CN115840026A - Use of olfactory receptor for recognizing 4-methoxybenzaldehyde and method for detecting 4-methoxybenzaldehyde - Google Patents
Use of olfactory receptor for recognizing 4-methoxybenzaldehyde and method for detecting 4-methoxybenzaldehyde Download PDFInfo
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Abstract
The invention provides a method for detecting 4-methoxybenzaldehyde. The method comprises the following steps: contacting a sample to be detected with an olfactory receptor, and determining a response value of the olfactory receptor after the contact; determining whether the sample to be detected contains 4-methoxybenzaldehyde or not based on the response value; wherein the olfactory receptor comprises OR2J2 and/OR OR2W1. The 4-methoxybenzaldehyde in the method can activate the olfactory receptor, so that a sample to be detected is contacted with the olfactory receptor, if the sample to be detected contains the 4-methoxybenzaldehyde, the olfactory receptor can be activated, a response value after the olfactory receptor is activated is obtained, and whether the sample to be detected contains the 4-methoxybenzaldehyde or not can be determined according to the response value.
Description
Technical Field
The invention relates to the technical field of chemical detection, in particular to application of an olfactory receptor in identifying 4-methoxybenzaldehyde and a method for detecting 4-methoxybenzaldehyde, and more particularly relates to application of the olfactory receptor in identifying 4-methoxybenzaldehyde, application of 4-methoxybenzaldehyde in activating the olfactory receptor, a method for detecting 4-methoxybenzaldehyde, a method for detecting food raw materials and a method for evaluating quality of perfume products.
Background
The 4-methoxybenzaldehyde (CAS number 123-11-5), commonly known as anisaldehyde (p-anisaldehyde), has a molecular structure comprising a benzene ring, a methoxy group and an aldehyde group, and has strong anise and hawthorn fragrance. Natural products are found in fennel, star anise, vanilla beans, acacia, and the like.
The 4-methoxy benzaldehyde has the following applications: 1) The method is mainly used for preparing daily and food essences such as vanilla, spice, apricot, cream, fennel, caramel, cherry, chocolate, walnut, raspberry, strawberry, mint and the like; 2) 4-methoxy benzaldehyde is a good brightener which is specified as edible spice temporarily allowed to be used in China GB2760-86 and used as a cyanide-free galvanizing DE additive, can improve anode polarization in a wider current range, obtains a bright coating and creates favorable conditions for environmental protection; 3) In the pharmaceutical industry are intermediates for antihistamine drugs for the manufacture of the antimicrobial drug hydroxyaminobenzylpenicillin; 4) Compounds with functional groups of nitrogen atoms were followed as developers.
Human beings have excellent olfactory perception ability and can sensitively and rapidly recognize and distinguish odors in daily foods, mainly because odorants can activate human Olfactory Receptors (ORs) to trigger nerve impulses, transmitting information about odors to the brain. About 400 olfactory receptors are known in humans, but no report has been made on detection of olfactory receptors in 4-methoxybenzaldehyde.
Disclosure of Invention
The present invention aims to solve at least to some extent at least one of the technical problems of the prior art. Therefore, the invention provides the use of OR2J2 and/OR OR2W1 olfactory receptor for identifying 4-methoxybenzaldehyde, the olfactory receptor can rapidly identify 4-methoxybenzaldehyde, can detect 4-methoxybenzaldehyde in perfume products, and can be used for evaluating the quality of the perfume products.
In a first aspect of the invention, the invention proposes the use of an olfactory receptor comprising OR2J2 and/OR 2W1 for the recognition of 4-methoxybenzaldehyde. The inventor experimentally found that after the stimulation of OR2J2 and OR2W1 with 4-methoxybenzaldehyde, respectively, OR2J2 and OR2W1 can be activated, so that the olfactory receptor can be used for effectively identifying 4-methoxybenzaldehyde, and a foundation is laid for the identification of 4-methoxybenzaldehyde and the detection of samples containing 4-methoxybenzaldehyde.
In a second aspect of the invention, the invention proposes the use of 4-methoxybenzaldehyde for activating olfactory receptors comprising OR2J2 and/OR 2W1. The inventor experimentally found that after the stimulation of OR2J2 and OR2W1 with 4-methoxybenzaldehyde, respectively, OR2J2 and OR2W1 can be activated, so that the olfactory receptor can be used for effectively identifying 4-methoxybenzaldehyde, and a foundation is laid for the identification of 4-methoxybenzaldehyde and the detection of samples containing 4-methoxybenzaldehyde.
