[go: up one dir, main page]

WO2018147443A1 - Kit permettant de mesurer le degré de contamination d'instruments de cuisson et procédé d'évaluation d'un programme de nettoyage d'instruments de cuisson - Google Patents

Kit permettant de mesurer le degré de contamination d'instruments de cuisson et procédé d'évaluation d'un programme de nettoyage d'instruments de cuisson Download PDF

Info

Publication number
WO2018147443A1
WO2018147443A1 PCT/JP2018/004722 JP2018004722W WO2018147443A1 WO 2018147443 A1 WO2018147443 A1 WO 2018147443A1 JP 2018004722 W JP2018004722 W JP 2018004722W WO 2018147443 A1 WO2018147443 A1 WO 2018147443A1
Authority
WO
WIPO (PCT)
Prior art keywords
atp
program
adp
cooking
kit
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/JP2018/004722
Other languages
English (en)
Japanese (ja)
Inventor
有紀 塚田
繁哉 鈴木
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Kikkoman Corp
Original Assignee
Kikkoman Corp
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Kikkoman Corp filed Critical Kikkoman Corp
Priority to JP2018567526A priority Critical patent/JP7227008B2/ja
Publication of WO2018147443A1 publication Critical patent/WO2018147443A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
    • C12Q1/42—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving phosphatase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/48—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/66—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving luciferase
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/76—Chemiluminescence; Bioluminescence

