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CN-121991196-A - Small molecular peptide for inhibiting development of blue crab ovary and application thereof

CN121991196ACN 121991196 ACN121991196 ACN 121991196ACN-121991196-A

Abstract

The invention belongs to the technical field of biology, and particularly relates to a small molecular peptide for inhibiting the development of blue crab ovary and application thereof. The invention predicts the binding conformation according to the vitellogenin and the receptor sequence thereof to screen the optimal binding conformation, screens the hot spot residue according to the optimal binding conformation, calculates the binding energy change before and after the mutation of the hot spot residue, and determines the competitive inhibition polypeptide sequence. The competitive inhibition polypeptide can utilize the specificity of specific recognition of the blue crab vitellogenin and the receptor, can obviously slow down the development speed of blue crab ovary and effectively induce the formation of yellow crab, and can produce yellow crab products in a planned way.

Inventors

  • XU LIKUN
  • QIN JIAO
  • LUO MINGHAO
  • YANG MIN
  • JIANG XIAOSONG
  • ZHOU DONGPING
  • ZHAO XIAOYI
  • SUN WENJING

Assignees

  • 南方海洋科学与工程广东省实验室(湛江)

Dates

Publication Date
20260508
Application Date
20251219

Claims (10)

  1. 1. A small molecule peptide for inhibiting the development of blue crab ovary, which is characterized in that: the amino acid sequence of the small molecule peptide is as described in any one of a 1) to a 3): a1)KSLGNMGVMTPEAAD; a2)AADKVILYMEKENKKV; a3)KENKKVSNRVAAAQAFR。
  2. 2. The small molecule peptide of claim 1, wherein: The small molecule peptides include pharmaceutically acceptable salt forms thereof.
  3. 3. A fusion protein, characterized in that: the fusion protein comprises the small molecule peptide of claim 1 and optionally a tag sequence that facilitates expression and/or purification.
  4. 4. A biomaterial related to the small molecule peptide of any one of claims 1-2, the fusion protein of claim 3, the biomaterial comprising at least one of b 1) -b 11): b1 A nucleic acid molecule encoding the small molecule peptide of any one of claims 1-2 or the fusion protein of claim 3; b2 An expression cassette comprising b 1) the nucleic acid molecule; b3 A vector comprising b 1) the nucleic acid molecule; b4 A vector comprising b 2) said expression cassette; b5 A transgenic cell line comprising b 1) said nucleic acid molecule; b6 A transgenic cell line comprising b 2) said expression cassette; b7 A transgenic cell line comprising b 3) the vector; b8 A transgenic cell line comprising b 4) the vector; b9 A microorganism comprising b 1) said nucleic acid molecule; b10 A microorganism comprising b 2) said expression cassette; b11 A microorganism comprising b 3) said vector.
  5. 5. The small molecule peptide of any one of claims 1 to 2, the method for producing the fusion protein of claim 3, comprising any one of c 1) to c 2): c1 Expression and purification using the biomaterial of claim 4; c2 Is synthesized by using a chemical solid phase peptide synthesis method.
  6. 6. Use of the small molecule peptide of any one of claims 1-2, the fusion protein of claim 3 in the preparation of a product for improving butter cultivation efficiency.
  7. 7. A product for improving the cultivation efficiency of butter crabs, comprising the small molecule peptide according to any one of claims 1-2 and the fusion protein according to claim 3.
  8. 8. The product of claim 7, wherein: The product comprises pharmaceutically acceptable auxiliary materials.
  9. 9. A method for improving the cultivation efficiency of butter crabs is characterized by comprising the following steps: Administering the small molecule peptide of any one of claims 1-2, the fusion protein of claim 3, or the product of any one of claims 7-8 to a female blue crab.
  10. 10. The method for screening a small molecule peptide according to any one of claims 1 to 2, comprising the steps of: Screening the optimal binding conformation according to the predicted binding conformation of the vitellogenin and the receptor sequence thereof, screening hot spot residues according to the optimal binding conformation, and calculating the binding energy change before and after mutation of the hot spot residues to determine the competitive inhibition small molecule peptide sequence.