In a third aspect of the present invention, the present invention provides a method for detecting 4-methoxybenzaldehyde. According to an embodiment of the invention, the method comprises: contacting a sample to be detected with an olfactory receptor, and determining a response value of the olfactory receptor after the contact; determining whether the sample to be detected contains 4-methoxybenzaldehyde or not based on the response value; wherein the olfactory receptor comprises OR2J2 and/OR OR2W1. It can be known from the foregoing that 4-methoxybenzaldehyde can activate the olfactory receptor, so that a sample to be tested is contacted with the olfactory receptor, and if the sample to be tested contains 4-methoxybenzaldehyde, the olfactory receptor can be activated, and a response value after the olfactory receptor is activated is obtained, and whether the sample to be tested contains 4-methoxybenzaldehyde can be determined according to the response value.
In a fourth aspect of the invention, a method of detecting a food material is presented. According to an embodiment of the invention, the method comprises: contacting a sample to be detected with an olfactory receptor, and determining a response value of the olfactory receptor after contact; determining the raw material of the sample to be detected based on the response value; wherein the olfactory receptor comprises OR2J2 and/OR OR2W1. It can be known from the foregoing that 4-methoxybenzaldehyde can activate the olfactory receptor, so that a sample to be tested is brought into contact with the olfactory receptor, and if the sample to be tested contains 4-methoxybenzaldehyde, the olfactory receptor can be activated, and a response value after the olfactory receptor is activated is obtained, it can be determined that a raw material containing 4-methoxybenzaldehyde exists in the sample to be tested, and if the olfactory receptor does not have a response value, the raw material containing 4-methoxybenzaldehyde does not exist in the sample to be tested.
In a fifth aspect of the invention, a method of assessing the quality of a fragrance product is presented. According to an embodiment of the invention, the method comprises: contacting a perfume product to be detected with an olfactory receptor, and determining a response value of the olfactory receptor after the contact; determining the quality of the perfume product to be tested based on the response value; wherein the olfactory receptor comprises OR2J2 and/OR OR2W1. It can be known from the foregoing that 4-methoxybenzaldehyde can activate the olfactory receptor, so that the perfume product to be tested is contacted with the olfactory receptor, and if the perfume product to be tested contains 4-methoxybenzaldehyde, the olfactory receptor can be activated, and a response value after the olfactory receptor is activated is obtained, and the quality of the perfume product to be tested can be determined according to the magnitude of the response value.
Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.
Drawings
The above and/or additional aspects and advantages of the present invention will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:
FIG. 1 is a graph of the responses of 4-methoxybenzaldehyde to the stimulation of 398 olfactory receptors (hOR) in human in example 1 of the present invention;
FIG. 2 is a dose-dependent curve of OR2J2 and OR2W1 responses to 4-methoxybenzaldehyde in example 1 of the present invention;
FIG. 3 is a dose-dependent curve of OR2J2 response to the affluent brand anise and fennel extract of example 2 of the present invention;
FIG. 4 is a graph showing the dose-dependence of OR2J2 response to the self-produced star anise oil and plant Yisheng brand star anise oil of Guangxi farmer in example 3 of the present invention;
FIG. 5 shows the response of OR2J2 to the remitted brand fennel gas in example 4 of the present invention.
Detailed Description
The following describes embodiments of the present invention in detail. The following examples are illustrative only and are not to be construed as limiting the invention.
It should be noted that the terms "first" and "second" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. Further, in the description of the present invention, "a plurality" means two or more unless otherwise specified.
The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value, and such ranges or values should be understood to encompass values close to those ranges or values. For ranges of values, between the endpoints of each of the ranges and the individual points, and between the individual points may be combined with each other to give one or more new ranges of values, and these ranges of values should be considered as specifically disclosed herein.
In order that the invention may be more readily understood, certain technical and scientific terms are specifically defined below. Unless clearly defined otherwise herein, all other technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
In this document, the terms "comprise" or "comprise" are open-ended expressions that include the elements indicated in the present invention, but do not exclude other elements.
As used herein, the terms "optionally," "optional," or "optionally" generally mean that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where the event or circumstance occurs, and instances where it does not.
The present invention provides a use of an olfactory receptor for recognizing 4-methoxybenzaldehyde, a use of 4-methoxybenzaldehyde for activating the olfactory receptor, a method for detecting 4-methoxybenzaldehyde, a method for detecting a food material, and a method for evaluating the quality of a flavor product, which will be described in detail, respectively, below.
Use of
In a first aspect of the invention, the invention proposes the use of an olfactory receptor comprising OR2J2 and/OR 2W1 for the recognition of 4-methoxybenzaldehyde. The inventor finds out through experiments that after the stimulation of OR2J2 and OR2W1 by 4-methoxybenzaldehyde, OR2J2 and OR2W1 can be activated, so that the olfactory receptor can be used for effectively identifying the 4-methoxybenzaldehyde, and the method lays a foundation for the identification of the 4-methoxybenzaldehyde and the detection of samples containing the 4-methoxybenzaldehyde. The inventor further finds that the olfactory receptor can recognize 4-methoxybenzaldehyde gas in the same space, and provides possibility for bionic olfactory recognition.
