CN114574543A - High-temperature-resistant lactic acid bacteria breeding method - Google Patents
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Abstract
The invention discloses a high-temperature-resistant lactic acid bacteria breeding method, which comprises the following steps: s1, preparing a culture medium; MRS culture medium: the method comprises the steps of firstly preparing a culture medium, then carrying out primary screening on the lactobacillus, identifying the lactobacillus, carrying out high-temperature resistant breeding, measuring a growth curve, researching acid production capacity and the like, and finally obtaining the high-temperature resistant lactobacillus strain, wherein the acid production capacity of the lactobacillus with different temperature tolerance limits is researched through a comparison experiment in the process, and the breeding box used in the invention for high temperature resistant breeding passes through the rotatable door, guarantee when the culture dish is taken out or is sent back that the breed selection case is isolated with external space, can guarantee can not pollute other culture dishes when taking out the target culture dish, improved the accuracy and the persuasion of experiment.
Description
Technical Field
The invention belongs to the technical field of bioengineering, and particularly relates to a high-temperature-resistant lactic acid bacteria breeding method.
Background
The method has the advantages that the research of excellent strain breeding and fermentation process control and the proposal of a new online separation process have remarkable promotion effect on the realization of industrial large-scale production of lactic acid, have certain reference effect on relevant microbial fermentation production, and combine physicochemical mutagenesis and a chemostat in the breeding research of high-temperature-resistant lactic acid bacteria, thereby greatly improving the screening efficiency of forward mutant plants, reducing the workload in strain screening, and having certain reference effect on other strains with tolerance to screening environment. The proper growth temperature of the lactic acid bacteria is about 37 ℃, the temperature of the growth environment of the lactic acid bacteria needs to be increased when the high-temperature resistant breeding of the lactic acid bacteria is carried out, most of the lactic acid bacteria die in the process, so the growth condition of the lactic acid bacteria needs to be observed in time, when the growth state of the lactic acid bacteria changes, the lactic acid bacteria needs to be taken out from the high-temperature environment in time for further confirmation and research, but the lactic acid bacteria are anaerobic bacteria, and the lactic acid bacteria which are not high-temperature resistant and are taken out frequently can influence other growth environments of the lactic acid bacteria.
Disclosure of Invention
In order to solve the problems in the prior art, the invention aims to provide a high-temperature-resistant lactic acid bacteria breeding method.
A breeding method of high-temperature-resistant lactic acid bacteria is characterized by comprising the following steps:
s1, preparing a culture medium; MRS culture medium: 10.0g of peptone, 10.0g of beef extract, 5.0g of yeast extract, 2.0g of diammonium hydrogen citrate, 20.0g of driving friction wheel of glucose, 1.0mL of Tween 80, 5.0g of sodium acetate, 2.0g of dipotassium hydrogen phosphate, 0.58g of magnesium sulfate, 0g of manganese sulfate, 25g of transmission gear, 18.0g of sliding rail of agar and 1000mL of distilled water, sterilizing a bracket for 15min at 21 ℃ of a first rotating shaft of a rotating door 1 by using an autoclave, adjusting the pH to 6.2-6.6, and culturing a seed culture medium: liquid MRS culture medium;
s2, primary screening of lactic acid bacteria; transferring 1mL of aquaculture water serving as a bacteria source into a test tube containing 9mL of sterilized normal saline, shaking uniformly, diluting a sample, sucking 0.1mL of diluent, adding the diluent to an MRS culture medium containing 0.5% of calcium carbonate, performing anaerobic culture on a power module at 37 ℃ for 24-48 hours by using a synchronous belt, observing the shape after the bacteria form, selecting bacterial colonies with calcium-soluble rings around, and repeatedly scribing on an MRS flat plate until single bacterial colonies are obtained;
s3, identifying lactic acid bacteria; the morphological identification of the bacterial colony is carried out by bacterial colony observation, gram staining, spore staining and flagellum staining, and the physiological and biochemical identification of the lactobacillus is carried out by indole experiment, catalase experiment, nitrate reduction experiment and gelatin liquefaction experiment;
s4, high-temperature resistant breeding; inoculating lactobacillus to a seed culture medium, placing the seed culture medium into a sterile and anaerobic breeding box, adjusting the initial temperature of the breeding box to be 37 ℃ of a power module, adjusting the temperature once every 6 hours, sequentially adjusting the temperature to be 40 ℃, 45 ℃, 50 ℃, 55 ℃ and 60 ℃ of a servo motor, observing the colony morphology every 2 hours during the period, and moving the culture medium which is obviously lagged behind the growth state of other colonies and is picked out to a constant-temperature culture box with the same temperature as that of the current breeding box;
s5, measuring a growth curve; selecting lactobacillus strains in constant-temperature incubators at different temperatures, inoculating the lactobacillus strains to a 30mL liquid MRS culture medium of a driven gear, measuring the OD value of the lactobacillus strains at intervals of 2h, and taking time as a horizontal coordinate and the OD value as a vertical coordinate to obtain a growth curve suitable for the lactobacillus strains at different temperatures;
s6, acid-producing capacity research; selecting lactic acid bacteria at corresponding temperature according to the growth curve of the lactic acid bacteria, culturing the optimum growth time according to the growth curve, inoculating the lactic acid bacteria into fresh milk, measuring the pH value of the milk every 2 hours, and obtaining the acid production capacity curve of the lactic acid bacteria by taking the time as the abscissa and the pH value as the ordinate;
s7, obtaining high-temperature-resistant lactic acid bacteria; according to the acid production capability of the lactic acid bacteria suitable for different temperatures, the highest temperature suitable for the lactic acid bacteria is compared to obtain the lactic acid bacteria with high temperature resistance and acid production capability in a normal range, and the lactic acid bacteria are selected as the later-stage research objects.