Definitions

  • the present invention relates to a kit for measuring the degree of contamination of cooking-related utensils and a method for measuring the degree of contamination using the kit. Moreover, this invention relates to the evaluation method of the cooking related utensil washing
  • SOP Standard Operating Procedure
  • SOPs and SSOPs for food processing, it is necessary to assess the degree of contamination of cooking-related utensils and confirm cleaning.
  • SSOP include (A) cleaning and sterilization of machinery and equipment and (B) cleaning and sterilization of workers' hands.
  • Patent Document 1 Japanese Patent Laid-Open No. 9-234099, Japanese Patent No. 34099962
  • Patent Document 2 Japanese Patent Laid-Open No. 11-069997
  • Non-Patent Document 1 Japanese Patent Laid-Open No. 11-069997
  • Japanese Patent Laid-Open No. 9-234099 Japanese Patent No. 3409996
  • Enzyme reagents that measure nucleotides such as ATP, ADP, and AMP are known, and these were available in laboratories and laboratories, but samples from individual cooking facilities were transported to laboratory facilities and research facilities one by one. It is expensive and time consuming to make measurements and communicate the results to the cooking site to modify the cleaning program. Moreover, the dishwasher is evaluated by the manufacturer at the time of development, but the evaluation method is not quantitative, such as qualitative or visual. In addition, there has never been a means for accurately and simply evaluating the degree of contamination when it is used at each business office or cooking site.
  • This invention makes it a subject to provide the cooking related appliance contamination degree measuring kit and cooking related appliance washing program evaluation method which solve the conventional problem.
  • the present invention provides an ATP and ADP measurement kit or an ATP, ADP and AMP measurement kit that can quickly detect contamination of cooking-related utensils, and a cleaning program evaluation using this kit. Provide a method.
  • the present invention includes the following embodiments: [1] Cleaning program evaluation method using ATP and ADP measurement kit, including the following steps: (i) Cleaning cooking-related utensils according to the program; (ii) Use the kit to measure the degree of contamination of washed cooking-related utensils. (iii) Compare the pollution degree measured in step (ii) with the reference specified in the program. (iv) If the measured pollution degree exceeds the reference specified in the program, Evaluate that the cleaning process of i) does not sufficiently remove contamination.
  • the ATP and ADP measurement kit is an ATP and ADP measurement kit for improving a cooking-related utensil washing program, which includes an enzyme that catalyzes a reaction for generating ATP from ADP, luciferin, luciferase, and a metal salt.
  • Cleaning program improvement method using ATP and ADP measurement kit including the following steps, (i) Cleaning cooking-related utensils according to the program, (ii) Use the kit to measure the degree of contamination of washed cooking-related utensils.
  • the ATP and ADP measurement kit is an ATP and ADP measurement kit for improving a cooking-related utensil washing program, which includes an enzyme that catalyzes a reaction for generating ATP from ADP, luciferin, luciferase, and a metal salt.
  • step (o) creating a cleaning program for cooking-related utensils;
  • Enzymes that catalyze the reaction of generating ATP from ADP are pyruvate kinase (PK), acetate kinase (AK), creatine kinase (CK), polyphosphate kinase (PPK), hexokinase, glucokinase, glycerol kinase, 5.
  • PK pyruvate kinase
  • AK acetate kinase
  • CK creatine kinase
  • PPK polyphosphate kinase
  • hexokinase glucokinase
  • glycerol kinase 5.
  • the method according to any one of 1 to 4 which is selected from the group consisting of fructokinase, phosphofructokinase, riboflavin kinase, and fructose bisphosphatase.
  • the ATP and ADP measurement kit further contains an AMP measurement reagent.
  • the AMP measurement reagent comprises an enzyme that catalyzes a reaction for producing ADP or ATP from AMP.
  • a cleaning program evaluation method using an ATP, ADP and AMP measurement kit including the following steps: (i) cleaning cooking-related utensils according to the program; (ii) Use the kit to measure the degree of contamination of washed cooking-related utensils. (iii) Compare the pollution degree measured in step (ii) with the reference specified in the program.
  • the measurement kit includes an enzyme that catalyzes a reaction that generates ATP from AMP, an enzyme that catalyzes a reaction that generates AMP from ADP, luciferin, luciferase, and a metal salt. Said method. [10] The method according to 9, wherein when it is evaluated that the contamination is not sufficiently removed in step (iv), the cooking-related utensils are further washed by the same or different method as in step (i).
  • a cleaning program improvement method using an ATP, ADP and AMP measurement kit including the following steps: (i) cleaning cooking-related utensils according to the program; (ii) Use the kit to measure the degree of contamination of washed cooking-related utensils. (iii) Compare the pollution degree measured in step (ii) with the reference specified in the program.
  • the measurement kit includes an enzyme that catalyzes a reaction that generates ATP from AMP, an enzyme that catalyzes a reaction that generates AMP from ADP, luciferin, luciferase, and a metal salt. Said method.
  • step (o) creating a cleaning program for cooking-related utensils comprising: [13]
  • the enzyme that catalyzes the reaction that generates ATP from AMP is pyruvate orthophosphate dikinase (PPDK) or pyruvate water dikinase (PWDK), and the enzyme that catalyzes the reaction that generates AMP from ADP.
  • PPDK pyruvate orthophosphate dikinase
  • PWDK pyruvate water dikinase
  • the method according to any one of 9 to 12 which is ADP-dependent hexokinase or apyrase.
  • ATP and ADP measurement kit for use in a cooking-related utensil washing program evaluation method, comprising an enzyme that catalyzes a reaction for generating ATP from ADP, luciferin, luciferase, and a metal salt, wherein the cooking-related utensil washing program is evaluated
  • the method is (i) cleaning cooking-related utensils according to the program; (ii) Use the kit to measure the degree of contamination of washed cooking-related utensils. (iii) compare the degree of contamination measured in step (ii) with the references specified in the program; and (iv) If the measured degree of contamination exceeds the reference specified in the program, evaluate that the cleaning process in (i) does not remove the contamination sufficiently.
  • a kit comprising the kit.
  • ATP and ADP measurement kit for use in a cooking-related utensil washing program improvement method, comprising an enzyme that catalyzes a reaction for generating ATP from ADP, luciferin, luciferase and a metal salt, wherein the cooking-related utensil washing program is improved
  • the method is (i) cleaning cooking-related utensils according to the program; (ii) Use the kit to measure the degree of contamination of washed cooking-related utensils.
  • step (iii) Compare the pollution degree measured in step (ii) with the reference specified in the program. (iv) If the measured pollution degree exceeds the reference specified in the program, modify the cleaning process of i), and (v) Repeat steps (i) to (iv) until the pollution level is below the reference specified in the program, A kit comprising the kit.
  • step (o) creating a cleaning program for cooking-related utensils comprising: [18] Enzymes that catalyze the reaction of generating ATP from ADP are pyruvate kinase (PK), acetate kinase (AK), creatine kinase (CK), polyphosphate kinase (PPK), hexokinase, glucokinase, glycerol kinase, The kit according to any one of 14 to 17, which is selected from the group consisting of fructokinase, phosphofructokinase, riboflavin kinase, and fructose bisphosphatase.
  • PK pyruvate kinase
  • AK acetate kinase
  • CK creatine kinase
  • PPK polyphosphate kinase
  • hexokinase hexokinase
  • glucokinase g
  • kits according to any one of 14 to 18, wherein the kit further contains an AMP measurement reagent.
  • the AMP measurement reagent comprises an enzyme that catalyzes a reaction for producing ADP or ATP from AMP.
  • the enzyme according to 20, wherein the enzyme that catalyzes a reaction for generating ADP or ATP from AMP comprises pyruvate orthophosphate dikinase (PPDK), adenylate kinase (ADK), or pyruvate water dikinase (PWDK).
  • PPDK pyruvate orthophosphate dikinase
  • ADK adenylate kinase
  • PWDK pyruvate water dikinase
  • ATP for use in cooking-related utensil washing program evaluation method, including enzyme that catalyzes reaction to produce ATP from AMP, enzyme that catalyzes reaction to produce AMP from ADP, luciferin, luciferase and metal salt, ADP And the AMP measurement kit, wherein the cooking-related utensil cleaning program evaluation method is: (i) cleaning cooking-related utensils according to the program; (ii) Use the kit to measure the degree of contamination of washed cooking-related utensils.
  • kits comprising the kit. [23] The kit according to 22, wherein when it is evaluated that the contamination is not sufficiently removed in step (iv), the cooking-related utensils are further washed by the same or different method as in step (i).
  • ATP ATP
  • ADP for use in cooking-related utensil cleaning program improvement method, including enzyme that catalyzes reaction for generating ATP from AMP, enzyme that catalyzes reaction for generating AMP from ADP, luciferin, luciferase, and metal salt