It should be noted that if 4-methoxybenzaldehyde stimulates the olfactory receptor, the olfactory receptor is activated, i.e. "the olfactory receptor can recognize 4-methoxybenzaldehyde"; if 4-methoxybenzaldehyde stimulates olfactory receptor, the olfactory receptor is not activated, namely, the olfactory receptor can not recognize 4-methoxybenzaldehyde.
In a preferred embodiment of the invention, the olfactory receptor is OR2J2.
According to an embodiment of the invention, said identification is represented by a change in the activity of an olfactory receptor.
According to an embodiment of the invention, the activity change comprises at least one of the following signal changes: luciferase, secreted alkaline phosphatase, fluorescent protein, fluorescent probe, cAMP, IP3, calcium ion, current, and pH.
Illustratively, the recognition is manifested by an increase in cAMP downstream of the olfactory receptor. The inventor finds out through experiments that 4-methoxybenzaldehyde stimulates cells expressing olfactory receptors, the cAMP concentration in the cells is increased after the olfactory receptors are activated, and whether the olfactory receptors can recognize the 4-methoxybenzaldehyde can be determined by detecting the change of the cAMP concentration.
In a second aspect of the invention, the invention proposes the use of 4-methoxybenzaldehyde in the activation of olfactory receptors comprising OR2J2 and/OR 2W1. The inventor experimentally found that after the stimulation of OR2J2 and OR2W1 with 4-methoxybenzaldehyde, respectively, OR2J2 and OR2W1 can be activated, so that the olfactory receptor can be used for effectively identifying 4-methoxybenzaldehyde, and a foundation is laid for the identification of 4-methoxybenzaldehyde and the detection of samples containing 4-methoxybenzaldehyde.
In a preferred embodiment of the invention, the olfactory receptor is OR2J2.
According to an embodiment of the invention, said activation is represented by a change in the activity of olfactory receptors.
According to an embodiment of the invention, said activity change comprises at least one of the following signal changes: luciferase, secreted alkaline phosphatase, fluorescent protein, fluorescent probe, cAMP, IP3, calcium ion, current, and pH.
Illustratively, the activation is manifested by an increase in cAMP downstream of the olfactory receptor. The inventor finds out through experiments that 4-methoxybenzaldehyde stimulates cells expressing olfactory receptors, the cAMP concentration in the cells is increased after the olfactory receptors are activated, and whether the olfactory receptors can activate the 4-methoxybenzaldehyde can be determined by detecting the change of the cAMP concentration.
Method
In a third aspect of the present invention, the present invention provides a method for detecting 4-methoxybenzaldehyde. According to an embodiment of the invention, the method comprises: contacting a sample to be detected with an olfactory receptor, and determining a response value of the olfactory receptor after the contact; determining whether the sample to be detected contains 4-methoxybenzaldehyde or not based on the response value; wherein the olfactory receptor comprises OR2J2 and/OR OR2W1. It can be known from the foregoing that 4-methoxybenzaldehyde can activate the olfactory receptor, so that a sample to be tested is contacted with the olfactory receptor, and if the sample to be tested contains 4-methoxybenzaldehyde, the olfactory receptor can be activated, and a response value after the olfactory receptor is activated is obtained, and whether the sample to be tested contains 4-methoxybenzaldehyde can be determined according to the response value.
It should be noted that "contacting" in this document should be understood broadly, and may be a direct contact or an indirect contact, and is not limited specifically.
Illustratively, a sample to be tested (which is liquid or prepared into liquid by using a solvent) in a liquid state is mixed with an olfactory receptor and then contacted (namely, the sample to be tested and the olfactory receptor are directly contacted); or the sample to be detected and the olfactory receptor can be directly placed in the same space, and the odor molecules (4-methoxybenzaldehyde) released by the sample to be detected are contacted with the olfactory receptor (namely, the sample to be detected and the olfactory receptor are indirectly contacted).
In a preferred embodiment of the invention, the olfactory receptor is OR2J2.
According to an embodiment of the present invention, said olfactory receptor present response value is an indication that 4-methoxybenzaldehyde is contained in said sample to be tested; alternatively, the absence of a response value by the olfactory receptor is indicative of the absence of 4-methoxybenzaldehyde in the test sample.
It should be noted that the "indication that 4-methoxybenzaldehyde is not contained" means that 4-methoxybenzaldehyde is completely absent from the sample to be tested; or a small amount of 4-methoxybenzaldehyde is present in the sample to be tested but cannot be detected.
According to an embodiment of the present invention, said olfactory receptor is selected from the group consisting of OR2J2, and said olfactory receptor presence response value is an indication that 4-methoxybenzaldehyde is contained OR an indication that 4-methoxybenzaldehyde is contained at not less than 30 μ M in said sample to be tested; alternatively, the absence of response value by the olfactory receptor is indicative of no 4-methoxybenzaldehyde or less than 30 μ M4-methoxybenzaldehyde in the sample.
