KR101719718B1 - Apparatus and method for measuring ripening degree using light-emitting microorganisms - Google Patents

Abstract

According to an aspect of the present invention, there is provided a method of measuring a biological sample, comprising the steps of: (a) (b) reacting the gas sample with a luminescent microorganism culture solution; And (c) comparing the light emission intensity before and after the reaction of the light emitting microorganism.
According to another aspect of the present invention, there is provided a gas sampling apparatus including: a sample inlet for sucking a gas sample outside an object to be examined; A reaction part including a culture medium for luminescent microorganisms connected to the sample inlet part; A sensing unit for measuring the intensity of the light emitted from the reaction unit and converting the measured intensity into an electric signal; A display unit for outputting a light change amount based on the electrical signal; And an operating unit for applying a suction force to the sample inlet unit.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method and apparatus for measuring the degree of aging using light emitting microorganisms,

The present invention relates to an apparatus and a method for measuring the degree of maturity of fruit by comparing the luminescence activity before and after a reaction between a gas sample outside the subject and a luminescent bacteria, .

It is important to measure the maturity and quality of fruits and vegetables before harvesting because the quality of harvested produce and the final product sold to consumers must match. However, the degree of aging of fruits and vegetables is difficult to measure, especially when the color of fruits and vegetables is not related to aging. In addition, the aging of fruits and vegetables is judged by experts, but it can not be a realistic solution for testing large quantities of products.

Conventional methods for determining the degree of aging of fruits generally measure the hardness of fruits and vegetables using invasion or impact forces, or measure chemical levels and parameters (pH, titratable acidity, and saccharic acid content) associated with aging . However, these methods are destructive and can hinder the commerciality of fruits and vegetables.

In recent years, non-destructive methods for measuring the degree of fruit and vegetable aging include NMR (Nuclear Magnetic Resonance) and PMR (Proton Magnetic Resonance). These methods measure the saccharide acid level and use a vision system to measure fruit and vegetables And the hardness of fruits and vegetables is measured using an acoustic method. However, these methods require expensive equipment and may not achieve a high level of accuracy due to the inconsistent color and aging of fruits and vegetables.

Therefore, there is a demand for a technique that can confirm the aging of fruits and vegetables by a reliable, economical, and non-destructive method, easily readable by the general public without professional knowledge, and satisfactory to consumers, producers and distributors.

Disclosure of Invention Technical Problem [8] The present invention is intended to solve the problems of the prior art described above, and it is an object of the present invention to provide an apparatus and a method capable of measuring the degree of aging of fruits in an economical and non-destructive manner.

In order to achieve the above object, one aspect of the present invention provides a method of measuring a biological sample, comprising the steps of: (a) (b) reacting the gas sample with a luminescent microorganism culture solution; And (c) comparing the light emission intensity before and after the reaction of the light emitting microorganism.

In one embodiment, the gas sample may comprise ethylene gas.

In one embodiment, the light emitting microorganism is selected from the group consisting of Vibrio fischeri ), Photobacterium forum phosphoreum , or a recombinant microorganism transformed to enable luminescence.

In order to accomplish the above object, another aspect of the present invention provides a sample injection apparatus including: a sample inlet for sucking a gas sample outside an object to be examined; A reaction part including a culture medium for luminescent microorganisms connected to the sample inlet part; A sensing unit for measuring the intensity of the light emitted from the reaction unit and converting the measured intensity into an electric signal; A display unit for outputting a light change amount based on the electrical signal; And an operating unit for applying a suction force to the sample inlet unit.

In one embodiment, the gas sample may comprise ethylene gas.

In one embodiment, the light emitting microorganism is selected from the group consisting of Vibrio fischeri ), Photobacterium forum phosphoreum , or a recombinant microorganism transformed to enable luminescence.

In one embodiment, the reaction part may include a sample storage part and a microorganism storage part into which the light emitting microorganism culture solution is injected.

In one embodiment, the outer wall of the sample storage part may be a semipermeable membrane.