  • an AMP measurement kit wherein the cooking-related utensil cleaning program improvement method is: (i) cleaning cooking-related utensils according to the program; (ii) Use the kit to measure the degree of contamination of washed cooking-related utensils. (iii) Compare the pollution degree measured in step (ii) with the reference specified in the program.
  • kits comprising the kit.
  • the enzyme that catalyzes the reaction that generates ATP from AMP is pyruvate orthophosphate dikinase (PPDK) or pyruvate water dikinase (PWDK), and the enzyme that catalyzes the reaction that generates AMP from ADP.
  • PPDK pyruvate orthophosphate dikinase
  • PWDK pyruvate water dikinase
  • the kit according to any of 22 to 25 which is ADP-dependent hexokinase or apyrase.
  • the degree of contamination of cooking-related utensils can be measured without depending on visual observation.
  • special inspections can be performed easily and quickly especially for lots that require safety confirmation.
  • the problems of the conventional hygiene management program can be identified more accurately, and the program evaluation can be performed effectively.
  • ATP and ADP, or ATP is decomposed by heat, pH, time or ATP-degrading enzyme, or ATP and ADP, ATP, AMP, and ADP can be accurately measured, and the cleaning program can be effectively evaluated, a reference value can be set, and the cleaning program can be modified.
  • the reagents of the present invention are in a kit, it is not necessary to transport samples to a research facility one by one, and contamination can be detected quickly at each cooking site or cooking facility (in situ). And can quickly evaluate cleaning programs.
  • the present invention provides a cooking-related appliance contamination degree measuring kit and a cooking-related appliance contamination degree measuring method using the same.
  • the method uses an enzyme, luciferin, luciferase and a metal salt that catalyses the reaction of producing ATP from ADP, or a kit comprising these.
  • Enzymes that catalyze the reaction of generating ATP from the ADP include pyruvate kinase (PK), acetate kinase (AK), creatine kinase (CK), polyphosphate kinase (PPK), hexokinase, glucokinase, glycerol kinase, fructose It can be selected from the group consisting of kinase, phosphofructokinase, riboflavin kinase, and fructose bisphosphatase.
  • PK pyruvate kinase
  • AK acetate kinase
  • CK creatine kinase
  • PPK polyphosphate kinase
  • hexokinase hexokinase
  • glucokinase glucokinase
  • glycerol kinase glycerol kinase
  • the method further uses or uses a kit comprising pyruvate orthophosphate dikinase (PPDK), adenylate kinase or pyruvate water dikinase (PWDK).
  • PPDK pyruvate orthophosphate dikinase
  • PWDK pyruvate water dikinase
  • an enzyme that catalyzes a reaction that produces ATP from AMP an enzyme that catalyzes a reaction that produces AMP from ADP, luciferin, luciferase, and a metal salt are used, or a kit containing these is used.
  • the enzyme that catalyzes the reaction of generating ATP from the AMP can be selected from the group consisting of pyruvate orthophosphate dikinase (PPDK) and pyruvate water dikinase (PWDK), and generates AMP from the ADP.
  • PPDK pyruvate orthophosphate dikinase
  • PWDK pyruvate water dikinase
  • the enzyme that catalyzes the reaction may be selected from the group consisting of ADP-dependent hexokinase and apyrase.
  • the sample When the sample contains ATP, it is converted into AMP by luciferase and luminescence occurs.
  • ADP is contained in the sample, it is converted to ATP by an enzyme that catalyzes a reaction that generates ATP from ADP, and then ATP is subjected to a luminescence reaction.
  • the total amount of ATP and ADP present in the system can be measured.
  • PPDK if AMP is contained in the sample, this is converted to ATP by PPDK, PEP, and PPi.
  • PWDK if AMP is contained in the sample, this is converted to ATP by PWDK, PEP, and phosphoric acid.
  • the generated ATP again emits light by luciferase.
  • Luminescence is maintained stably, and the amount of luminescence correlates with the total amount of ATP and AMP present in the system, so that ATP and AMP can be quantified.
  • an enzyme that catalyzes the reaction of generating ATP from ADP and PPDK, ADK, or PWDK the total amount of ATP, ADP, and AMP can be measured.
  • the advantage of the method using PPDK or the like is that, even in a low-sensitivity apparatus, AMP produced by luciferase is also converted to ATP, so that luminescence can be measured stably without attenuation of luminescence.
  • the sample contains AMP and ATP, it is converted to 2 molecules of ADP by adenylate kinase.
  • the resulting ADP molecule can then be converted to ATP by an enzyme that catalyzes a reaction that produces ATP from ADP.
  • the resulting ATP can then be detected by luciferase.
  • an enzyme that catalyzes a reaction that generates ATP from AMP an enzyme that catalyzes a reaction that generates AMP from ADP, luciferin, luciferase, and a metal salt
  • ADP converts AMP from ADP. It can be converted to AMP by an enzyme that catalyzes the resulting reaction.
  • the resulting AMP can then be converted to ATP by an enzyme that catalyzes the reaction that produces ATP from AMP.
  • ATP is converted to AMP by luciferase and luminescence is produced and can be detected.
  • the sample may contain ATP and / or AMP.
  • the program means a cleaning program.
  • Cleaning programs include general hygiene management programs, SOPs, and SSOPs for managing the hygiene of food manufacturing and processing environments.
  • the cleaning program is sometimes referred to as a cleaning protocol.
  • the present invention provides a method for evaluating a cooking related utensil cleaning program.
  • This method includes the following steps: (i) cleaning cooking-related utensils according to the program; (ii) measuring the degree of contamination of the cleaned cooking-related utensils using the kit of the present invention; (iii) compare the degree of contamination measured in step (ii) with the references specified in the program; and (iv) If the measured degree of contamination exceeds the reference specified in the program, evaluate that the contamination in (i) does not remove the contamination sufficiently.
  • a cleaning method that is the same as or different from the cleaning method in the step (i) can be further performed.
  • the present invention provides a method for improving a cooking related utensil cleaning program.
  • This method includes the following steps: (i) cleaning cooking-related utensils according to the program; (ii) Use the kit of the present invention to measure the degree of contamination of the washed cooking-related utensils. (iii) Compare the pollution degree measured in step (ii) with the reference specified in the program. (iv) If the measured pollution degree exceeds the reference specified in the program, modify the cleaning process of i), and (v) Repeat steps (i) to (iv) until the pollution level is below the reference specified in the program.
  • a cooking program for cooking-related utensils can be created as step (o) before step (i).
  • cooking-related utensils refer to cooking utensils and the environment of the cooking site or those in the environment that may be contaminated.
  • cooking utensils for example, cutting boards, pans, frying pans, pressure cookers, iron plates, dishes, kitchen knives, chopsticks, chopsticks, spoons, forks, knives, etc., bowl characters, nets, colander, racks, cutting board stands, cooking utensils Examples include, but are not limited to, utensils for cooking such as containers for storing food, packaging containers, and packaging sheets, and utensils related thereto.
  • the environment at the cooking site refers to the environment at the cooking site, food processing plant, food provision facility, etc., which includes equipment, containers such as preparation tanks, piping, filling nozzles, belt conveyors, etc. in the food processing plant Examples include, but are not limited to, a part or place that touches a human hand such as a door knob, a handle of an appliance such as a refrigerator or an oven, a switch, or a receiver.
  • Contamination may include not only ATP, ADP, or AMP but also ATP-degrading enzyme and the like (for example, ATP-degrading enzyme, ADP-degrading enzyme, and enzymes that degrade ATP and ADP). Such contamination may cause ATP to be decomposed into ADP or ADP by the action of ATP degrading enzyme or the like even immediately after washing or during cleanliness measurement.
  • the kit and method of the present invention are also applicable to such contaminated cooking utensils.
  • Reference can be determined in advance based on standards required for food processing safety management.
  • “reference” refers to various indicators that can be associated with the degree of contamination or cleanliness.
  • the reference may be a threshold for a certain index that is found in the sample to be examined, with a threshold or higher being an indicator of contamination, while a value below the threshold is an absence of contamination or a clean state indicator.
  • An example of the indicator is the number of microorganisms.
  • the number of microorganisms is usually expressed as a number per mL (cells / mL).
  • the number of microorganisms can be evaluated by the number of colonies (cfu) formed when cultured under appropriate conditions per mL (cfu / mL).
  • the reference is a unit (cfu / mL) in which a colony of microorganisms is formed under a certain condition with respect to a test sample obtained by wiping off the tableware or cooking utensil. It can be expressed as a degree of contamination corresponding to. For example, if the acceptable cfu / mL is 1 cfu / mL, it can be judged that the sample in which 1 cfu / mL is detected is contaminated, and the sample below 1 cfu / mL can be judged to be clean. .