According to an embodiment of the present invention, said olfactory receptor is selected from the group consisting of OR2W1, and said olfactory receptor presence response value is an indication that 4-methoxybenzaldehyde is contained OR an indication that 4-methoxybenzaldehyde is contained at a concentration of not less than 100 μ M in said sample to be tested; alternatively, the absence of response value by the olfactory receptor is indicative of no 4-methoxybenzaldehyde or less than 100 μ M4-methoxybenzaldehyde in the sample being tested.
According to an embodiment of the invention, the method further comprises: and determining the content of the 4-methoxybenzaldehyde in the sample to be detected based on a standard curve, wherein the standard curve is a curve corresponding to the predetermined amount of the 4-methoxybenzaldehyde and the olfactory receptor response value. Therefore, the content of the 4-methoxybenzaldehyde in the sample to be detected can be detected.
According to an embodiment of the invention, the olfactory receptor is provided by a cell or transgenic cell expressing the olfactory receptor.
According to an embodiment of the invention, the cell or transgenic cell is a eukaryotic cell or a prokaryotic cell.
In some alternative embodiments of the invention, the eukaryotic cells include, but are not limited to, cells selected from the group of cells isolated from the olfactory substrate, HEK293 cells, CHO cells, xenopus oocytes, hela cells, COS cells, yeast cells, and the like.
According to an embodiment of the invention, the prokaryotic cell is selected from bacteria.
According to an embodiment of the invention, said response value is obtained by detecting a change in activity of said olfactory receptor.
According to an embodiment of the invention, said activity change is determined by at least one of the following detection methods: luciferase assay, secreted alkaline phosphatase assay, fluorescent protein assay, fluorescent probe assay, ca 2+ Concentration detection, current detection, isotopic labeling, antibody detection, and pH detection.
Illustratively, the luciferase assay is a method in which when cells containing olfactory receptors are stimulated with 4-methoxybenzaldehyde or a sample containing 4-methoxybenzaldehyde, if the olfactory receptors are activated, the intracellular concentration of cAMP increases, cAMP binds to the promoter region of CRE-luciferase and promotes transcription and translation of luciferase, and thus, by detecting the activity of luciferase, the response of the olfactory receptors can be characterized, and it can be determined whether the olfactory receptors recognize 4-methoxybenzaldehyde.
According to an embodiment of the invention, the response value is obtained by detecting a change in cAMP concentration in the cell.
Illustratively, the change in cAMP concentration in the cells is obtained by using the glosensort (tm) cAMP detection kit. The GloSensor-20F cAMP gene construct can pre-express a luciferase variant, the increase of cAMP concentration can cause the conformational change of the luciferase variant, so that the luciferase is converted from an inactive state to an active state, and the GloSensor (TM) cAMP detection kit provides a substrate of the luciferase variant.
In a fourth aspect of the invention, a method of detecting a food material is presented. According to an embodiment of the invention, the method comprises: contacting a sample to be detected with an olfactory receptor, and determining a response value of the olfactory receptor after contact; determining the raw material of the sample to be detected based on the response value; wherein the olfactory receptor comprises OR2J2 and/OR OR2W1. It can be known from the foregoing that 4-methoxybenzaldehyde can activate the olfactory receptor, so that a sample to be tested is brought into contact with the olfactory receptor, and if the sample to be tested contains 4-methoxybenzaldehyde, the olfactory receptor can be activated, and a response value after the olfactory receptor is activated is obtained, it can be determined that a raw material containing 4-methoxybenzaldehyde exists in the sample to be tested, and if the olfactory receptor does not have a response value, the raw material containing 4-methoxybenzaldehyde does not exist in the sample to be tested.
In a preferred embodiment of the invention, the olfactory receptor is OR2J2.
According to an embodiment of the present invention, the olfactory receptor presence response value is an indication that at least one of aniseed, fennel, vanilla beans, dill, corn and acacia is contained in the sample to be tested; or, the absence of a response value by the olfactory receptor is an indication that the sample to be tested does not contain anise, fennel, vanilla beans, dill, corn, and acacia.
According to the embodiment of the invention, the mass-to-volume ratio of the perfume raw materials in the sample to be tested is not lower than 0.01g/ml, such as not lower than 0.02g/ml, 0.03 g/ml, 0.04g/ml, 0.05 g/ml, 0.06g/ml, 0.07 g/ml, 0.08g/ml and the like.
According to an embodiment of the invention, the olfactory receptor is provided by a cell or transgenic cell expressing the olfactory receptor.
According to an embodiment of the invention, the cell or transgenic cell is a eukaryotic cell or a prokaryotic cell.
According to embodiments of the present invention, the eukaryotic cells include, but are not limited to, cells selected from the group of cells isolated from olfactory substrate, HEK293 cells, CHO cells, xenopus oocytes, hela cells, COS cells, yeast cells, and the like.