In one embodiment, the sensing unit may be a photodiode, a phototransistor, a photothyristor, a CCD (Charge Coupled Device) image sensor, or a CMOS (Complementary Metal-Oxide Semiconductor) have.

The present invention provides an economical and effective method for measuring the degree of maturation without physical damage to the subject.

It should be understood that the effects of the present invention are not limited to the above effects and include all effects that can be deduced from the detailed description of the present invention or the configuration of the invention described in the claims.

FIG. 1 is a schematic diagram of an apparatus for measuring the degree of fruit maturity according to an embodiment of the present invention.
2 is a schematic view illustrating a structure of a reaction part according to an embodiment of the present invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described with reference to the accompanying drawings. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. In order to clearly illustrate the present invention, parts not related to the description are omitted, and similar parts are denoted by like reference characters throughout the specification.

Throughout the specification, when a part is referred to as being "connected" to another part, it includes not only "directly connected" but also "indirectly connected" . Also, when an element is referred to as "comprising ", it means that it can include other elements, not excluding other elements unless specifically stated otherwise.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings.

In order to achieve the above object, one aspect of the present invention provides a method of measuring a biological sample, comprising the steps of: (a) (b) reacting the gas sample with a luminescent microorganism culture solution; And (c) comparing the light emission intensity before and after the reaction of the light emitting microorganism.

The fruit refers to the fruit of an edible plant, and the kind thereof is not particularly limited, and may include all kinds of ordinary fruits that are widely cultivated and circulated.

Typically, the fruit is able to synthesize ethylene when it is aged or when prospective cell death occurs. Ethylene is a kind of plant hormone, which is produced by using L-methionine as a precursor and using S-adenosyl methionine, 1-aminocyclo propane-1-carboxylic acid acid, and finally synthesized by 1-aminocyclopropane-1-carboxylic acid oxidase.

The ethylene (CH ₂ CH ₂ ) is a kind of plant hormone and may exist in a gaseous state in all plant tissues, and ethylene synthesized by aging may diffuse out of the plant tissue. The ethylene promotes the decomposition of chlorophyll and induces the synthesis of antioxidants or carotenoids to change the color of the fruit to red or a predetermined color and promote the fruiting and softening of the fruit .

The amount of ethylene produced varies depending on the type of fruit, but increases before and after harvest, and mature fruit can produce a large amount of ethylene. Particularly, when the concentration of ethylene reaches 0.1 to 1.0 ppm, the fruit may be aged excessively and the tissue may become torn.

Since the ethylene is present in a gaseous state in all plant tissues, it acts as a growth hormone or a mature hormone to promote fruit respiration and aging, thereby improving the coloring of fruit vegetables. Therefore, the amount of ethylene released from the fruits is quantified, The degree of aging can be evaluated.

Accordingly, the method for measuring the degree of fruit maturity comprises the steps of: (a) sampling a gas sample outside the subject to be examined; (b) reacting the gas sample with a luminescent microorganism culture solution; And (c) comparing light emission intensity before and after the reaction of the light emitting microorganism.

The subject to be tested may be ordinary fruit to be subjected to the measurement of the degree of maturity, but is not limited thereto, and may include all plants such as deep-fried vegetables, root vegetables, fruit vegetables, green vegetables, and flowers.

Since ethylene produced in the tissue by aging can be diffused out of the air, the gas sample and the light emitting microorganism are collected from the test subject outside the test subject containing the ethylene, and the ethylene is easily quantified can do.

That is, the fruit to be tested releases ethylene to the outside due to aging. When the external gas sample and the light emitting microorganism culture liquid come into contact with each other, the ethylene contained in the gas sample can be dissolved in the culture solution.

At this time, the ethylene may be dissolved in the culture solution and converted to ethylene oxide. The ethylene oxide is very biochemically highly reactive, which causes damage to the nervous system of the organism and can adversely affect the biological activity of aquatic organisms.

Therefore, it is possible to easily quantify the ethylene release amount of the test subject by collecting the gas sample outside the subject and reacting with the light emitting microorganism, and the degree of maturation of the test subject can be measured based on the result. The luminescent microorganisms are vulnerable to toxic substances and react very sensitively. Therefore, it is possible to rapidly detect toxic substances, and the cost for cultivation is low.