  • the reference for the contamination corresponding to 1 cfu / mL can be determined by examining the luminescence amount of the contaminated sample using the kit of the present invention.
  • the inspection standard of the coliform group inspection guideline for milk in Europe and the US is 5-10 cfu / mL
  • the inspection standard of Japanese Food Sanitation Inspection Law is 0.45 cfu / mL.
  • a facility's cleaning program related to milk production can make references based on these criteria.
  • the method of the present invention does not measure food itself having a specific cfu / mL standard, but measures the presence or absence of contamination after washing cooking-related utensils that handle food. For cooking-related utensils, if a level of contamination is detected, it is checked in advance whether it corresponds to a specific standard (for example, cfu / mL), and the degree of contamination corresponding to the standard is determined. Then, the contamination degree can be used as a threshold value as a reference.
  • the index for deriving the reference is not limited to cfu / mL, and may be various indices depending on the purpose.
  • Cleaning of cooking-related utensils includes washing with water, running water, standing washing, washing with detergent, ultrasonic washing, boil sterilization, heat sterilization, washing with washing utensils, washing with a dishwasher, drying with a dryer It may include steps such as drying, CIP (fixed cleaning) and COP (decomposition cleaning) in the production line of the food factory.
  • the detergent may be a soap, a synthetic detergent, or a surfactant, and examples of the surfactant include a cationic surfactant, an anionic surfactant, an amphoteric surfactant, and a nonionic surfactant.
  • a commonly used instrument such as a sponge, a brush, a scrubber, a cloth, or a paper towel can be used.
  • the sample may be a wipe of cooking-related utensils or water that has been washed with cooking-related utensils. Samples may be concentrated for evaluation testing.
  • the comparison between the measured contamination level and the reference may be performed manually or may be automated by the device.
  • the device may comprise a recording medium in which the reference is recorded in advance.
  • Improvement includes modifying the cleaning process.
  • modifying the cleaning process means changing the type of detergent used for cleaning, changing the concentration or amount of detergent used, changing the cleaning time or number of times, allowing the supervisor to supervise the cleaning operation, or Examples include, but are not limited to, strengthening supervision and ensuring that cleaning is performed as per the program.
  • the modification of the program may be repeated multiple times, and it can be repeated until the pollution level is below the reference specified in the program.
  • the correction of the program multiple times may be a correction for the same process or the same item, or may be a correction for a different process or a different item.
  • the cleaning program may include, for example, a step of washing or washing detergent-related utensils such as chopping boards and dishes used at the cooking site. At this time, if the washing is not sufficient, the cooking-related utensils remain contaminated.
  • the cleaning program it is possible to evaluate whether the cleaned cooking-related utensils are contaminated or whether the cleaning is sufficient, and the cleaning program can be modified appropriately.
  • the cleaning program may include, for example, a step of dipping a used dish or cooking utensil in a cleaning tank containing a detergent for a certain period of time at a cooking site. At this time, when the dirty dishes are added and immersed, the cleaning liquid in the cleaning tank is gradually contaminated. And if it exceeds a certain level, the tableware and cooking utensils taken out from the washing tank are not sufficiently washed, and contamination remains. By the method of the present invention, it can be evaluated how often the contamination can be prevented by replacing the cleaning liquid in the cleaning tank.
  • the method of the present invention can also be used for evaluating the cleaning power of a dishwasher. Put the contaminated dishes and the specified detergent into the dishwasher for cleaning. After washing, measure the degree of contamination of the dishes and compare with the reference. If the contamination exceeds the reference, the detergent used for the dishwasher (type, concentration) and operating conditions (temperature, time, water pressure, process repetition, etc.) may be modified.
  • the method of the present invention can also be used to evaluate the cleaning power of detergents. Wash with detergent and then measure the degree of contamination of the dishes and compare with the reference. If the contamination exceeds the reference, the usage of the detergent (concentration, amount, time, frequency, temperature, process repetition, etc.) may be modified.
  • the method of the present invention can be used not only to improve the cooking-related appliance cleaning program, but also to verify whether or not a correctly designed cleaning program is properly executed. For example, a facility where food poisoning has occurred can be subjected to a contamination test under cleaning conditions according to the program of the facility. If contamination is not detected by cleaning according to the program, it can be specified that there is no problem in the program itself, but there may be a problem in its execution. And the execution side can be improved. Such verification and improvement in execution are also widely included in the program improvement method of the present invention.
  • the kit of the invention comprises luciferase and luciferin.
  • metal ions such as magnesium, manganese, and calcium may also be included.
  • the necessary luciferase converts ATP, O 2 and luciferin to AMP, pyrophosphate, CO 2 and oxyluciferin, which results in luminescence.
  • the luciferase may be a natural luciferase or a genetically engineered recombinant luciferase variant.
  • the luciferase variant may be one that has been site-directed or randomly mutagenized. It may be a fusion protein with a protein having another function.
  • the luciferase mutant may have a desired property such as one having improved heat resistance and one having improved surfactant resistance.
  • the amount of luminescence of the luciferase can be determined by a suitable luminescence measuring device such as a luminometer (Berthold, Centro LB960 or Lumat 3 LB9508, Kikkoman Biochemifa, Lumitester C-110, Lumitester C-100, Lumitester PD-30.
  • a luminometer Boethold, Centro LB960 or Lumat 3 LB9508, Kikkoman Biochemifa, Lumitester C-110, Lumitester C-100, Lumitester PD-30.
  • the relative light emission intensity (RLU) obtained using a Lumitester PD-20 or the like can be evaluated as an index.
  • luminescence generated upon conversion from luciferin to oxyluciferin is measured.
  • a luminescence measuring device it is possible to measure with high sensitivity, and a device equipped with a photomultiplier tube (manufactured by 3M, etc.) or a device equipped with a photodiode (manufactured by Hygina, Neogen, etc.) may be used. it can.
  • the luciferase is not particularly limited as long as it uses ATP as a substrate, and those derived from bacteria, protozoa, animals, mollusks, and insects can be used.
  • Insects include beetle luciferases, such as the genus Photinus, such as the North American firefly (Photinus pyralis), the genus Photuris, such as the Photouris lucicrescens, the Shoturis pennsylvanica, the genus Luciola, such as the Luciola crucia , Firefly (Luciola lateralis), Japanese firefly (Luciola parvula), genus firefly (Pyrocoelia) genus, firefly (Lucidina biplagiata) firefly and Pyrophorus (Pyrophorus) genus beetle derived. Numerous luciferase genes have been reported, and their nucleotide sequences and amino acid sequences
  • the luciferase gene may be a wild type or may have a mutation.
  • the mutation may be a site-specific introduction or a random mutation.
  • Known mutations include those that improve the amount of luminescence as described in JP-A-2011-188787, mutations that increase the persistence of luminescence as described in JP-A-2000-197484, and Japanese Patent No. 2666561. Mutations that change the emission wavelength as described in JP-A No. 2003-512071, mutations that increase the resistance to surfactants as described in JP-A No. 11-239493, and International Publication No. 99/02697 Mutations that increase the substrate affinity as described in the pamphlet, JP 10-512750 A or JP 2001-518799 A, Japanese Patent No. 3048466, JP 2000-197487 A, JP 9-9 Mutations that increase stability, as described in JP 510610 and JP 2003-518912 Including but not limited to this.
  • the luciferase gene and its recombinant DNA can be prepared by conventional methods.
  • Japanese Patent Publication No. 7-112434 describes a Heike firefly luciferase gene.
  • JP-A-1-51086 describes a genji firefly luciferase gene.
  • the luciferase gene can be incorporated into a vector such as a plasmid, bacteriophage, cosmid, etc., to transform or transduce an appropriate host.
  • the host can be a microorganism, a bacterium such as E. coli, or a yeast.
  • the transformed host capable of producing luciferase can be cultured by various known methods.