According to an embodiment of the invention, the prokaryotic cell is selected from bacteria.
According to an embodiment of the invention, said response value is obtained by detecting a change in activity of said olfactory receptor.
According to an embodiment of the invention, said activity change is determined by at least one of the following detection methods: luciferase assay, secreted alkaline phosphatase assay, fluorescent protein assay, fluorescent probe assay, ca 2+ Concentration detection, current detection, isotopic labeling, antibody detection, and pH detection.
Illustratively, the luciferase assay is a method in which when cells containing olfactory receptors are stimulated with 4-methoxybenzaldehyde or a sample containing 4-methoxybenzaldehyde, if the olfactory receptors are activated, the intracellular concentration of cAMP increases, cAMP binds to the promoter region of CRE-luciferase and promotes transcription and translation of luciferase, and thus, by detecting the activity of luciferase, the response of the olfactory receptors can be characterized, and it can be determined whether the olfactory receptors recognize 4-methoxybenzaldehyde.
According to an embodiment of the invention, the response value is obtained by detecting a change in cAMP concentration in the cell.
Illustratively, the change in cAMP concentration in the cells is obtained by using the glosensort (tm) cAMP detection kit. The GloSensor-20F cAMP gene construct can pre-express a luciferase variant, the increase of cAMP concentration can cause the conformational change of the luciferase variant, so that the luciferase is converted from an inactive state to an active state, and the GloSensor (TM) cAMP detection kit provides a substrate of the luciferase variant.
In a fifth aspect of the invention, a method of assessing the quality of a fragrance product is presented. According to an embodiment of the invention, the method comprises: contacting a perfume product to be detected with an olfactory receptor, and determining a response value of the olfactory receptor after the contact; determining the quality of the perfume product to be tested based on the response value; wherein the olfactory receptor comprises OR2J2 and/OR OR2W1. It can be known from the foregoing that 4-methoxybenzaldehyde can activate the olfactory receptor, so that the perfume product to be tested is contacted with the olfactory receptor, and if the perfume product to be tested contains 4-methoxybenzaldehyde, the olfactory receptor can be activated, and a response value after the olfactory receptor is activated is obtained, and the quality of the perfume product to be tested can be determined according to the magnitude of the response value. The perfume product to be tested can be a final perfume product or an intermediate product in the production of the perfume product, and when the final perfume product is the final perfume product, the final perfume product can be subjected to quality control according to the quality of the perfume product to be tested, and when the intermediate product is the final perfume product, the final perfume product can be used for knowing the production and preparation of the perfume product.
Herein, the term "fragrance product" refers to any product comprising plant fragrances (star anise, fennel, vanilla beans, corn and acacia), without limitation, such as spice powder, spice oil (e.g., spice essential oil or edible spice oil), and the like.
Illustratively, when the flavor product is a flavor essential oil, it includes, but is not limited to, essential oil products comprising at least one of fennel oil, star anise oil, acacia oil, and corn oil.
According to an embodiment of the invention, said contact is preceded by a dilution treatment of said perfume product to be tested.
Illustratively, the perfume product is a perfume powder, and the diluted mass-to-volume ratio of the perfume product is not less than 0.01g/ml, such as not less than 0.02g/ml, 0.03 g/ml, 0.04g/ml, 0.05 g/ml, 0.06g/ml, 0.07 g/ml, 0.08g/ml, and the like.
Illustratively, the flavor product is a flavor essential oil, and the flavor product has a diluted volume ratio of no less than 0.0004, e.g., no less than 0.0004, 0.0005, 0.0008, 0.0010, 0.0012, 0.0016, 0.0020, 0.0025, 0.0030, 0.0032, and the like.
According to an embodiment of the present invention, the perfume product to be tested contains at least one of star anise, fennel, vanilla beans, dill, corn and acacia.
In the present invention, "fennel" and "fennel" are synonymous.
In a preferred embodiment of the invention, the olfactory receptor is OR2J2.
According to an embodiment of the invention, said response value, being higher than a predetermined threshold value, is an indication that the perfume product to be tested is of acceptable quality.
It should be noted that the "predetermined threshold" can be obtained by detecting a large number of perfume products (for example, 20, 50, 100, 150, 200 or more) using the olfactory receptor of the present invention and counting the response values obtained from the large number of perfume products. Here, the "predetermined threshold" may be a response value directly obtained, may be a corresponding multiple with respect to a control group, and the specific type is not limited.
In some alternative embodiments of the invention, the method may be used to assess the quality of a fragrance product containing predetermined raw materials, for example, anise, fennel, vanilla beans, dill, corn and acacia.
Illustratively, the predetermined threshold value for evaluating the quality of the fennel-containing flavor product may be 50 to 60 μ g/1.2ml, or the molarity may be 300 to 400 μ M.
