Application of organic selenium compound ASea-23 in preparation of selenium-enriched wheat

Abstract

The invention relates to an application of an organic selenium compound ASea-23 in preparation of selenium-enriched wheat. According to the invention, a small molecule ASea-23 which is highly compatible with a wheat selenium absorption key factor-ATP (adenosine triphosphate) sulfurase is screened out; the ASea-23 is independently used or mixed with other components to prepare liquid, solid or aerosol, and the liquid, solid or aerosol is used for wheat in the modes of foliage spraying, soil application or seed soaking and the like, so that the ASea-23 maintains a certain concentration in wheat plants, the selenium absorption efficiency of the wheat can be greatly promoted, the selenium-rich wheat is further prepared, and the selenium-rich wheat is obtained. The technological innovation has a good industrial application prospect.

Description

Technical Field

[0001] The invention relates to the field of grain cultivation, and in particular to application of an organic selenium compound ASea-23 in the preparation of selenium-enriched wheat. Background Art

[0002] Selenium is an essential trace element for the human body and has many positive effects on human health. For example, selenium can protect cells from free radical damage, has anti-aging effects, improves human immunity, protects the health of the heart and blood vessels, prevents Keshan disease and cardiomyopathy, and antagonizes harmful heavy metals. However, the human body cannot synthesize selenium on its own, and dietary selenium is the main source for humans. In view of this, we need to increase the content of selenium in the diet. Wheat is the first staple food for humans at this stage, and producing selenium-enriched wheat is an important way to increase the selenium content in the diet. However, due to the scarcity of selenium in the soil and the limited selenium absorption capacity of wheat, it is difficult and costly to produce selenium-enriched wheat. Therefore, we are required to develop a selenium-containing compound that can be highly efficient for wheat. Summary of the invention

[0003] In view of the defects in the prior art, the object of the present invention is to provide an application of an organic selenium compound ASea-23 in the preparation of selenium-enriched wheat.

[0004] In order to achieve the above purpose, the technical solution adopted by the present invention is:

[0005] An application of an organic selenium compound ASea-23 in preparing selenium-enriched wheat, characterized in that the Asea-23 is acted on wheat to cultivate selenium-enriched wheat;

[0006] The structural formula of ASea-23 is:

[0007]

[0008] The method of applying Asea-23 to wheat includes: applying ASea-23 alone or in the form of a biological agent to wheat by foliar spraying, soil application or seed soaking. The dosage form of the biological agent includes liquid, solid or aerosol.

[0009] A biological preparation, characterized in that the biological preparation contains Asea-23, and the dosage form of the biological preparation includes liquid, solid or aerosol.

[0010] ASea-23 referred to in the present invention is a small molecule compound containing selenium, and its standard name is:

[0011] (4-Phenylselanylphenyl)5,6-bis(4-hydroxyphenyl)-7-oxabi cyclo[2.2.1]hept-5-ene-2-sulfonate, PubChemCID is 168279125;

[0012] Its Smiles style is

[0013] C1C(C2C(=C(C1O2)C3=CC=C(C=C3)O)C4=CC=C(C=C4)O)S(=O)(=O)OC5=CC=C(C=C5)[Se]C6=CC=CC=C6,

[0014] The relative molecular mass of ASea-23 is 591.5, and the structural formula is as follows: Figure 1 shown.

[0015] The ATP sulfurylase referred to in the present invention is also known as ATP sulfurylase, etc. The most basic function is to transfer sulfate to the adenine monophosphate group of ATP. Since selenium and sulfur share a common metabolic pathway in plants, the absorption of selenate by plants is mediated by sulfur transporters. Many studies have shown that plants overexpressing the ATP sulfurylase gene show 3-5 times enhanced selenium accumulation characteristics regardless of experimental control conditions or field high-selenium soil conditions. Many studies by this research group have also found that the enzyme may be the rate-limiting enzyme in plant selenate metabolism. Finding small molecule selenium compounds with strong affinity to the above enzyme has greatly promoted the selenium absorption effect of wheat.