On the other hand, the light-emitting microorganism is a microorganism that emits light by a biological mechanism, and the kind thereof is not limited, and preferably Vibrio fischeri ), Photobacterium forum phosphoreum , or a recombinant microorganism transformed to enable luminescence.

Specifically, the light emitting microorganism is a microorganism inhabited mainly in the ocean, and can emit visible light of a wavelength band of about 400 nm in a metabolic process. The light-emitting microorganism may be weakened or stopped because it is killed or the vitality is lowered when exposed to a toxic substance. It is reported that the bioluminescence emits light of a specific region by luciferase in a cell oxidizes a reduced flavin mononucleotide (FMNH ₂ ) and a long chain aliphatic aldehyde.

The recombinant microorganism is Gene coding for the gene (lux gene) or a fluorescent protein coding for the light-emitting enzyme may retain luminescent activity. If you can emit visible light to identify whether the light emission that the type is not limited Do not.

The fluorescent protein may be selected from the group consisting of a red fluorescent protein (RFP), an enhanced red fluorescent protein (ERFP), a green fluorescent protein (GFP), a modified green fluorescent protein, (EFP), a blue fluorescent protein (BFP), an enhanced blue fluorescent protein (EBFP), a cyan fluorescent protein (CFP), an enhanced cyan fluorescent protein (ECFP), a yellow fluorescent protein Type protein (EYFP), but the present invention is not limited thereto.

On the other hand, the microorganism culture solution can be standardized by injecting the colony of the optimal density into the culture solution in which the light-emitting microorganism has sufficiently dissolved nutrients. Since the luminescent phenomenon can be easily observed when the luminescent microorganism is a colony, the density of microorganisms can be appropriately controlled so that the sensitivity and accuracy can be maximized.

In one embodiment, the microbial culture may be an optimized medium selected from the Plackett-Burman experimental design method or the 2 factor central composite method, It is possible to control the density of microorganisms in the culture liquid.

For example, the Eggplug-Burman experimental design method is a method for selecting a main culture medium of luminescent microorganisms, which comprises a main substrate such as tryptone, yeast extract, glycerol, sodium chloride, potassium chloride, CaCl ₂ , H ₂ O, MgCl ₂ The effects of H ₂ O, NaHCO ₃ , and MgSO ₄ .H ₂ O on the bioluminescence and growth of the luminescent microorganisms are analyzed, and the optimal concentration can be calculated by the two factor central method.

Since the amount of ethylene produced by the aging of the subject can be different depending on the kind of the subject to be examined, the difference in the amount of light emission according to the amount of light emission according to the optimal concentration of the microorganism and the degree of aging of the subject can be set in advance. It is possible to determine the degree of aging for each test subject based on the preset data. For example, since the amounts of ethylene produced by aging of apples and bananas are different, the amount of change in the luminescence intensity of the light emitting microorganisms according to the degree of aging of apples can be measured and data can be measured in advance, and data on bananas can also be separately data. That is, the degree of maturation of the test subject can be easily evaluated by previously data of the degree of light emission of the light emitting microorganisms according to the type of the test subject.

The light emission intensity of the light emitting microorganisms before and after the reaction can be measured using a conventional photosensor capable of electronically sensing the light amount, but it is not particularly limited as long as the light amount can be accurately measured have.

According to another aspect of the present invention, there is provided a sample injection apparatus comprising: a sample inlet portion for sucking a gas sample outside an object to be tested; A reaction part 200 connected to the sample inlet part 100 and including a culture medium for luminescent microorganisms; A sensing unit 300 for measuring the intensity of light emitted from the reaction unit 200 and converting the measured intensity into an electrical signal; A display unit 400 for outputting a light change amount based on the electrical signal; And an operating part (500) for applying a suction force to the sample inlet part (100).

Referring to FIG. 1, the fruit maturity measuring apparatus may include a sample inlet 100, a reaction unit 200, a sensing unit 300, a display unit 400, and an operation unit 500. The above-mentioned fruit aging measuring apparatus can measure the degree of aging of fruits by taking a gas sample outside the fruit to be tested.