According to the embodiment of the invention, a plurality of perfume products to be tested are respectively contacted with olfactory receptors, and the response value of the olfactory receptors after the contact is determined; determining the quality of the perfume product to be tested based on the response value, wherein a higher response value is an indication of a higher quality of the perfume product to be tested.
For example, as shown in fig. 4, when there are two types of perfume products to be tested, the quality of the perfume product to be tested having a high response value (e.g., star anise oil produced by peasant in Guangxi) is better than the quality of the perfume product to be tested having a low response value (e.g., star anise oil produced by Yisheng.
According to an embodiment of the invention, the olfactory receptor is provided by a cell or transgenic cell expressing the olfactory receptor.
According to an embodiment of the invention, the cell or transgenic cell is a eukaryotic cell or a prokaryotic cell.
In some alternative embodiments of the invention, the eukaryotic cells include, but are not limited to, cells selected from the group of cells isolated from the olfactory baseplate, HEK293 cells, CHO cells, xenopus oocytes, hela cells, COS cells, yeast cells, and the like.
According to an embodiment of the invention, the prokaryotic cell is selected from bacteria.
According to an embodiment of the invention, said response value is obtained by detecting a change in activity of said olfactory receptor.
According to an embodiment of the invention, said activity change is determined by at least one of the following detection methods: luciferase assay, secreted alkaline phosphatase assay, fluorescent protein assay, fluorescent probe assay, ca 2+ Concentration detection, current detection, isotopic labeling, antibody detection, and pH detection.
Illustratively, the luciferase assay is a method in which when cells containing olfactory receptors are stimulated with 4-methoxybenzaldehyde or a sample containing 4-methoxybenzaldehyde, if the olfactory receptors are activated, the intracellular concentration of cAMP increases, cAMP binds to the promoter region of CRE-luciferase and promotes transcription and translation of luciferase, and thus, by detecting the activity of luciferase, the response of the olfactory receptors can be characterized, and it can be determined whether the olfactory receptors recognize 4-methoxybenzaldehyde.
According to an embodiment of the invention, said response value is obtained by detecting a change in cAMP concentration in said cell.
Illustratively, the change in cAMP concentration in the cells is obtained by using the glosensort (tm) cAMP detection kit. The GloSensor-20F cAMP gene construct can pre-express a luciferase variant, the increase of cAMP concentration can cause the conformational change of the luciferase variant, so that the luciferase is converted from an inactive state to an active state, and the GloSensor (TM) cAMP detection kit provides a substrate of the luciferase variant.
The scheme of the invention will be explained with reference to the following examples. It will be appreciated by those skilled in the art that the following examples are illustrative of the invention only and should not be taken as limiting the scope of the invention. The examples do not specify particular techniques or conditions, and are performed according to techniques or conditions described in literature in the art or according to the product specification. The reagents or instruments used are conventional products which are commercially available, and are not indicated by manufacturers.
Example 1: screening of olfactory receptors
In this example, the olfactory receptor activity is measured by the Dual-Luciferase method (Dual-Glo-Luciferase Assay System, promega). The inventors selected 398 olfactory receptors from the human olfactory receptor library, then prepared a genetic construct containing the olfactory receptors, golf, CRE-Luciferase and pRL-SV40, and transfected the genetic construct into HEK293T cells using the transfection reagent Lipofectamine2000 (Invitrogen). After 24 hours of culture, 4-methoxybenzaldehyde was diluted with a medium to a concentration of 300. Mu.M, and the cells were stimulated with diluted 4-methoxybenzaldehyde and incubated for 2 to 4 hours. If after the olfactory receptor is activated, the cAMP concentration in the cell is increased, the cAMP can bind to a CRE-luciferase promoter region and promote the transcription and translation of luciferase, and the response situation of the olfactory receptor can be characterized by detecting the activity of the luciferase, and finally 2 response-producing human olfactory receptors OR2J2 (Genbank ID:26707, protein ID.
The inventors also measured the stimulation of cells containing the olfactory receptors OR2J2 and OR2W1 by 4-methoxybenzaldehyde at different concentrations and observed the dose curves of the 2 olfactory receptors in response to 4-methoxybenzaldehyde at different concentrations, as shown in FIG. 2, in which the horizontal axis represents the logarithm of the concentration (M) of 4-methoxybenzaldehyde and the vertical axis represents the response change multiple relative to the blank control (without 4-methoxybenzaldehyde). As a result, it was found that OR2J2 responded most strongly to 4-methoxybenzaldehyde among the 2 olfactory receptors, and that the response of OR2J2 and OR2W1 varied by about 20-fold and 12-fold, respectively, and the sensitivity was 30. Mu.M and 100. Mu.M, respectively.
