CN109757645A - The method for inducing gemma release DPA to reduce heat resistance - Google Patents
The method for inducing gemma release DPA to reduce heat resistance Download PDFInfo
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- CN109757645A CN109757645A CN201811531062.XA CN201811531062A CN109757645A CN 109757645 A CN109757645 A CN 109757645A CN 201811531062 A CN201811531062 A CN 201811531062A CN 109757645 A CN109757645 A CN 109757645A
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- gemma
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- heat resistance
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Abstract
The invention proposes a kind of methods that induction gemma release DPA reduces heat resistance, and at room temperature by product subject to sterilization, in the moulding pressure of 350~450kPa, carrying out amplitude is 100%, and frequency is 25~35min of ultrasonic treatment of 18~22khz.With the 1 × 10 of 30mL7.3The spore solution of CFU/mL is as product subject to sterilization, after being handled according to method proposed by the invention, can discharge 92.54% DPA, so that its heat resistance be greatly lowered.The present invention discharges DPA using method induction gemma associated with ultrasonic wave and high pressure；Compared with individual HIGH PRESSURE TREATMENT, ultrasonic treatment, acoustic pressure Combined Treatment can induce gemma to discharge most of DPA release, reduce gemma resistance, while ultrasonic combined HIGH PRESSURE TREATMENT can reduce cost compared with prior art, reduce energy loss.
Description
Technical field
The invention belongs to field of biotechnology, and in particular to a kind of non-thermal sterilization method of ultrasonic in combination HIGH PRESSURE TREATMENT has
Effect induction gemma release DPA is to reduce its heat resistance.
Background technique
DPA, 2, dipicolimic acid 2.
Bacillus cereus is that one kind can give birth to sporiferous bacterium, in the range of 4-55 DEG C of temperature, pH 3.3-4.9 all
It can survive, and can quickly breed under refrigerated conditions, produce vomitoxin and diarrhea toxin.Bacillus cereus is deposited extensively
It is in environment, be easy to cause food pollution, causes to poison by food.In addition, dairy products are highly susceptible to pollute.Infant's exempts from
Epidemic disease power is lower.Once there was 1 food poisoning in April, 2007 in Tonglu, Zhejiang, and 7~August age consumption by infants infant matches
Diarrhea occurs after square milk powder, is the pathogenic bacteria for causing the infantile diarrhea, bacterium through the bacillus cereus in detection discovery milk powder
Falling sum is only 160CFU/mL.
Gemma is that the suspend mode for the round or ellipse that thallus is formed in extreme circumstances constructs, compact structure, and inside contains
Water is low, and metabolic rate is low, is resistant to a variety of extreme environmental conditions, including heat treatment, ultraviolet light, acid, alkali, aldehydes, enzyme,
Phenols, alkylating reagent etc..Gemma has very strong resistance, and such as pasteurize of common sterilizing means can not be killed
It goes out.Simultaneously growth and breeding can be sprouted, under certain condition so as to cause serious food poisoning.It, need to be to wax to guarantee food safety
Sample bacillus spore is controlled.
Studies have shown that dipicolimic acid 2 (DPA) content is high 2 in gemma, the aqueous of gemma core can be greatly reduced
Amount, so that inhereditary material in gemma core is protected to destroy from external world's processing, it is vital to maintain the resistance of gemma to play
Effect.Therefore the release of DPA is to reduce a committed step of gemma resistance.Currently, only super-pressure (200MPa or more)
Processing can open DPA ion channel, accelerate DPA release, to reduce gemma resistance.But pressure needed for processing is higher,
The high requirements on the equipment, it is at high cost.
To sum up, it needs to be further improved the prior art.
Summary of the invention
The technical problem to be solved by the present invention is to propose that a kind of induction gemma carried out using ultrasonic in combination high pressure discharges DPA
The method for reducing heat resistance.
In order to solve the above technical problems, the present invention proposes a kind of method that induction gemma release DPA reduces heat resistance, including
Following steps:
By product subject to sterilization under room temperature (20~30 DEG C), in the moulding pressure of 350~450kPa, carrying out amplitude is
100% (114 μm), frequency are 25~35min of ultrasonic treatment of 18~22khz.
The improvement of the method for heat resistance is reduced as present invention induction gemma release DPA:
The moulding pressure is 400kPa.
The further improvements in methods of heat resistance are reduced as present invention induction gemma release DPA:
The frequency is 20khz.
The further improvements in methods of heat resistance are reduced as present invention induction gemma release DPA:
The ultrasonic treatment 30min.
Compared with prior art, the technical advantages of the present invention are that:
The present invention discharges DPA using method induction gemma associated with ultrasonic wave and high pressure (350~450kPa)；With it is individual
HIGH PRESSURE TREATMENT, ultrasonic treatment are compared, and acoustic pressure Combined Treatment can induce gemma to discharge most of DPA release, reduce gemma
Resistance.
