudocave crab

[0036] The amino acid sequence of the antibacterial polypeptide Sp-NPFin of the pseudo-scyllas is:

[0037] Arg-Pro-Ser-Thr-Arg-Ser-Ala-Pro-Gly-Pro-Ala-Ser-His-Ile-Gln-Ala-Leu-Glu-Lys-Thr-Leu-Lys-Phe-Leu-Gln- Leu-Gln-Glu-Leu-Gly-Lys-Ile-Tyr-Ser-His-Met-Thr-Arg-Pro-Arg-Phe-Gly-Lys-Arg-Ser (SEQ ID NO: 01)

[0038] The antimicrobial peptide Sp-NPFin of Scylla sclerophyllus can be obtained with a purity of more than 95% by using the existing solid-phase chemical synthesis method. The antimicrobial polypeptide Sp-NPFin of Scylla pseudocavena in this example was synthesized by Jill Biochemical (Shanghai) Co., Ltd. by solid-phase synthesis method, and the molecular weight of the polypeptide, HPLC and other detection information were provided.

[0039] The physical and chemical parameters of the antibacterial peptide Sp-NPFin are shown in Table 1.

[0040] Table 1 Physicochemical paramete...

Embodiment 2

[0043] Example 2 Determination of the minimum inhibitory concentration (MIC: Minimum InhibitionConcentration) and the minimum bactericidal concentration (MBC: Minimum Bactericidal Concentration) of the antibacterial polypeptide Sp-NPFin of the crab crab

[0044] The strains involved in this example are: Staphylococcus aureus, Bacillus subtilis, Corynebacterium glutamicum, Bacillus cereus, Staphylococcus epidermidis (Staphylococcus epidermidis), Micrococcus lysodeikticus, Escherichia coli, Pseudomonas Aeruginosa, Pseudomonas stutzeri, Shigella flexneri Shigella flexneri, Pseudomonas fluorescens, Aeromonas hydrophila, Vibrio harveyi, Vibrio fluvialis, Vibrio alginolyticus (Vibrio alginolyticus), Vibrio parahaemolyticus, Candida albicans, Cryptococcus neoformans, Pichia pastoris, Fusarium graminearum, Fusarium oxysporum Fusarium oxysporum, Aspergillus fumigatus, Aspergillus ochraceus, Aspergillus niger, Fusarium solani. Pichia pastoris GS115 was purchased from Invitrogen, and oth...

Embodiment 3

[0060] Example 3 Scylla pseudocaveus antibacterial polypeptide Sp-NPFin bactericidal kinetic curve

[0061] Pseudomonas aeruginosa and Staphylococcus aureus were selected as the test bacteria, and the bactericidal kinetics of the antibacterial polypeptide Sp-NPFin of Scylla syringae were determined.

[0062] The specific method is similar to the antibacterial activity assay described in Example 2. Adjust the final concentration of Sp-NPFin to 1 times MBC (Pseudomonas aeruginosa: 12 μM, Staphylococcus aureus: 24 μM). At 10min, 30min, 60min, 120min, 180min, 360min after co-incubation, mix the blank control group, negative control group, and test group in the 96-well cell culture plate, draw 6 μL of Pseudomonas aeruginosa suspension and dilute to 600 μL In DPBS, after mixing, pipette 40 μL and spread it on a nutrient broth plate, and incubate it upside down at 37°C for 1-2 days, record the number of single clones of Pseudomonas aeruginosa, and calculate the bactericidal index. ...

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Abstract

The invention discloses a scylla paramamosain anti-bacterial polypeptide Sp-NPFin and its application. A molecular formula is C227H373N71O62S1, and its amino acid sequence is shown as SEQ ID NO: 01. The scylla paramamosain anti-bacterial polypeptide Sp-NPFin has strong antibacterial activity and antifungal activity, good antibacterial effect, wide antibacterial spectrum and fast sterilization rate. The scylla paramamosain anti-bacterial polypeptide Sp-NPFin is derived from crustaceans and can be applied to aquatic feed additives, can be developed to anti-mold preservatives, antibacterial agents and antibacterial drugs, and has broad application prospects.

Description

technical field [0001] The invention belongs to the technical field of marine molecular biology, and in particular relates to an antimicrobial polypeptide Sp-NPFin of Scylla pseudocaveus and an application thereof. Background technique [0002] Neuropeptides are a kind of neuroactive highly conserved multi-effect peptides released by the nervous system and various tissues, which widely exist in the nervous system of various organ levels from Hydra to humans. In both vertebrates and invertebrates, neuropeptides have very important regulatory functions and are very conserved. Some neuropeptides of invertebrates are very similar or homologous to those of vertebrates. [0003] Vertebrate neuropeptide Y (Neuropeptide Y, NPY), also known as neuropeptide tyrosine, was first isolated from pig brain by Swedish scientist Tatemoto et al. in 1982, and belongs to the pancreatic polypeptide family. In mammals, NPY has many functions such as promoting food intake, regulating body temperat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435A61K38/17A61P31/04A23K20/147A23K50/80
CPCA61K38/00A23K20/147A23K50/80A61P31/04C07K14/43509
Inventor 陈芳奕张华王克坚
Owner XIAMEN UNIV
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