Example 2: detection of anise and fennel products by OR2J2 olfactory receptors
In this example, the method of example 1 was used to test the food seasonings for anise and fennel by cells expressing OR2J2. Star anise and fennel are the brand of huiying, and star anise and fennel are the solid, and the inventor extracts aromatic substance with water, specifically is: adding 20g of anise and fennel into 250ml of distilled water respectively, boiling for 20min, collecting the residual liquid to be fully filled to 24ml, and then diluting, wherein the specific dilution refers to the abscissa of figure 3; the diluted anise and fennel were then tested as in example 1. The results are shown in FIG. 3, wherein the horizontal axis represents the mass to volume ratio (in g/ml) and the vertical axis represents the response change fold relative to the blank (without anise and fennel).
The result shows that the content of the 4-methoxybenzaldehyde in the fennel of the Yingying brand is obviously higher than that of the anise of the Yingying brand, and when the content of the anise and the fennel is 0.08g/ml, the response multiple of the anise can reach about 7 times, and the response multiple of the fennel can reach about 32 times. Therefore, 4-methoxybenzaldehyde in the seasoning can be sensitively detected by using an OR2J2 olfactory receptor.
Example 3: detection of different brands of anise essential oil by OR2J2 olfactory receptor
In this example, 2 star anise essential oils were detected by cells expressing OR2J2 using the method of example 1. The 2 kinds of essential oil are produced by Guangxi farmhouse and plant Yisheng brand essential oil. The inventor uses ethanol to dilute the 2 kinds of essential oil to 1/4 respectively, and then dilutes the essential oil again by using a CD293 culture medium, and the specific dilution refers to the abscissa of FIG. 4; the diluted 2 anise essential oils were then tested as in example 1. The results are shown in FIG. 4, wherein the horizontal axis represents the volume ratio and the vertical axis represents the response change fold relative to the blank control (without adding anise oil).
As can be seen from fig. 4, when the ratio of the star anise oil is 0.0032, that is, when the star anise oil is diluted 320 times, the response multiple of the Guangxi farmhouse produced star anise oil reaches about 8 times, the response multiple of the plant Yisheng star anise oil reaches about 4 times, and the content of 4-methoxybenzaldehyde in the Guangxi farmhouse produced star anise oil is higher than that of the plant Yisheng star anise oil, which indicates that the Guangxi farmhouse produced star anise oil has better quality, and may be more conducive to retaining key aroma components in food due to the extraction process of the Guangxi farmhouse produced star anise oil. Thus, the use of the OR2J2 olfactory receptor can be used to assess the quality of commercially available essential oils.
Example 4: detection of fennel released gas by OR2J2 olfactory receptor
In this example, the method of example 1 was used to detect the gas released by fennel by cells expressing OR2J2. The method comprises the following specific steps: weighing 20g of fennel, placing the fennel in a sealed aluminum box, and after the fennel naturally releases fragrance for 12 hours, putting cells expressing OR2J2 olfactory receptors into the box for stimulation for 4 hours; and cells that did not contact the fragrance of fennel were set as blank controls. The detection result is shown in fig. 5, and the result shows that the response multiple of the cells which are placed in the aluminum box and express the OR2J2 olfactory receptor to the gas is about 3 times of that of the blank control group, and the result shows that the bionic olfactory recognition of the gas is possible by directly adopting the biological olfactory receptor cells.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples and features of different embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction.
Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.
Claims (10)
1. Use of olfactory receptors comprising OR2J2 and/OR 2W1 in the recognition of 4-methoxybenzaldehyde.
Use of 4-methoxybenzaldehyde to activate olfactory receptors comprising OR2J2 and/OR 2W1.
3. Use according to claim 1 or2, characterized in that said recognition or activation is represented by a change in the activity of olfactory receptors;
the activity change comprises at least one of the following signal changes:
luciferase, secreted alkaline phosphatase, fluorescent protein, fluorescent probe, cAMP, IP3, calcium ion, current, and pH.
4. A method for detecting 4-methoxybenzaldehyde, comprising:
contacting a sample to be detected with an olfactory receptor, and determining a response value of the olfactory receptor after the contact;
determining whether the sample to be detected contains 4-methoxybenzaldehyde or not based on the response value;
wherein the olfactory receptor comprises OR2J2 and/OR OR2W1.
5. The method of claim 4, wherein said olfactory receptor present response value is an indication that 4-methoxybenzaldehyde is contained in said test sample; or, the absence of a response value by the olfactory receptor is an indication that 4-methoxybenzaldehyde is not present in the sample to be tested;
the method further comprises the following steps:
and determining the content of the 4-methoxybenzaldehyde in the sample to be detected based on a standard curve, wherein the standard curve is a corresponding curve of a predetermined amount of the 4-methoxybenzaldehyde and the response value of the olfactory receptor.
6. A method of testing a food material, comprising:
contacting a sample to be detected with an olfactory receptor, and determining a response value of the olfactory receptor after the contact;
determining the raw material of the sample to be detected based on the response value;
wherein the olfactory receptor comprises OR2J2 and/OR OR2W1.