  • tryptone, yeast extract, meat extract, peptone, corn steep liquor, or one or more nitrogen sources such as soybean or wheat bran leachate, sodium chloride, monopotassium phosphate, dipotassium phosphate, chloride
  • nitrogen sources such as soybean or wheat bran leachate, sodium chloride, monopotassium phosphate, dipotassium phosphate, chloride
  • inorganic salts such as magnesium, ferric chloride, magnesium sulfate, or manganese sulfate are added, and if necessary, saccharide raw materials, vitamins, and the like are added.
  • the initial pH of the medium can be 7-9, for example.
  • the culture can be performed, for example, at 30 to 40 ° C. for 2 to 24 hours by aeration and agitation culture, shaking culture, stationary culture, or the like. After the culture, luciferase is recovered from the culture by a known method.
  • the cells are subjected to ultrasonic crushing treatment, grinding treatment or the like by a conventional method, or luciferase is extracted using a lytic enzyme such as lysozyme.
  • a lytic enzyme such as lysozyme.
  • the obtained extract is filtered, centrifuged, etc., nucleic acid is removed with streptomycin sulfate, if necessary, and ammonium sulfate, alcohol, acetone, etc. are added thereto and fractionated to obtain a crude enzyme.
  • the crude enzyme may be further purified by various gel filtration and chromatographic techniques.
  • a commercially available luciferase can also be used, for example, a luciferase of Kikkoman Biochemifa, catalog number 61314 can be used. This luciferase is described in Japanese Patent Application Laid-Open No. 11-239493 (Patent No. 3794628) (SEQ ID NO: 1 in this document).
  • commercially available luciferases of molecular probes (registered trademark) from Sigma-Aldrich, Promega, and Life Technology can be used.
  • the luciferin may be any luciferin that is recognized as a substrate by the luciferase used, and may be natural or chemically synthesized. Any known luciferin derivative can also be used.
  • the basic skeleton of luciferin is imidazopyrazinone, and there are many tautomers. Examples of luciferin include firefly luciferin. Firefly luciferin is a substrate for firefly luciferase (EC 1.13.12.7). Luciferin derivatives can be those described in JP-A-2007-91695, JP-T 2010-523149 (International Publication No. 2008/127777) and the like.
  • the final concentration in the luciferase measurement system is 0.001 ⁇ g protein / mL or more, 0.01 ⁇ g protein / mL or more, 0.02 ⁇ g protein / mL or more when the absorbance at 280 nm is luciferase concentration (mg protein / mL), 0.05 ⁇ g protein / mL or more, 0.10 ⁇ g protein / mL or more, 0.20 ⁇ g protein / mL or more, or 0.25 ⁇ g protein / mL or more.
  • the final concentration in the luciferase measurement system is 1 ⁇ g protein / mL or less, 0.5 ⁇ g protein / mL or less, or 0.3 ⁇ g protein / mL or less when the absorbance at 280 nm is luciferase concentration (mg protein / mL). be able to.
  • the final concentration of the luciferin or luciferin derivative in the measurement system may be 0.01 mM to 20 mM, 0.05 mM to 20 mM, 0.1 mM to 20 mM, 0.5 mM to 10 mM, such as 0.75 mM to 5 mM.
  • Enzymes that catalyze reactions that produce ATP from ADP employ enzymes that catalyze reactions that produce ATP from ADP.
  • ADP present in the system is converted to ATP by an enzyme that catalyzes a reaction that generates ATP from ADP.
  • ATP is converted to AMP by luciferase and light is emitted.
  • any known enzyme can be used, for example, a kinase having ATP generating ability.
  • the kinase having ATP generation ability include pyruvate kinase, acetate kinase, creatine kinase, polyphosphate kinase, hexokinase, glucokinase, glycerol kinase, fructokinase, phosphofructokinase, riboflavin kinase, fructose bisphosphatase and combinations thereof However, it is not limited to this.
  • Pyruvate kinase (EC 2.7.1.40) converts phosphoenolpyruvate to pyruvate in the glycolytic system, where ADP is converted to ATP.
  • This reaction is an Ergon reaction with negative Gibbs energy and is irreversible under natural conditions: PEP + ADP ⁇ Pyruvate + ATP
  • the reverse reaction is catalyzed by pyruvate carboxylase and phosphoenolpyruvate carboxykinase in gluconeogenesis to produce PEP and ADP from ATP and pyruvate.
  • various enzymes are mixed in the system, and the above reaction can proceed in both directions.
  • ADP can be converted to ATP. Further, if not only phosphoenolpyruvate but also pyruvate kinase is present in the system, it is considered that ADP is more converted to ATP.
  • animals derived from microorganisms such as animals, such as a rabbit, a rat, a chicken, yeast, Bacillus stearothermophilus (Bacillus stearothermophilus), can be used.
  • Acetate kinase (AK) Acetate kinase (EC 2.7.2.1) catalyzes the conversion between ATP and acetic acid and ADP and acetylated phosphate in the presence of cations: ATP + acetic acid ⁇ ⁇ ADP + acetylated phosphate
  • Acetate kinase (AK) is also called ATP: acetate phosphotransferase, acetyl kinase. As used herein, these terms can be interchanged.
  • ATP and acetic acid are used to generate ADP and acetylated phosphate, and ultimately promote the reaction to produce acetyl CoA.
  • the system contains acetylated phosphate and ADP generated from acetyl-CoA, it can be converted to acetic acid and ATP.
  • Escherichia coli Escherichia coli
  • Bacillus stearothermophilus Bacillus stearothermophilus
  • Costridium pasteurianum Costridium pasteurianum
  • Lactobacillus delbrucki Lactobacillus delbruckii
  • the one derived from Veillonella alcalescence can be used.
  • Creatine kinase (CK) Creatine kinase (EC 2.7.3.2) mediates the conversion reaction between creatine and ATP and creatine phosphate and ADP: Creatine + ATP ⁇ ⁇ Creatine phosphate + ADP Creatine kinase (CK) is also called creatine phosphokinase (CPK) or phosphocreatine kinase. As used herein, these terms can be interchanged. Usually, creatine phosphate and ADP are produced from creatine and ATP in animal muscles and the like.
  • this reaction is a reversible reaction, and if creatine phosphate and ADP are present in a high concentration in the system, the reaction proceeds in the reverse direction, and creatine and ATP can be generated.
  • cytoplasmic creatine kinase is composed of two subunits B or M. Therefore, three isozymes, CK-MM, CK-BB and CK-MB may exist depending on the combination of subunits.
  • the isozyme pattern varies depending on the tissue, but any combination can be used in the present invention.
  • the thing derived from an animal can be used, For example, the thing derived from a rabbit, a chicken, a cow, a pig, a carp, a catfish, a frog is mentioned.
  • Polyphosphate kinase (PPK) Polyphosphate kinase (EC 2.7.4.1) catalyzes the reaction of converting polyphosphate (PolyP n ) and ADP to polyphosphate (PolyP n-1 ) and ATP: ADP + PolyP n ⁇ ⁇ ATP + PolyP n-1 Polyphosphate kinase (PPK) is also called ATP: polyphosphate phosphotransferase. As used herein, these terms can be interchanged. PPK is involved in oxidative phosphorylation in vivo. If polyphosphoric acid (n) and ADP are present in the system, they can be converted to polyphosphoric acid (n-1) and ATP.
  • microorganism-derived things such as Escherichia coli (Escherichia coli), yeast, Corynebacterium xerosis (Corynebacterium xerosis), can be used.
  • Riboflavin kinase (EC 2.7.1.26), also described as FMNK, catalyzes the reaction of converting riboflavin and ATP to riboflavin phosphate (FMN) and ADP: ATP + riboflavin ⁇ ⁇ ADP + FMN Riboflavin kinase belongs to ATP: riboflavin 5′-phosphotransferase (also referred to as flavokinase).
  • ATP riboflavin 5′-phosphotransferase
  • flavokinase also referred to as flavokinase.
  • those derived from microorganisms and animals can be used, and examples include those derived from yeast, rat, and legume (Phaseolus radiatus).
  • Phosphofructokinase 1 (EC 2.7.1.11), also described as PFK1, converts fructose-6-phosphate (Fru6P) and ATP to fructose-1,6-bisphosphate (Fru1,6-BP) and ADP Catalyze the reaction to: Fru6P + ATP ⁇ ⁇ Fru1,6-BP + ADP Phosphofructokinase 1 belongs to phosphofructokinase. In the present specification, phosphofructokinase 1 is sometimes referred to as Fru-1,6BPK.
  • those derived from animals and microorganisms can be used, for example, those derived from microorganisms include baker's yeast, beer yeast, Clostridium pasteurianum, Escherichia coli, Bacillus Examples are those derived from Bacillus licheniformis.
  • Fructose bisphosphatase Fructose bisphosphatase (EC 3.1.3.11), also described as FBPase, is a reaction that converts fructose-1,6-bisphosphate (Fru1,6-BP) and ADP to fructose-6-phosphate (Fru6P) and ATP. Catalyze: Fru1,6-BP + ADP ⁇ ⁇ Fru6P + ATP Fructose bisphosphatase may be described as FBP or FBP1. Although not particularly limited, those derived from animals, plants, and microorganisms can be used, and examples include those derived from rabbits and chickens.
  • Pyruvate-phosphate dikinase (EC 2.7.9.1) is a reaction between ATP, pyruvate and orthophosphate and adenosine monophosphate (AMP), phosphoenolpyruvate (PEP) and pyrophosphate (PPi).
  • AMP adenosine monophosphate
  • PEP phosphoenolpyruvate
  • PPi pyrophosphate
  • PPDK Pyruvate-phosphate dikinase
  • ATP pyruvate, phosphate phosphotransferase, pyruvate orthophosphate dikinase, pyruvate phosphate ligase.
  • PPDK Pyruvate-phosphate dikinase
  • PPDK usually converts pyruvic acid to PEP, and in the process, one ATP molecule is consumed and converted to AMP. The reaction is divided into the following three reversible reactions. 1. The enzyme PPDK binds to ATP and results in conversion to AMP and diphosphorylated PPDK. 2.
  • Diphosphorylated PPDK binds to inorganic phosphoric acid, resulting in diphosphoric acid and monophosphorylated PPDK. 3. Monophosphorylated PPDK binds to pyruvic acid, yielding PEP and PPDK again. At this time, if the PEP concentration present in the system is high, the reaction proceeds in the reverse direction as follows.
