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Scylla paramamosain antibacterial polypeptide Sp-LECin and application thereof

A technology of antibacterial peptides and pseudo-cave crabs, applied in the fields of application, antibacterial drugs, peptides, etc., can solve the problem of finding a small number of new antibacterial peptides, achieve strong antibacterial activity and antifungal activity, good antibacterial effect, and sterilization rate fast effect

Active Publication Date: 2022-02-08
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, most of the antimicrobial peptides included in the antimicrobial peptide library are obtained based on the analysis of the reported antimicrobial peptide sequences, EST fragments or structural domains, and the discovery of new antimicrobial peptides is still a minority.

Method used

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  • Scylla paramamosain antibacterial polypeptide Sp-LECin and application thereof
  • Scylla paramamosain antibacterial polypeptide Sp-LECin and application thereof
  • Scylla paramamosain antibacterial polypeptide Sp-LECin and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Preparation of the antibacterial polypeptide Sp-LECin from pseudocave crab

[0042] The amino acid sequence of the antibacterial polypeptide Sp-LECin of the pseudo-scyllas is:

[0043]Gly-Cys-Val-Phe-Leu-Leu-Pro-Ala-Lys-Pro-His-Asn-Tyr-Lys-Lys-Val-Phe-Leu-Ser-Lys-Gly-Val (SEQ ID NO.1)

[0044] The antibacterial polypeptide Sp-LECin of Scylla syringae can be obtained with a purity of more than 95% by using the existing solid-phase chemical synthesis method. GenScript (Jiangsu) Co., Ltd. was entrusted with GenScript (Jiangsu) Co., Ltd. to synthesize the antimicrobial polypeptide Sp-LECin of Scylla pseudocarpa in this example by solid-phase synthesis method, and provided detection information such as molecular weight of the polypeptide and HPLC.

[0045] The physical and chemical parameters of the antibacterial peptide Sp-LECin are shown in Table 1.

[0046] Table 1 Physicochemical parameters of antibacterial peptide Sp-LECin

[0047]

[0048] It can be see...

Embodiment 2

[0049] 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-LECin of the crab crab

[0050] The bacterial strains involved in this embodiment include: Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecium, Enterococcus faecalis, Listeria monocytogenes Listeria monocytogenes, Escherichia coli, Pseudomonas aeruginosa, Pseudomonas stutzeri, Shigella flexneri, Pseudomonas fluorescens Pseudomonas fluorescens, Acinetobacter baumannii, Fusarium oxysporum, Aspergillus niger. The strains were purchased from the Culture Collection Center of the Institute of Microbiology, Chinese Academy of Sciences, and stored in our laboratory.

[0051] The specific method is as follows:

[0052] (1) The preserved Staphylococcus aureus, drug-resistant Staphylococcus aureus, Staphylococcus epidermidis, Listeria mono...

Embodiment 3

[0065] Example 3 Scylla pseudocaveus antibacterial polypeptide Sp-LECin bactericidal kinetic curve

[0066] Enterococcus faecium and Acinetobacter baumannii were selected as the tested bacteria to measure the bactericidal kinetics of the antibacterial peptide Sp-LECin from Scylla pseudosclera.

[0067] The specific method is similar to the antibacterial activity assay described in Example 2. Adjust the final concentration of Sp-LECin to 1 times MBC (Enterococcus faecium: 24 μM; Acinetobacter baumannii: 12 μM). 10min, 20min, 30min, 45min, 60min, 90min, 120min after co-incubation, mix the blank control group, negative control group, and test group in the 96-well cell culture plate, and absorb 6 μL of Enterococcus faecium suspension to dilute to 600 μL In DPBS, after mixing, draw 40 μL and spread it on a nutrient broth plate, incubate it upside down at 37°C for 1-2 days, record the number of single colonies of Enterococcus faecium, and calculate the bactericidal index. At 5min,...

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Abstract

The invention discloses a scylla paramamosain antibacterial polypeptide Sp-LECin and an application thereof. The molecular formula of the antibacterial polypeptide Sp-LECin is C117H185N29O26S1, and the amino acid sequence of the antibacterial polypeptide Sp-LECin is as shown in SEQ ID NO. 1. The scylla paramamosain antibacterial polypeptide Sp-LECin has antibacterial activity and antifungal activity, is good in antibacterial effect, wide in antibacterial spectrum and high in sterilization rate, is derived from crustacean, can be applied to aquatic feed additives, can also be developed into preservatives, antibacterial mildew inhibitors, antibacterial drugs and the like, and has wide application prospects.

Description

technical field [0001] The invention belongs to the technical field of marine molecular biology, and specifically relates to polypeptides and applications thereof. Background technique [0002] Due to the abuse of antibiotics in the fields of medical and health care and aquaculture, the global problem of antibiotic pollution is becoming more and more serious. It is urgent to develop new therapeutic drugs that can resist drug-resistant bacterial infections and replace antibiotics. Antimicrobial Peptide (AMP) is an important innate immune factor present in various organisms. It has immune properties such as anti-bacteria, fungi, viruses and parasites. Because it is different from traditional antibiotics, it is not easy to produce drug resistance. Sex, known as "natural antibiotics" by scientists. According to the different charge properties, they are divided into cationic antimicrobial peptides and anionic antimicrobial peptides. Most antimicrobial peptides are cationic antim...

Claims

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Application Information

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IPC IPC(8): C07K14/435A01N37/46A01P1/00A01P3/00A61K38/17A61P31/04A23L3/3526A23K50/80A23K20/147
CPCC07K14/43509A01N37/46A61P31/04A23L3/3526A23K50/80A23K20/147A61K38/00A23V2002/00A23V2200/10A23V2250/55
Inventor 陈芳奕邱婉蕾王克坚彭会
Owner XIAMEN UNIV
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