Application of olfactory receptor in identification of 3-methylthio-propionaldehyde and method for detecting 3-methylthio-propionaldehyde

A technology of methylthiopropionaldehyde and olfactory receptors, which is applied to the use of olfactory receptors in recognizing 3-methylthiopropionaldehyde and the field of detection of 3-methylthiopropionaldehyde, which can solve the problem that 3-methylthiopropanal has not yet appeared. Propionaldehyde, etc.

Active Publication Date: 2022-07-08
HANVON CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there have been no reports on the olfactory receptors for the detection of 3-methylthiopropanal

Method used

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  • Application of olfactory receptor in identification of 3-methylthio-propionaldehyde and method for detecting 3-methylthio-propionaldehyde
  • Application of olfactory receptor in identification of 3-methylthio-propionaldehyde and method for detecting 3-methylthio-propionaldehyde
  • Application of olfactory receptor in identification of 3-methylthio-propionaldehyde and method for detecting 3-methylthio-propionaldehyde

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Example 1: Screening of olfactory receptors

[0078] In this example, the dual-luciferase method (Dual-Glo™ Luciferase Assay System, Promega) was used to measure the activity of olfactory receptors. The inventors selected some olfactory receptors from human and mouse olfactory receptor libraries, then prepared gene constructs containing olfactory receptors, Golf, CRE-Luciferase and pRL-SV40, and used the transfection reagent Lipofectamine2000 (Invitrogen) to transfer the olfactory receptors. The gene constructs were transfected into HEK293T cells. After culturing for 24 hours, 3-methylthiopropanal was diluted with culture medium to a concentration of 500 μM, cells were stimulated with the diluted 3-methylthiopropanal, and incubated for 2-4 hours. If the olfactory receptor is activated, the intracellular cAMP concentration increases, cAMP will bind to the promoter region of CRE-luciferase, and promote the transcription and translation of luciferase. By detecting the act...

Embodiment 2

[0088] Example 2: Response of four olfactory receptors to different compounds

[0089] In this example, the method of Example 1 was used to detect the responses of MOR244-3, MOR256-17, MOR180-1 and OR2C1 to three structural analogs of 3-methylthiopropanal (also known as "odor molecules"), The three structural analogs of 3-methylthiopropanal (methional) are 3-methylthiopropanol (methionol), n-pentanal (pentanal) and pentanol (pentanol), the specific test results are shown in Table 1 . Among them, the hydroxyl group in 3-methylthiopropanol replaces the aldehyde group of 3-methylthiopropanal; the carbon atom in n-pentanal replaces the sulfur atom of 3-methylthiopropanal; the hydroxyl and carbon atoms in pentanol At the same time, the aldehyde group and sulfur atom of 3-methylthiopropanal were replaced. In the table, "+++" means the response change fold ≥ 10, "++" means the response change fold 5

Embodiment 3

[0092] Example 3: Detection of 3-methylthiopropanal in different freshly squeezed fruit juices by MOR244-3 and MOR256-17

[0093] For the detection method in this example, refer to Example 1. The only difference is that in this example, a luciferase variant is used to replace the original dual luciferase, and the olfactory receptors expressing MOR244-3 and MOR256-17 are used respectively. The presence of 3-methylthiopropanal in freshly squeezed orange juice and freshly squeezed mango juice was detected by the cells of the body. For the specific test results, see figure 2 . The results show a peak around 15 minutes (min) with strong signals from both olfactory receptors.

[0094] Cells were pre-expressed with a luciferase variant by co-transfection of the pGloSensor-20F cAMP (Promega) construct. After activation of olfactory receptors, an increase in cAMP concentration can cause a conformational change of luciferase variants, which converts luciferase from an inactive state ...

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Abstract

The invention relates to application of an olfactory receptor in identification of 3-methylthiopropionaldehyde and a method for detecting the 3-methylthiopropionaldehyde. The invention relates to an application of an olfactory receptor in identification of 3-methylthiopropionaldehyde. The olfactory receptor comprises at least one selected from the group consisting of MOR244-3, MOR256-17, MOR180-1 and OR2C1, and the 3-methylthiopropionaldehyde is selected from the group consisting of MOR244-3, MOR256-17, MOR180-1 and OR2C1. The method comprises the steps that a to-be-detected sample makes contact with an olfactory receptor, the response value of the olfactory receptor is determined, whether the to-be-detected sample contains 3-methylthiopropionaldehyde or not is determined based on the response value, and the olfactory receptor comprises at least one selected from MOR244-3, MOR256-17, MOR180-1 and OR2C1. The olfaction receptor disclosed by the invention can be used for rapidly identifying the 3-methylthio-propionaldehyde and can be used for detecting the 3-methylthio-propionaldehyde in drinks, fruits or fruit products and other products so as to determine the freshness degree of the products or the types of the fruits contained in the products.

Description

technical field [0001] The present invention relates to the technical field of chemical detection, in particular, the present invention relates to the use of an olfactory receptor in recognizing 3-methylthiopropanal and a method for detecting 3-methylthiopropanal, more particularly, the present invention relates to Use of an olfactory receptor in recognizing 3-methylthiopropanal, use of 3-methylthiopropanal in activating olfactory receptors, method for detecting 3-methylthiopropanal, detection of wine, fruit or Methods for the freshness of fruit products, methods for detecting the type of fruit contained in food, methods for distinguishing between beer or red wine, and methods for monitoring whether fruit products have undergone high-temperature sterilization treatment. Background technique [0002] Olfactory receptors (ORs), also known as odorant receptors, are chemoreceptors expressed in the cell membrane of olfactory neurons (OSNs). Olfactory receptors activated by odora...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/02G01N33/14
CPCG01N33/02G01N33/025G01N33/146
Inventor 刘卫红
Owner HANVON CORP
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