[0016] The application of the organic selenium compound ASea-23 in the preparation of selenium-enriched wheat has the following beneficial effects:

[0017] By utilizing the strong affinity of the organic selenium compound ASea-23 with ATP sulfurylase, wheat can achieve good absorption of selenium at a lower concentration. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The present invention has the following accompanying drawings:

[0019] Figure 1 is the structural formula of the organoselenium compound ASea-23 of the present invention;

[0020] Figure 2 Schematic diagram of the binding site of ASea-23 and ATP sulfurylase docking product;

[0021] Figure 3 The calculation results of the interaction force and chemical bond between ASea-23 and ATP sulfurylase;

[0022] Figure 4 The stability results of the complex between ASea-23 and ATP sulfurylase were simulated by AMBER molecular dynamics.

[0023] Figure 5 This is a quantitative analysis diagram of the interaction between ASea-23, ATP sulfurylase complex and protein residues. DETAILED DESCRIPTION

[0024] The present invention is further described in detail below in conjunction with the accompanying drawings.

[0025] The inventors used AlphaFold2 to study the three-dimensional structure of wheat ATP sulfurylase and found pockets where small molecules can enter and stably stay. Based on selenium-containing small molecule compounds in small molecule databases such as PubChem, they used AutoDockVina software for high-throughput docking, searched and annotated the 500 small molecules with the highest binding free energy (ΔGbinding) in the output results, and used the Internet to search their physical, chemical and physiological properties. After comprehensive consideration, they found that the physiological properties of 113 compounds such as ASea-23 are most suitable for the preparation of selenium-enriched wheat. The inventors used AutoDock Vina software to perform fine docking on these 113 compounds. The binding free energy ranking obtained from the fine docking, for example, the binding free energy of ASea-23 is -9.6kcal / mol, and the corresponding half-effective concentration (EC50) is calculated to be 86nM using the formula: EC50≈exp(ΔGbinding / RT). The binding site of the docking product is as follows: Figure 2 The interaction force and chemical bond between ASea-23 and ATP sulfurylase were calculated using Protein-Ligand Interaction Profiler on the website "https: / / projects.biotec.tu-dresden.de / plip-web / plip". The results are as follows Figure 3 As shown, molecular dynamics simulations were performed using AM BER and other methods to determine the stability of the complex ( Figure 4 ) and quantitative analysis of interactions with protein residues ( Figure 5 ), and found that the complex had good stability and the interaction simulation was consistent with expectations.

[0026] A high-throughput standardized experiment was established to evaluate the actual effectiveness of several affinity agents screened above:

[0027] The wheat ATP sulfurylase sequence was downloaded from the NCBI database, and its full-length primers were designed using Snapgene and submitted to Bio-Tech for synthesis. The root samples of common wheat were separated and cleaned, and RNA was extracted. The obtained RNA was reverse transcribed into cDNA, and the target gene was amplified and purified. After gel recovery, the target gene and the vector were connected by homologous recombination. The ligation product was transformed into competent Escherichia coli, and the positive clones were screened and identified. After the plasmid was extracted, it was transformed and amplified again, and the ATP sulfurylase was prepared and purified. After the concentration and amount met the requirements, the SPR-based Biacore instrument was used to detect the binding affinity of ATP sulfurylase and small molecule compounds.

[0028] Referring to the binding free energy of each derivative determined in the previous screening work, the corresponding measured EC50 was calculated respectively, specifically:

[0029] The EC50 values ​​of each derivative were predicted by taking the multiple ratio gradient, and the measured EC50 values ​​of each substance were detected by using the isolated and purified ATP sulfurylase. The experimental results showed that ASea-23 had the best affinity for ATP sulfurylase, with a measured EC50 of 0.17 μM.

[0030] The contents not described in detail in this specification belong to the prior art known to professional and technical personnel in this field.

Claims

1. An application of an organic selenium compound ASea-23 in the preparation of selenium-enriched wheat, characterized in that: The Asea-23 is applied to wheat to cultivate selenium-enriched wheat; The structural formula of ASea-23 is: The methods of applying Asea-23 to wheat include: applying ASea-23 alone or as a biological agent to wheat by foliar spraying, soil application or seed soaking.

2. A biological agent, characterized in that: The biological preparation contains Asea-23, and the dosage form of the biological preparation includes liquid, solid or aerosol.

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