  • PEP binds to PPDK, producing monophosphorylated PPDK and pyruvate.
  • Diphosphorylated PPDK and inorganic phosphoric acid are produced from diphosphoric acid and monophosphorylated PPDK.
  • PPDK and ATP are produced from biphosphorylated PPDK and AMP.
  • the PPDK is not particularly limited, and examples thereof include, for example, Microbispora thermorosea, Propionibacterium shremanii, Bacteroides symbiosus, Entamoeba histolytica, Acetobacter xylinum described in JP-A-8-168375 (Patent No. 3181801). And those derived from microorganisms such as Propionibacter shermanii, and those derived from plants such as corn and sugarcane.
  • ADK Addenylate kinase
  • Adenylate kinase (EC 2.7.4.3), also called adenylate kinase, catalyzes the following reaction in the presence of metal ions: ATP + AMP ⁇ ⁇ 2ADP This reaction is reversible.
  • ADK is an example of an enzyme that catalyzes a reaction that generates ADP from AMP. When ADK is combined with PK or the like, ADP is converted to ATP, and as a result, ATP, ADP, and AMP can be measured.
  • ADK is not particularly limited, and examples thereof include those derived from microorganisms such as yeast and those derived from animals such as rabbits, pigs, cows, rats, and pigs.
  • Pyruvate water dikinase (EC 2.7.9.2) catalyzes the following reaction: ATP + pyruvate + H 2 O ⁇ ⁇ AMP + phosphoenolpyruvate (PEP) + phosphate (P) Pyruvate water dikinase is also called phosphoenolpyruvate synthase; pyruvate water dikinase (phosphorylation); PEP synthetase; phosphoenolpyruvate synthetase; phosphoenolpyruvic synthetase; phosphopyruvate synthetase. As used herein, these terms can be interchanged.
  • ⁇ ATP generation from AMP and PEP can be promoted by using PWDK together with PEP.
  • PWDK When PWDK is combined with PK or the like, ADP is converted to ATP, and as a result, ATP, ADP, and AMP can be measured.
  • the PWDK is not particularly limited.
  • the kit of the present invention may comprise an RNase.
  • the method of the present invention may use an RNase.
  • the RNase means an RNase that is not derived from a sample.
  • RNase means an enzyme that catalyzes a reaction that generates 5′-mononucleotides (AMP, GMP, CMP, and UMP) from RNA, and examples thereof include the following: (1) Endonuclease S1 (EC3.1.30.1), (2) Venom exonuclease (EC3.1.15.1), (3) Phosphodiesterase One ( Phospho diesterase 1) (EC3.1.4.1).
  • the endonuclease s-one includes nuclease pi-one (Nuclease P1), mung bean nuclease (Mung beans nuclease), and neurospora crassa nuclease.
  • the kit of the invention does not contain RNase or does not contain a substantial amount of RNase.
  • the methods of the invention do not use RNases or do not use substantial amounts of RNases.
  • RNase derived from the sample may be included in the reaction system.
  • the “substantial amount of RNase” refers to the effect of the kit or method of the present invention (for example, the effect of providing an accurate contamination detection method that is hardly affected by the ATP degradation activity). An amount of RNase that has no effect.
  • the final concentration in the reaction system is 0.3 U / mL or less, 0.15 U / mL or less, 0.1 U / mL or less, 0.05 U / mL or less, 0.01 U / mL as an example not including the substantial amount of RNase
  • a kit containing an RNase of 0.001 U / mL or less, or a method using such an amount of RNase is mentioned.
  • the enzyme unit of RNase focuses on the RNA resolution of the enzyme, and the activity unit (U) of the enzyme having RNA resolution is 37 ° C., and 1.0 ⁇ mol of substrate per minute is acid-soluble. Defined as the amount of enzyme converted to nucleotides.
  • the enzyme unit of Nuclease P1 is defined as the amount of enzyme that converts 1.0 ⁇ mol of substrate into acid-soluble nucleotides per minute at 37 ° C., pH 5.3 (as defined in the definition of the enzyme activity of Nuclease P1).
  • the Merck catalog http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/General_Information/nuclease_p1.pdf).
  • the kit or method of the invention comprises, uses, or substantially comprises or uses an RNase
  • the RNase may or may not contribute to the luminescence reaction by luciferase. You don't have to contribute.
  • the kit of the present invention substantially contains an RNase
  • the kit of the present invention may or may not contain an enzyme that generates ATP from AMP.
  • the methods of the invention may or may not use enzymes that produce ATP from AMP.
  • the present invention provides for the ATP, ADP, And the amount of luminescence is measured before it affects the measurement of AMP.
  • the amount of luminescence when RNase is included is 2 times or less, 1.8 times or less, 1.5 times or less, 1.2 times or less, 1.1 times or less, or equivalent to the amount of luminescence without RNase. Measurements can be made at such times.
  • the measurement time can be appropriately set according to the amount of RNase, for example, within 10 minutes, within 5 minutes, within 4 minutes, preferably within 3 minutes, within 2 minutes, or within 1 minute, within 30 seconds, or Can be within 10 seconds. Even if a large amount of RNase is contained, the action of RNase can be reduced by shortening the reaction time.
  • the present invention may be used on samples that are free of RNA or substantially free of RNA.
  • the amount of luminescence when RNase is included is 2 times or less, 1.8 times or less, 1.5 times or less, 1.2 times or less, 1.1 times or less than the amount of luminescence when RNAase is not included, Or the sample which becomes equivalent is mentioned.
  • the present invention provides an enzyme that generates AMP from ADP, an enzyme that generates ATP from AMP (eg, PPDK), a kit including luciferin, luciferase, and a metal salt, and a measurement method using the kit.
  • an enzyme that generates AMP from ADP eg, PPDK
  • an enzyme that generates ATP from AMP eg PPDK
  • ADP is converted to AMP
  • AMP is converted to ATP.
  • ATP, ADP, and AMP should be measured.
  • the kit can further include PEP and PPi.
  • the enzyme that generates ATP from AMP is as described above, and examples of the enzyme that generates AMP from ADP include ADP-dependent hexokinase and apyrase.
  • ADP-dependent hexokinase (EC 2.7.1.147), also called ADP-specific hexokinase, catalyzes the following reaction: D-glucose + ADP ⁇ ⁇ D-glucose-6-phosphate + AMP
  • [Apyrase] Apyrase (EC 3.6.1.5), also called adenosine diphosphatase, ADPase, ATP diphosphatase, or ATP diphosphohydrolase, catalyzes the following two reactions: ATP + H 2 O ⁇ ⁇ ADP + phosphoric acid (P) ADP + H 2 O ⁇ ⁇ AMP + phosphoric acid (P)
  • the enzymes that catalyze the reaction of generating ATP from ADP, PPDK, PWDK, and ADK, and the enzyme that generates AMP from ADP may be collectively referred to as an enzyme having ATP generating ability.
  • any known ones such as those derived from microorganisms, bacteria, eukaryotes, protists, plants, animals, etc. can be used, for example, commercially available ones. Can be used.
  • the addition amount of the enzyme can be appropriately set according to the target concentration and reaction system.
  • the enzyme having ATP generation ability is added so that the activity unit in the measurement system is 0.001 U or more, 0.01 U or more, 0.1 U or more, 1 U or more, 2 U or more, 3 U or more, 4 U or more, or 5 U or more. be able to.
  • the enzyme having ATP generating ability is added so that the activity unit in the measurement system is 10,000 U or less, 1000 U or less, 100 U or less, 50 U or less, 10 U or less, 9 U or less, 8 U or less, 7 U or less, or 6 U or less. be able to. A person skilled in the art can appropriately determine the amount of the enzyme added.
  • a substrate for each enzyme can be added, and is not particularly limited.
  • phosphoenolpyruvate and pyrophosphate for PPDK For PK, AK, CK, PPK, FMNK, PFK1, and FBPase, use phosphoenolpyruvate, acetyl phosphate, creatine phosphate, polyphosphate, riboflavin phosphate, fructose-1,6-bisphosphate, respectively. it can.
  • PWDK phosphoenolpyruvate and phosphate can be used.
  • glucose can be used for ADP-dependent hexokinase.
  • the kits of the invention further comprise these substrates. In certain embodiments, the methods of the invention may further use these substrates.
  • PEP phosphoenolpyruvate
  • the method of the present invention may use phosphoenolpyruvate (PEP).
  • PEP phosphoenolpyruvate
  • the concentration of PEP to be used includes 0.001 mM to 4500 mM, for example, 2.1 mM as the final concentration.
  • the method of the present invention may use pyrophosphate (PPi).
  • PPi pyrophosphate
  • concentration of PPi used is 0.001 mM to 2000 mM, for example, 0.2 mM as the final concentration.
  • the reaction reagent can also include an enzyme stabilizer such as bovine serum albumin or gelatin that protects reporter molecules such as luciferase from degradation.
  • the reaction reagent may also be added with a substance that improves pH adjustment and storage stability.
  • suitable pH buffer HPES, Tricine, Tris, phosphate buffer, acetate buffer, etc.
  • reducing agent dithiothreitol (DTT), 2-mercaptoethanol, etc.
  • sugar glucose, sucrose, trehalose, etc.
  • the kit of the invention may include instructions for use.
  • the instructions for use may be one in which the procedure of the cleaning program evaluation method of the present invention, the procedure of the cleaning program improvement method and the method of using the kit of the present invention are described.