In one embodiment, the sample inlet 100 may be located at one side of the fruit maturity measuring device and may have a conical shape in which the lower end is gradually narrowed, The bonding method or form is not particularly limited as long as it is a structure in which the gas sample can be effectively introduced at the location of the gas sample.

The manipulation part 500 may apply a suction force to the sample inlet part 100.

According to one embodiment, the manipulation part 500 may have a button-like structure located on the other side of the sample inlet part 100, and may be coupled with a spring device to hold a predetermined elastic force. The operating portion 500 can be lowered by pressure. When the operating portion 500 is lowered, the internal pressure is increased, so that the internal gas sample can be discharged to the outside. In addition, the operating unit 500 can be raised by the spring device when the pressure is re-applied. Since the pressure inside the operating unit 500 is reduced when the operating unit 500 is raised, a suction force is applied to the sample inlet 100 , An external gas sample can be sucked.

However, the operating part 500 is not particularly limited as long as it is capable of providing a suction force to the sample inlet part 100. [ For example, the operation unit 500 may be a soft rubber or silicon material capable of being compressed and restored, and may be sucked in an external gas sample by restoration after compression, or may be an electronic device having a motor and a battery.

The reaction part 200 may be adjacent to the sample inlet part 100 and may react with the gas sample drawn into the sample inlet part 100.

2, the reaction unit 200 may include a sample storage unit 210 in which the inhaled gas sample is submerged for a predetermined time, and a microorganism storage unit 220 into which the microorganism culture is injected. The outer wall of the sample storage part 210 may be a semi-permeable membrane.

In one embodiment, the sample storage unit 210 and the microorganism storage unit 220 may be in contact with each other.

The semi-permeable membrane 211 may include fine pores of 5 μm or less, and may be an organic material or an inorganic material. The semi-permeable membrane 211 may be formed of a material selected from the group consisting of cellulose acetate, cellulose nitrate, polysulfone, polyvinylidene fluoride, polyamide, and acrylonitrile. Any material that can be a copolymer but possesses semi-permeable properties can be used without limitation.

Since the semi-permeable membrane 211 can permeate the solvent molecules or ions of a specific size or less, water, water vapor, and other solid foreign substances can not be discharged to the outside, but the sample storage unit (not shown) adjacent to the microorganism storage unit 220 210 may be introduced into the microorganism storage part 220 through the semi-permeable membrane 211.

Therefore, the ethylene contained in the gas sample is dissolved in the culture solution of the light emitting microorganism and converted into ethylene oxide, and the metabolism of the light emitting microorganism is inhibited by the toxicity of the ethylene oxide, so that the light emission intensity of the light emitting microorganism can be reduced.

Referring to FIG. 2, the sample storage unit 210 may have a narrow cylindrical shape, and the microorganism storage unit 220 may surround the outer surface of the sample storage unit 210.

Therefore, the gas sample taken in the sample inlet part 100 may be located in the sample storage part 210, and the gas sample may be passed through the semi-permeable membrane 211 in contact with the sample storage part 210 And may be introduced into the microorganism storage part 220. The shape of the sample storage part 210 and the microorganism storage part 220 is not particularly limited and is not particularly limited as long as the gas sample can move from the adjacent contact surface.

In addition, the microorganism storage part 220 may include a microorganism culture solution containing sufficient nutrients. However, the nutrients contained in the microorganism storage part 220 are continuously consumed due to the growth of the microorganisms and can not be used permanently And may be a removable structure that can be replaced after use. Therefore, the microorganism storage part 220 may contain an optimum density of the light emitting microorganism culture solution, and may be replaced with a new microorganism storage part 220 after use and reused.

The sensing unit 300 may include an optical sensor for measuring the intensity of the light. The sensing unit 300 may measure the intensity of light emitted from the microorganism storage unit 220, convert the intensity of the light into electrical signals, and transmit the electrical signal to the display unit 400 .