Pressure needed for ultrasonic combined HIGH PRESSURE TREATMENT is not high, and energy loss can be reduced at the same time by reducing equipment cost,
Play reduce cost, consumption reduction, environmental protection important function.
Specific embodiment
The present invention is described further combined with specific embodiments below, but protection scope of the present invention is not limited in
This.
The method that embodiment 1, induction gemma release DPA reduce heat resistance: product subject to sterilization is in the pressurization of 350~450kPa
Pressure, progress amplitude are 100% (114 μm), and frequency is 25~35min of ultrasonic treatment of 20 ± 2khz.
The gemma that the present invention uses is the ATCC14579 bacillus cereus of Qingdao Hai Bo Biotechnology Co., Ltd.This
Embodiment takes the 1 × 10 of 30mL7.3The spore solution of CFU/mL is as product subject to sterilization.
By product subject to sterilization in the moulding pressure of 400kPa in the present embodiment, progress amplitude is 100% (114 μm), frequency
For the ultrasonic treatment of 20khz, the processing time is 30min.
Note: above-mentioned ultrasound occurs simultaneously with pressurized operation, such as the invention application No. is 201810421105.2 can be used
The compound inactivation brood cell's equipment of pressure thermoacoustic provided by patent " the pressure compound inactivation brood cell method and apparatus of thermoacoustic " realizes aforesaid operations.
Comparative example 1-1, the moulding pressure of step 2 in embodiment 1 is changed to 100kPa (normal pressure), that is, only carry out at ultrasound
Reason；Remaining is equal to embodiment 1.
Comparative example 1-2, cancel the ultrasonic treatment of embodiment 1, that is, only carry out pressurized treatments；Remaining is equal to embodiment 1.
Comparative example 1-3, duration ultrasonic in 1 step 2 of embodiment is changed to 10min, remaining is equal to embodiment 1.
Comparative example 1-4, duration ultrasonic in 1 step 2 of embodiment is changed to 40min, remaining is equal to embodiment 1.
Experiment 1, induction gemma release DPA reduce gemma survival condition measurement after heat resistance:
Resulting solution takes 0.1mL to be spread evenly across 20mL nutrient agar (peptone after above-described embodiment 1 is sterilized
10.0g/L, powdered beef 3.0g/L, sodium chloride 5.0g/L, agar 15.0g/L, pH value 7.3 ± 0.1) 0.1% soluble shallow lake is added
Powder, 37 DEG C of constant temperature incubations count after for 24 hours, and blank control (untreated spore solution) is arranged and carries out identical experiment, specifically
Data are as shown in table 1.
Note: spore solution, that is, 30mL 1 × 107.3The spore solution of CFU/mL.
The step of above-mentioned all resulting solution of comparative example are tested in accordance with the above-mentioned embodiment 1 is successively tested, specifically
Data are as shown in table 1.
Plate technique clump count unit after 1 different disposal of table: log (CFU/mL)
| Clump count/log (CFU/mL) | |
| Blank control | 7.30±0.02 |
| Embodiment 1 | 7.00±0.02 |
| Comparative example 1-1 | 7.18±0.04 |
| Comparative example 1-2 | 7.32±0.01 |
| Comparative example 1-3 | 7.16±0.06 |
| Comparative example 1-4 | 7.01±0.05 |
Experiment 2: gemma DPA burst size before and after the processing:
Positive control 1 is arranged: by spore solution through 121 DEG C, 100kPa steam sterilizing handles 30min；
With the leakage rate of DPA in 1 processing gained each group solution of colorimetric method for determining experiment, it is used as and is measured using iron ammonium sulfate
The color developing agent of DPA.
Color developing agent of the iron ammonium sulfate as measurement DPA has preferable measurement effect, is capable of detecting when micro in solution
DPA exist.Simultaneously in the pH value range of 4.0-6.0, using ascorbic acid (Vc), is reacted with DPA, be combined into a kind of stabilization
Compound, have characteristic absorption peak at 440nm.
Gemma DPA burst size unit after 2 different disposal of table: μ g/mL
| Clump count/log (CFU/mL) | |
| Positive control 1 | 311.32±0.60 |
| Blank control | 0.54±0.03 |
| Embodiment 1 | 288.01±0.93 |
| Comparative example 1-1 | 51.59±0.17 |
| Comparative example 1-2 | 1.98±0.23 |
| Comparative example 1-3 | 104.52±0.45 |
| Comparative example 1-4 | 290.22±0.67 |
By the result of table 2 as it can be seen that individually ultrasonic treatment is only capable of the DPA of release 33.99%；The HIGH PRESSURE TREATMENT of independent 400kPa
Gemma cannot be induced to discharge DPA；Ultrasonic in combination HIGH PRESSURE TREATMENT can induce DPA significantly to discharge, and can discharge after handling 30min
92.54% DPA, ultrasonic in combination high pressure have preferable synergy to induction DPA release.Extend processing time loss largely to provide
Source, but DPA burst size does not dramatically increase.