  • ATP assay method using luciferase The assay method using luciferase is described below. The conditions are exemplary. Prepare an ATP measurement reagent containing: MES 1 mM Magnesium acetate 5.1 mM Potassium pyrophosphate 0.15mM Potassium phosphoenolpyruvate 2.1 mM Luciferin 0.8 mM Tricine 25mM Luciferase 12.5 ⁇ g protein / mL (The absorbance at 280nm is the luciferase concentration (mg protein / mL).) Add 0.1 mL of sample solution containing ATP to 0.1 mL of the above ATP measurement reagent and measure luminescence.
  • the amount of luminescence can be measured using a known luminometer (Berthold Centro LB960 or Lumat3 LB9508, Kikkoman Biochemifa luminometer, etc.).
  • Luminescence can be described as a relative luminescence unit (RLU) relative to a certain standard.
  • RLU relative luminescence unit
  • ATP + ADP assay method 1 (luciferase + PK)
  • the ATP + ADP assay method is described below.
  • the conditions are exemplary.
  • ATP + ADP assay method 2 (luciferase + AK)
  • the ATP + ADP assay method is described below. The conditions are exemplary.
  • ATP + AMP assay method (luciferase + PPDK)
  • the ATP + AMP assay method will be described below.
  • the conditions are exemplary.
  • a reagent for measuring ATP + AMP containing the following is prepared.
  • ATP + AMP + ADP assay method 1 (luciferase + PK + PPDK)]
  • the ATP + AMP + ADP assay method will be described below.
  • the conditions are exemplary.
  • ATP + ADP + AMP assay method 2 (luciferase + PK + ADK)] The ATP + AMP + ADP assay method will be described below. The conditions are exemplary.
  • Prepare ATP and AMP + ADP measurement reagents including the following: Magnesium acetate 5.1 mM Potassium pyrophosphate 0.15mM Potassium phosphoenolpyruvate 2.1 mM Luciferin 0.8 mM Tricine 25mM Luciferase 12.5 ⁇ g protein / mL (The absorbance at 280nm is the luciferase concentration (mg protein / mL).) PK 25U / mL ADK 500U / mL
  • ATP + ADP + AMP assay method 3 (luciferase + PPDK + ADP-dependent hexokinase or apyrase)]
  • the ATP + AMP + ADP assay method will be described below. The conditions are exemplary.
  • ADP measurement reagents including the following: MES 1 mM Magnesium acetate 5.1 mM Potassium pyrophosphate 0.15 mM Potassium phosphoenolpyruvate 2.1 mM Luciferin 0.8 mM Tricine 25 mM Luciferase 12.5 ⁇ g protein / mL (The absorbance at 280 nm is the luciferase concentration (mg protein / mL).) ADP-dependent hexokinase 30 U / mL + glucose 10 mM or apyrase 1 U / mL PPDK 2U / mL
  • ATP (conventional 1 component) includes Lucipak II (manufactured by Kikkoman Biochemifa), ATP + AMP (conventional 2 components) includes Lucipak Pen (manufactured by Kikkoman Biochemifa), ATP + ADP (present invention 2 components) Is a reagent in which 0.1 mg of phosphoenolpyruvate (PEP) and 5 U of pyruvate kinase (Lee Biosolutions Inc. Catalog No.
  • Lucipak II 500-20 are added to Lucipak II, and Lucipak is added to ATP + ADP + AMP (3 components of the present invention).
  • ATP conventional one component
  • ATP + ADP two components of the present invention
  • Lumitester C-110 Lumitester C-110 (Kikkoman Biochemifa).
  • Luminity tester PD-30 manufactured by Kikkoman Biochemifa was used to measure the amount of luminescence for (conventional two components) and ATP + ADP + AMP (three components of the present invention). In this reagent, when the same concentration of ATP solution is measured, the same amount of luminescence is displayed.
  • Lucipak II and Lucipak Pen can be replaced with a composition comprising: Luciferase manufactured by Kikkoman Biochemifa (catalog number: 61314) was used, and PPDK described in JP-A-8-168375 (Japanese Patent No. 3181801) was used.
  • ⁇ Lushi Pack II composition Magnesium acetate 7mM Luciferin 0.5 mM Tricine 25mM Luciferase 0.7 ⁇ g protein / mL (The absorbance at 280 nm is the luciferase concentration (mg protein / mL).) ⁇ Lusipak Pen composition> Magnesium acetate 7mM Luciferin 0.5 mM Tricine 25mM Luciferase 0.2 mg protein / mL (The absorbance at 280 nm is the luciferase concentration (mg protein / mL).) Potassium pyrophosphate 0.2mM Potassium phosphoenolpyruvate 1.4 mM PPDK 1.3U / mL
  • ATP (conventional 1 component), ATP + AMP (conventional 2 component), ATP + ADP + AMP (3 components of the present invention) and ATP + ADP (2 components of the present invention) were measured: Washing conditions No sticking of meat, no cleaning process (negative control) 2. Stick meat and do not wash (positive control) 3. Let the meat adhere, and then run for about 10 seconds with running water (washing with running water) 4). Adhere meat, then rub lightly with a sponge and wash with running water for about 10 seconds (sponge + washing with running water) 5).
  • the results are as follows.
  • the luminescence unit is RLU and the bacterial count unit is cfu / mL.
  • the cleanliness evaluation results by the cleaning method were divided according to the number and types of components to be measured.
  • Luminescence was lower than in the case of washing with sponge + running water. This tendency was not consistent with the case where two components of ATP + AMP were measured. Therefore, if only ATP is measured, the cleanliness evaluation may not be stable.
  • the total luminescence when ATP + AMP was measured was lower than that of ATP + ADP.
  • the total amount of luminescence in the three-component measurement was the largest, but when ATP + ADP was measured, the amount of luminescence was the same. If two components of ATP + ADP were measured, the three components of ATP + ADP + AMP were It was shown that almost the same evaluation can be performed as when measured.
  • This test is a kind of accelerated test using conditions suitable for the growth of bacteria, but it is considered that washing is performed to reduce the number of growing bacteria at actual cooking sites. In that case, contamination cannot be detected by the amount of luminescence when ATP + AMP is measured, but it may be detected by measuring ATP + ADP or ATP + ADP + AMP. Similarly, for samples containing ATP-degrading enzymes, etc., the amount of luminescence when ATP + AMP is measured cannot be detected, but it may be detected by measuring ATP + ADP or ATP + ADP + AMP. It is done.
  • Example 2 Construction of an ATP + ADP + AMP measurement system using an enzyme that catalyzes the reaction that generates AMP from ADP.
  • ADP-dependent hexokinase (ADP + AMP) is an enzyme that catalyzes the reaction that generates AMP from ADP.
  • Asahi Kasei Pharma, T-93 ADP-HKTII and glucose were added to investigate whether ATP + ADP + AMP could be measured.
  • the composition of the luminescent reagent is as follows.
  • apyrase (Sigma A6536), an enzyme that catalyzes the reaction to generate AMP from ADP, is added to the luminescent reagent for ATP + AMP measurement of the same composition instead of hexokinase, and ATP + ADP + AMP It was investigated whether measurement was possible.
  • the composition of the luminescent reagent is as follows.
  • a calibration curve was prepared by calculating the molar amounts of ATP, ADP, and AMP in the solution during luminescence. The results are shown in FIGS. 4-1 to 4-3. These results indicate that ATP + ADP + AMP can be measured by using PPDK that catalyzes the reaction that generates ATP from AMP and an enzyme that catalyzes the reaction that generates AMP from ADP. Yes. Using these methods, the degree of contamination of the cooking-related utensils can be evaluated.
  • the degree of contamination of cooking-related utensils can be measured.
  • the cleaning method and cleaning program for cooking-related utensils can be modified or altered according to the degree of contamination.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Zoology (AREA)
  • Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Microbiology (AREA)
  • General Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Genetics & Genomics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Plasma & Fusion (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Cette invention concerne un procédé permettant d'évaluer ou d'améliorer un programme de nettoyage d'instruments de cuisson, où ledit procédé d'évaluation et procédé d'amélioration de programme de nettoyage utilisent un kit de mesure d'ATP et d'ADP. Le procédé comprend : une étape (i) où les instruments de cuisson sont nettoyés selon le programme ; une étape (ii) où le kit est utilisé et le degré de contamination est mesuré sur les instruments de cuisson nettoyés ; une étape (iii) où le degré de contamination mesuré à l'étape (ii) est comparé à une référence prescrite par le programme ; une étape (iv) où une évaluation est faite selon laquelle la contamination n'a pas été correctement éliminée lors de l'étape de nettoyage (i) ou selon laquelle l'étape de nettoyage (i) est corrigée, si le degré de contamination mesuré dépasse la référence prescrite par le programme ; et une étape (v) où les étapes (i) à (iv) sont répétées jusqu'à ce que le degré de contamination ne soit plus supérieur à la référence prescrite par le programme. Le kit utilisé peut comprendre : une enzyme qui catalyse une réaction qui génère de l'ATP à partir d'ADP ; de la luciférine ; de la luciférase ; et des sels métalliques. Le kit utilisé peut également comprendre : une enzyme qui catalyse une réaction qui génère de l'ATP à partir d'AMP ; une enzyme qui catalyse une réaction qui génère de l'AMP à partir d'ADP ; de la luciférine ; de la luciférase ; et des sels métalliques.
PCT/JP2018/004722 2017-02-09 2018-02-09 Kit permettant de mesurer le degré de contamination d'instruments de cuisson et procédé d'évaluation d'un programme de nettoyage d'instruments de cuisson Ceased WO2018147443A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2018567526A JP7227008B2 (ja) 2017-03-24 2018-02-09 調理関連器具汚染度測定用キット及び調理関連器具洗浄プログラム評価方法

