CN-121974993-A - Altai hepialus antifreeze protein and application thereof

Abstract

The invention discloses an Altai hepialus antifreeze protein and application thereof, and relates to the technical field of antifreeze. The amino acid sequence of the antifreeze protein is shown as SEQ ID NO. 1. The amino acid sequence of the Altai Hepialus antifreeze protein takes 76 amino acids as a unit, circulates 7 times, mainly takes MQIFVKTLTGKTITLEVE as a main component, and has no homology with the antifreeze protein of the prior reported insects.

Inventors

• WANG YAN
• YIN YIQUN
• TAN TINGTING
• XU XIN
• LIU XINHUI

Assignees

• 石河子大学

Dates

Publication Date

20260505

Application Date

20260126

Claims (4)

1. 1. The Altai hepialus antifreeze protein is characterized in that the antifreeze protein has an amino acid sequence shown as SEQ ID NO. 1.
2. 2. The anti-freeze protein of Altai Hepialus according to claim 1, wherein the amino acid sequence shown in SEQ ID NO. 1 is replaced, deleted or inserted with one or more amino acid residues to form a protein molecule with anti-freeze function.
3. 3. A gene sequence encoding the alpialus antifreeze protein of claim 1, wherein said gene sequence is (a) or (b): (a) The nucleotide sequence is shown as SEQ ID NO. 2; (b) The nucleotide sequence shown in SEQ ID No.2 is replaced, deleted or inserted with one or several nucleotides to form the gene sequence encoding the Altai Hepialus antifreeze protein as defined in claim 1.
4. 4. Use of the anti-freeze protein of Altai Hepialus in the anti-freeze field.

Description

Altai hepialus antifreeze protein and application thereof Technical Field The invention relates to the technical field of freezing resistance, in particular to an Altai hepialus freezing resistance protein and application thereof. Background Antifreeze proteins (Antifreeze Proteins, AFPs), also known as ice binding proteins (Ice Binding Proteins, IBPs), are a special protein that is formed and expressed during long-term evolution of cold-region organisms to adapt to the severe cold environment. AFPs are capable of binding to a specific site of ice crystals, and by inhibiting the growth rate thereof on that specific site, without altering the growth rate at other sites, thereby altering the normal spherical growth morphology of ice crystals, exhibiting a unique change in ice crystal morphology, known as ice crystal modification activity. Ice crystal modification activity is the most easily observed activity of antifreeze proteins and is the classical way to test for the presence or absence of antifreeze proteins. Furthermore, AFPs are able to non-quantitatively lower the freezing point of a solution without significantly affecting its melting point, thereby creating a difference between the freezing and melting points, a property known as thermal hysteresis (THERMAL HYSTERESIS ACTIVITY, THA), and therefore this protein is also known as thermal hysteresis protein (THERMAL HYSTERESIS Proteins, THPs). According to the source, AFPs are mainly classified into fish-derived antifreeze proteins, plant-derived antifreeze proteins, bacterial-derived antifreeze proteins and insect-derived antifreeze proteins. Notably, insect-derived antifreeze proteins generally exhibit higher antifreeze activity than other sources, and are therefore of greater interest to researchers. More than twenty insects with AFPs have been reported so far, and more studies at home and abroad mainly include yellow meal worm (Tenebrio molitor), red wing beetle (Dendroides canadensis), spruce leaf roller (Choristoneura fumiferana), xinjiang desert beetles (Microdera punctipennis) and (Anatolica polita) and the like. Most of them belong to the order coleoptera, while there is relatively little research into lepidopteran AFPs. Therefore, the provision of a novel Altai hepialus antifreeze protein and its use are a problem to be solved by those skilled in the art. Disclosure of Invention In view of this, the invention provides an Altai Hepialus antifreeze protein and application thereof. In order to achieve the above purpose, the present invention adopts the following technical scheme: an Altai hepialus antifreeze protein, the antifreeze protein number is Gene.73409, and the amino acid sequence is shown as SEQ ID NO. 1. Furthermore, the protein molecule with the antifreeze function is formed by replacing, deleting or inserting one or more amino acid residues in the amino acid sequence shown as SEQ ID NO. 1. A gene sequence encoding said alcian hepialus antifreeze protein, said gene sequence being (a) or (b): (a) The nucleotide sequence is shown as SEQ ID NO. 2; (b) The nucleotide sequence shown in SEQ ID NO.2 is replaced, deleted or inserted with one or more nucleotides to form a gene sequence for encoding the Altai Hepialus antifreeze protein. Application of Altai hepialus antifreeze protein in the antifreeze field. Compared with the prior art, the invention has the beneficial effects that: The invention takes frozen but dead lepidoptera insect, namely Altai hepialus (Hepialus altaicola), as a research object, and samples are collected in Altaishan mountain with high latitude and high altitude in the Uygur autonomous region of Xinjiang. Altai hepialus is the host insect of Cordyceps sinensis. The research shows that the larvae can still survive after in vivo icing, and the larvae show extremely strong freezing resistance. Therefore, it is presumed that the anti-freezing protein resisting freezing may exist in the Altai hepialus body, and the early experiments prove that the haemolymph has obvious ice crystal modification activity. In 2021, the inventor topic group has identified an Altai hepialus antifreeze protein sequence and obtained the patent of the related invention. However, according to the previous studies of the inventors, it is presumed that such an antifreeze protein may be present in the body. However, when the overwintering Albata is subjected to transcriptome sequencing previously, in the transcriptome data thereof, a gene sequence newly annotated as an antifreeze protein is not obtained except for the antifreeze protein previously found by the inventor, which indicates that a novel antifreeze protein which has not been reported yet has yet been discovered. Therefore, the inventor develops a new set of natural antifreeze protein extraction method, separates a special protein with ice crystal modification activity from Altai Hepialus, and obtains the peptide sequence through chromatographic purification and mass spectrum sequencing. The