Note: DPA burst size 311.32 ± 0.60 is DPA total amount in positive control 1, to calculate embodiment 1 and each right
Ratio processing after DPA burst size ratio, e.g., comparative example 1-1 individually be ultrasonically treated in DPA burst size be 51.59 ± 0.17,
51.59/311.32*100% it is equal to 33.99%, to learn the DPA for being individually ultrasonically treated and being only capable of release 33.99%.
Experiment 3: the measurement of gemma heat resistance:
Positive control 2 is arranged: by spore solution in 80 DEG C, the moulding pressure of 400kPa, progress amplitude is 100% (114 μ
M), frequency is the ultrasonic treatment of 20khz, and the processing time is 30min.
1 processing gained each group solution water bath with thermostatic control at 80 DEG C of experiment is heat-treated 20min, takes out and is dropped with cold bath
Temperature detects (with experiment 1) gemma amount of survival.By heat treatment front and back Number of spores, it is strong and weak to compare gemma heat resistance.
Gemma surviving colonies number unit after table 3 is heat-treated: log (CFU/mL)
| Clump count/log (CFU/mL) | |
| Positive control 2 | 4.47±0.23 |
| Blank control | 7.21±0.04 |
| Embodiment 1 | 3.76±0.05 |
| Comparative example 1-1 | 6.18±0.00 |
| Comparative example 1-2 | 7.01±0.02 |
| Comparative example 1-3 | 5.84±0.03 |
| Comparative example 1-4 | 3.78±0.03 |
To sum up, ultrasonic in combination high pressure can induce bacillus cereus gemma to discharge DPA, so that it is anti-that its heat is greatly lowered
Property.
The above list is only a few specific embodiments of the present invention for finally, it should also be noted that.Obviously, this hair
Bright to be not limited to above embodiments, acceptable there are many deformations.Those skilled in the art can be from present disclosure
All deformations for directly exporting or associating, are considered as protection scope of the present invention.
Claims (4)
1. the method for inducing gemma release DPA to reduce heat resistance, it is characterized in that the following steps are included:
At room temperature by product subject to sterilization, in the moulding pressure of 350~450kPa, carrying out amplitude is 100%, frequency is 18~
25~35min of ultrasonic treatment of 22khz.
2. the method that induction gemma release DPA reduces heat resistance according to claim 1, it is characterized in that:
The moulding pressure is 400kPa.
3. the method that induction gemma release DPA reduces heat resistance according to claim 2, it is characterized in that:
The frequency is 20khz.
4. the method that any induction gemma release DPA reduces heat resistance according to claim 1~3, it is characterized in that:
The ultrasonic treatment 30min.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3968248A (en) * | 1971-06-21 | 1976-07-06 | Wave Energy Systems, Inc. | Method and sporicidal compositions for synergistic disinfection or sterilization |
| JP2002191336A (en) * | 2000-12-27 | 2002-07-09 | Shimadzu Corp | Method and system for treating liquid material |
| CN106213115A (en) * | 2016-07-27 | 2016-12-14 | 浙江大学 | A kind of method effectively killing the sublethal damage bacterium produced in heat-sterilization process |
| CN108740684A (en) * | 2018-04-27 | 2018-11-06 | 浙江大学 | Press the compound inactivation brood cell's method and apparatus of thermoacoustic |
-
2018
- 2018-12-14 CN CN201811531062.XA patent/CN109757645A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3968248A (en) * | 1971-06-21 | 1976-07-06 | Wave Energy Systems, Inc. | Method and sporicidal compositions for synergistic disinfection or sterilization |
| JP2002191336A (en) * | 2000-12-27 | 2002-07-09 | Shimadzu Corp | Method and system for treating liquid material |
| CN106213115A (en) * | 2016-07-27 | 2016-12-14 | 浙江大学 | A kind of method effectively killing the sublethal damage bacterium produced in heat-sterilization process |
| CN108740684A (en) * | 2018-04-27 | 2018-11-06 | 浙江大学 | Press the compound inactivation brood cell's method and apparatus of thermoacoustic |
Non-Patent Citations (4)
| Title |
|---|
| WILLIAM H. COLEMAN等: "How Moist Heat Kills Spores of Bacillus subtilis", 《JOURNAL OF BACTERIOLOGY》 * |
| 中国科学技术情报研究所重庆分所: "《国外遗传学研究专集》", 31 October 1979, 科学技术文献出版社重庆分社 * |
| 王涛: "DPA 对细胞壁缺陷枯草芽胞杆菌耐热性影响的研究", 《贵州医药》 * |
| 薛广波: "《现代消毒学》", 30 July 2002, 人民军医出版社 * |
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Application publication date: 20190517 |