Description

Small molecular peptide for inhibiting development of blue crab ovary and application thereof Technical Field The invention belongs to the technical field of biology, and particularly relates to a small molecular peptide for inhibiting the development of blue crab ovary and application thereof. Background Scylla paramamosain (Scylla Pramamosain) is an important economic crab in southeast coastal China. The cultivation yield is kept to be more than or equal to 16 ten thousand tons and is kept to float continuously for several years, and the annual yield value is about 300 hundred million yuan. The novel vegetable basket has the advantages of enriching the vegetable basket for people and simultaneously providing a channel for improving the income of fishery practitioners, and has important roles in the aquaculture industry in China. The yellow crab is blue crab, which is a basic substance vitellogenin (Vitellogenin) generated by liver pancreas for ovary development can not enter ovary due to the abnormal development of ovary caused by various environmental factors, and is remained in blood, and the lipid in vitellogenin is degraded and flows through the whole body along with the blood circulation due to the change of environmental factors, so that the crab body is filled with golden yellow grease, and the joint membrane is golden and transparent, thus the yellow crab is called as yellow crab. The butter crab is used as a unique rare food material in the Yue-Kong-Australian Dawan in recent years, the price is kept at 1000 yuan/kg throughout the year, and the price of the mid-autumn festival can break through 2000 yuan/kg, which is 10 times of the market price of the ordinary blue crab. Butter crabs are found accidentally in traditional large-surface aquaculture ponds, and the occurrence probability is only one thousandth. Up to the present, a technical system for cultivating butter crabs in a large scale and production supporting facilities do not exist, so that the supply of butter crabs can not meet the consumption demands of the market, and the price of butter crabs is high. Therefore, the novel breed of butter crabs has huge consumer market and economic value. The cultivation mechanism of the butter crabs has attracted wide attention from scientific research institutions, cultivation enterprises and investors, but the current stable and high-yield cultivation technology system is still immature, and the research and development of various cultivation technologies and equipment are still under exploration. Disclosure of Invention Vitellogenin (vitellogenin Vtg) is a transferrin produced by scylla paramamosain by the liver and pancreas, transported to the ovary through blood and specifically recognized by vitellogenin receptor (Vitellogenin Receptor VtgR) into the ovary to promote its development, and is a basic substance for the development of the ovary of scylla paramamosain. The invention respectively acquires the protein sequences of the vitellogenin and the receptor thereof through NCBI database, intercepts 8 segments of sequences in the full-length sequence of the vitellogenin and 3 segments of sequences in the receptor through alkaline plaque scanning and protein domain analysis, and utilizes Alpha fold2 Multimer software to simulate and predict the binding mode of similar proteins in homologous species based on computer deep learning, wherein pLDDT, pTM, iPTM values of 24 pairs of sequence binding conformations are screened to obtain the optimal binding conformation. The potential hot spot residues in the optimal conformation, solvent and surface area in the binding interface (Δsasa) Top10, and residues that generate polar hydrogen bonds were then screened using Pymol software. And (3) taking the hot spot residue intersection, mutating the hot spot residues by using YASARA software, calculating delta G bind before and after mutation, and selecting the sequence of the hot spot residues +/-5-8 aa as the sequence of the competitive inhibition polypeptide according to delta G bind change. Through researches after swimming foot basal injection, the vitellogenin competitive inhibition polypeptide can obviously inhibit the development of blue crab ovary to induce the formation of yellow crabs. The small molecular peptide developed through computer aided learning inhibits the development of ovaries through competing binding sites of the vitellogenin and the vitellogenin receptor, and an auxiliary biological agent for improving the production efficiency of the butter crabs is constructed. The purpose of the invention is as follows: the invention aims at a small molecule peptide for inhibiting the development of blue crab ovary. The object of the second aspect of the present invention is to provide a fusion protein. The object of the third aspect of the present invention is to provide a biological material related to the small molecule peptide of the first aspect of the present invention and the fusion protein