The sensing unit 300 may be a photodiode, a phototransistor, a photothyristor, a CCD (Charge Coupled Device) image sensor, a CMOS (Complementary Metal-Oxide Semiconductor) Image sensors, and different types of sensors may be combined to improve sensitivity and accuracy.

Photodiodes, phototransistors, and photothyristors generate free electrons and free-hole pairs when a photon having sufficient energy enters a pn junction or a pin junction to excite electrons. The carrier becomes a photocurrent, and the diode characteristic can be changed by the generated photocurrent, or the transistor or the thyristor can be operated.

The CCD (Charge Coupled Device) image sensor generates a charge when light is incident on the light receiving element, and the charge can be detected by transmitting the charge using a circuit element called a charge coupled device (CCD: Charge Coupled Device) .

The complementary metal-oxide semiconductor (CMOS) image sensor converts the charge accumulated in the photodiode, which is a light-receiving element, into a voltage in each pixel to convert the light into an electrical signal. Since the CMOS image sensor is small and consumes less power, it is advantageous for miniaturization.

The sensing unit 300 can directly measure a change in the amount of light of the reaction unit 200 or receive the light through the optical fiber. The sensing unit 300 may convert a change in light amount into a charge amount and analyze and store the charge amount. In addition, the sensing unit 300 may further include an operation unit for calculating the concentration of ethylene based on the charge amount data.

The change in the amount of charge can be analyzed by a magic magic, a specific velocity method, and a bioluminescence method, and the concentration of ethylene contained in the gas sample can be calculated by the analysis.

In one embodiment, the display unit 400 may output a light change amount emitted by the light emitting microorganisms based on an electrical signal of the sensing unit 300.

The display unit 400 may be any type that allows the user to easily recognize the degree of maturation of the subject, for example, a liquid crystal display (LCD), a flat panel display (PDP) Emitting Diode) or AMOLED (Active Matrix Organic Light Emitting Diode) display device, but the type is not limited and it is sufficient if information can be transmitted to the user through various means such as sound or light.

It will be understood by those skilled in the art that the foregoing description of the present invention is for illustrative purposes only and that those of ordinary skill in the art can readily understand that various changes and modifications may be made without departing from the spirit or essential characteristics of the present invention. will be. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive. For example, each component described as a single entity may be distributed and implemented, and components described as being distributed may also be implemented in a combined form.

The scope of the present invention is defined by the appended claims, and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be construed as being included within the scope of the present invention.

100: Sample inlet
200: reaction part
210:
211: Semi-permeable membrane
220: Microorganism reservoir
300:
400:
500:

Claims (9)

A sample inlet for sucking a gas sample outside the object to be examined;
A reaction part connected to the sample inlet part and including a sample storage part and a microorganism storage part into which a culture solution of the light emitting microorganism is injected;
A sensing unit for measuring the intensity of the light emitted from the reaction unit and converting the measured intensity into an electric signal;
A display unit for outputting a light change amount based on the electrical signal; And
And an operating part for applying a suction force to the sample inlet part,
Wherein the light emitting microbe reacts with ethylene gas produced by the fruit to reduce the luminescence intensity.
delete The method according to claim 1,
The light emitting microorganism Vibrio blood Sherry (Vibrio fischeri ), Photobacterium forum phosphoreum , or a recombinant microorganism transformed to enable luminescence.
delete The method according to claim 1,
Wherein the outer wall of the sample storage part is made of a semi-permeable membrane.
The method according to claim 1,
Characterized in that the sensing part comprises a photodiode, a phototransistor, a photothyristor, a CCD (Charge Coupled Device) image sensor, or a CMOS (complementary metal-oxide semiconductor) Aging degree measuring device.
(a) collecting a gas sample outside the fruit;
(b) reacting the gas sample with a luminescent microbial culture capable of reacting with ethylene; And
(c) comparing the light emission intensity before and after the reaction of the light emitting microorganism.
delete 8. The method of claim 7,
When the light-emitting microorganism is a Vibrio fissure (Vibrio fischeri), Photo Bacterium Force ForumPhotobacterium 포스코 호), Or a recombinant microorganism transformed to enable light emission.

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