7. The method according to claim 6, wherein the olfactory receptor-present response value is an indication that the sample to be tested contains at least one of anise, fennel, vanilla beans, dill, corn, and acacia; alternatively, the first and second electrodes may be,
the absence of a response value by the olfactory receptor is an indication that the sample to be tested does not contain anise, fennel, vanilla beans, dill, corn, and acacia.
8. A method of assessing the quality of a fragrance product, comprising:
contacting a perfume product to be detected with an olfactory receptor, and determining a response value of the olfactory receptor after the contact;
determining the quality of the perfume product to be tested based on the response value;
wherein the olfactory receptor comprises OR2J2 and/OR OR2W1.
9. The method of claim 8, wherein the flavor product to be tested comprises at least one of aniseed, fennel, vanilla, corn, and acacia;
the response value is higher than a preset threshold value and is an indication that the quality of the perfume product to be detected is qualified;
respectively contacting a plurality of perfume products to be tested with olfactory receptors, and determining the response values of the olfactory receptors after the contact;
determining a quality of the perfume product to be tested based on the response value,
wherein a higher response value is an indication of a higher quality of the fragrance product to be tested.
10. The method of any one of claims 4~9 wherein said olfactory receptor is provided by a cell or transgenic cell that expresses said olfactory receptor;
said response value is obtained by detecting a change in cAMP concentration in said cell;
the cell or the transgenic cell is a eukaryotic cell or a prokaryotic cell;
said response value is obtained by detecting a change in activity of said olfactory receptor;
the change in activity is determined by at least one of the following detection methods:
luciferase assay, secreted alkaline phosphatase assay, fluorescent protein assay, fluorescent probe assay, ca 2+ Concentration detection, amperometric detection, isotopic labeling, antibody detection, and pH detection.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130281324A1 (en) * | 2010-04-16 | 2013-10-24 | Nuevolution A/S | Bi-functinal complexes and methods for making and using such complexes |
| CN104726551A (en) * | 2013-12-15 | 2015-06-24 | 西姆莱斯股份公司 | Method For Evaluating The Scent Performance Of Perfumes And Perfume Mixtures |
| CN109071626A (en) * | 2016-02-24 | 2018-12-21 | 阿罗姆斯公司 | For detecting smell, fragrance and the biosensor of taste |
| CN110214273A (en) * | 2016-09-30 | 2019-09-06 | 马斯公司 | Use the screening technique and the noval chemical compound identified in this way of olfactory receptor |
| US20200399558A1 (en) * | 2019-06-21 | 2020-12-24 | The Regents Of The University Of California | Methods for identifying, compounds identified and compositions thereof |
| US20210145768A1 (en) * | 2018-04-16 | 2021-05-20 | La Jolla Institute For Allergy And Immunology | Modulating immune response via targeting of olfactory receptor activity |
| CN114720645A (en) * | 2022-05-18 | 2022-07-08 | 汉王科技股份有限公司 | Use of olfactory receptor for recognizing 3-methylthiopropanal and method for detecting 3-methylthiopropanal |
-
2023
- 2023-02-13 CN CN202310140517.XA patent/CN115840026B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130281324A1 (en) * | 2010-04-16 | 2013-10-24 | Nuevolution A/S | Bi-functinal complexes and methods for making and using such complexes |
| CN104726551A (en) * | 2013-12-15 | 2015-06-24 | 西姆莱斯股份公司 | Method For Evaluating The Scent Performance Of Perfumes And Perfume Mixtures |
| US20150260707A1 (en) * | 2013-12-15 | 2015-09-17 | Symrise Ag | Method for evaluating the scent performance of perfumes or perfume mixtures |
| CN109071626A (en) * | 2016-02-24 | 2018-12-21 | 阿罗姆斯公司 | For detecting smell, fragrance and the biosensor of taste |
| CN110214273A (en) * | 2016-09-30 | 2019-09-06 | 马斯公司 | Use the screening technique and the noval chemical compound identified in this way of olfactory receptor |
| US20210145768A1 (en) * | 2018-04-16 | 2021-05-20 | La Jolla Institute For Allergy And Immunology | Modulating immune response via targeting of olfactory receptor activity |
| US20200399558A1 (en) * | 2019-06-21 | 2020-12-24 | The Regents Of The University Of California | Methods for identifying, compounds identified and compositions thereof |
| CN114720645A (en) * | 2022-05-18 | 2022-07-08 | 汉王科技股份有限公司 | Use of olfactory receptor for recognizing 3-methylthiopropanal and method for detecting 3-methylthiopropanal |
Non-Patent Citations (1)
| Title |
|---|
| TAO TONG 等: "Ectopic Odorant Receptor Responding to Flavor Compounds: Versatile Role in Health and Disease", PHARMACEUTICS * |
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