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
JP2017-021952 2017-02-09
JP2017021952 2017-02-09
JP2017-058940 2017-03-24
JP2017058940 2017-03-24
JP2017192606 2017-10-02
JP2017-192606 2017-10-02

Publications (1)

Publication Number Publication Date
WO2018147443A1 true WO2018147443A1 (fr) 2018-08-16

Family

ID=63107206

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2018/004722 Ceased WO2018147443A1 (fr) 2017-02-09 2018-02-09 Kit permettant de mesurer le degré de contamination d'instruments de cuisson et procédé d'évaluation d'un programme de nettoyage d'instruments de cuisson

Country Status (1)

Country Link
WO (1) WO2018147443A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112274079A (zh) * 2020-09-25 2021-01-29 华帝股份有限公司 用于烹饪设备的清洁控制方法及应用其的烹饪设备
JP2021056195A (ja) * 2019-09-28 2021-04-08 佛山克情日用品有限公司 食器清浄度の測定設備
WO2021162123A1 (fr) 2020-02-14 2021-08-19 キッコーマン株式会社 Composition liquide pour mesurer l'atp, et amp et/ou adp dans des échantillons
CN114405937A (zh) * 2022-01-17 2022-04-29 内蒙古蒙牛乳业（集团）股份有限公司 一种乳制品生产设备的清洗方法、清洗装置和电子设备

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH10262697A (ja) * 1997-03-27 1998-10-06 Asahi Chem Ind Co Ltd Ａｄｐの測定法
JPH1169997A (ja) * 1997-08-28 1999-03-16 Kikkoman Corp 清浄度検査試薬およびその試薬を用いる清浄度検査法
JP2003035673A (ja) * 2001-07-23 2003-02-07 Kikkoman Corp 清浄度測定装置および清浄度改善システム

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH10262697A (ja) * 1997-03-27 1998-10-06 Asahi Chem Ind Co Ltd Ａｄｐの測定法
JPH1169997A (ja) * 1997-08-28 1999-03-16 Kikkoman Corp 清浄度検査試薬およびその試薬を用いる清浄度検査法
JP2003035673A (ja) * 2001-07-23 2003-02-07 Kikkoman Corp 清浄度測定装置および清浄度改善システム

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
"Enabling simultaneous inspection of ATP+ADP+AMP by ATP wiping inspection", JAPAN FOOD SCIENCE, vol. 56, 5 April 2017 (2017-04-05), pages 49 *
"From ATP wiping inspection to A3 (ATP+ADP+AMP) - Pace of 20 years and future development of Lumi-tester and lucipac", HAZARD ANALYSIS AND CRITICAL CONTROL POINT (HACCP), vol. 23, 1 June 2017 (2017-06-01), pages 100 - 108 *
ANONYMOUS: "Effective utilization example ofATP wiping inspection in establishing Food Hygiene 7S", KIKKOMAN CORP, 24 August 2016 (2016-08-24), pages 1 - 7, XP055537157, Retrieved from the Internet <URL:http://biochemifa.kikkoman.co.jp/pdf/j/j_jirei/pf_029_1y160801.pdf> [retrieved on 20171130] *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2021056195A (ja) * 2019-09-28 2021-04-08 佛山克情日用品有限公司 食器清浄度の測定設備
WO2021162123A1 (fr) 2020-02-14 2021-08-19 キッコーマン株式会社 Composition liquide pour mesurer l'atp, et amp et/ou adp dans des échantillons
JPWO2021162123A1 (fr) * 2020-02-14 2021-08-19
CN112274079A (zh) * 2020-09-25 2021-01-29 华帝股份有限公司 用于烹饪设备的清洁控制方法及应用其的烹饪设备
CN114405937A (zh) * 2022-01-17 2022-04-29 内蒙古蒙牛乳业（集团）股份有限公司 一种乳制品生产设备的清洗方法、清洗装置和电子设备
CN114405937B (zh) * 2022-01-17 2023-10-10 内蒙古蒙牛乳业（集团）股份有限公司 一种乳制品生产设备的清洗方法、清洗装置和电子设备

Similar Documents

Publication Publication Date Title
WO2018147443A1 (fr) Kit permettant de mesurer le degré de contamination d&#39;instruments de cuisson et procédé d&#39;évaluation d&#39;un programme de nettoyage d&#39;instruments de cuisson
Norwood et al. Adherence of Listeria monocytogenes strains to stainless steel coupons
US9102976B2 (en) Biological indicator
CN101578374B (zh) 用于检测样品中活微生物细胞的分析方法
JPH09182600A (ja) Ａｔｐ消去剤、ａｔｐ消去法、それを用いた生物細胞測定試薬及び生物細胞測定法
Sakakibara et al. Enumeration of bacterial cell numbers by amplified firefly bioluminescence without cultivation
JP7328763B2 (ja) 生体関連サンプル及び生体関連器具の清浄度測定キット及び方法
US8003341B2 (en) Method of amplifying ATP and use thereof
Manios et al. Control of Listeria monocytogenes in the processing environment by understanding biofilm formation and resistance to sanitizers
JP7227008B2 (ja) 調理関連器具汚染度測定用キット及び調理関連器具洗浄プログラム評価方法
JP2018126128A (ja) 調理関連器具汚染度測定用キット及び調理関連器具洗浄プログラム評価方法
US20230085844A1 (en) Liquid composition for measuring atp, and amp and/or adp in a sample
JPH1169997A (ja) 清浄度検査試薬およびその試薬を用いる清浄度検査法
JP2018126127A (ja) 生体関連サンプル及び生体関連器具の清浄度測定キット及び方法
Hansen et al. ATP bioluminescence–for kitchen hygiene and cleaning control of surgical instruments
EP4560025A1 (fr) Procédé de détection de micro-organismes ou de cellules, ou de substances apparentées à des micro-organismes ou à des cellules, et kit associé
WO2024019166A1 (fr) Procédé de détection de micro-organismes ou de cellules, ou de substances apparentées à des micro-organismes ou à des cellules, dans une solution, ainsi que kit associé
SUZUKI et al. A novel bioluminescent cycling assay for ATP and AMP using pyruvate orthophosphate dikinase
Motlhatlhedi Development of a methodology for establishing mixed culture biofilms and determination of their ATP content: A laboratory study
JP4492915B2 (ja) 食品の状態のモニター法
Eden et al. Microbiological growth-based and luminescence methods of food analysis
BG65685B1 (bg) Метод за установяване на наличие на атибиотици и инхибиторни вещества в млякото

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18751084

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2018567526

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 18751084

Country of ref document: EP

Kind code of ref document: A1