Preferably, in step S1, the lactic acid bacteria metabolize lactic acid and then the pH is lowered to change the color from green to yellow-green.
As a preferable mode of the present invention, in step S2, the samples are diluted to 10-5, 10-6 and 10-7 before being dropped on the MRS medium, respectively.
Preferably, in steps S4 and S5, the incubator includes a plurality of chambers having different temperatures, and the temperature of the petri dish taken out of the breeding chamber is maintained.
Preferably, in step S4, the seed selection box includes a box body, a plurality of support plates are disposed in the box body, a plurality of trays are disposed between two adjacent support plates, each tray is slidably connected to the box body or the support plate through two sliding grooves, and a culture dish is placed on each tray.
Preferably, the breeding box further comprises a plurality of fixing plates, each fixing plate is fixedly connected with the box body or the supporting plate, each fixing plate is rotatably provided with a second rotating shaft, each second rotating shaft is fixedly provided with a rotating door and a rotating gear, one side of each rotating gear is meshed with a sector gear, each sector gear is fixedly arranged on a third rotating shaft, each third rotating shaft is rotatably connected with the box body or the supporting plate, and each third rotating shaft is further fixedly provided with a transmission friction wheel and a driven gear.
Preferably, a driving gear is engaged with one side of each driven gear, each driving gear is fixedly arranged on a fourth rotating shaft, each fourth rotating shaft is rotatably connected with the box body or the supporting plate, a driving pulley is fixedly arranged on each fourth rotating shaft, each driving pulley is driven by a synchronous belt to form a driving pulley, each driving pulley is fixedly arranged on a fifth rotating shaft, each fifth rotating shaft is rotatably connected with the box body or the supporting plate, and a transmission gear is fixedly arranged on each fifth rotating shaft.
Preferably, one side of each tray is provided with a row of teeth, and each transmission gear is meshed with the teeth on the tray.
Preferably, the breeding box further comprises a plurality of slide rails, two adjacent slide rails are connected through a guide groove, a power module is arranged in the uppermost slide rail in a sliding manner, a servo motor is fixedly connected to one side of the power module, a sixth rotating shaft is controlled by one end of the servo motor, a rotating wheel is fixedly arranged on the sixth rotating shaft, a driving motor is fixedly connected to one side, away from the servo motor, of the power module, a first rotating shaft is controlled by one end of the driving motor, a driving friction wheel is fixedly arranged on the first rotating shaft, and the driving friction wheel is in friction transmission with one of the driving friction wheels.
Preferably, the guide grooves are distributed on two sides of the slide rail in a staggered manner, and the uppermost guide groove is located on one side of the slide rail close to the power supply module.
The beneficial effects of the invention are as follows: the invention relates to a high-temperature-resistant lactic acid bacteria breeding method, which comprises the steps of preparing a culture medium, primarily screening lactic acid bacteria, identifying lactic acid bacteria, breeding at high temperature resistance, measuring a growth curve, researching acid production capacity and the like, and finally obtaining the high-temperature-resistant lactic acid bacteria strain.
Drawings
The invention is described in further detail below with reference to the accompanying drawings and specific embodiments.
FIG. 1 is a diagram of the operational steps of the present invention;
FIG. 2 is a schematic structural view of the present invention;
FIG. 3 is a schematic view of the invention taken in the direction A-A of FIG. 2;
FIG. 4 is a schematic view of the present invention taken in the direction B-B of FIG. 3;
FIG. 5 is an enlarged schematic view at C of FIG. 3 of the present invention;
fig. 6 is an enlarged schematic view at D of fig. 4 according to the present invention.
Detailed Description
The invention is described in detail below with reference to the figures and examples.
In an embodiment of the present invention, referring to fig. 1 to 6, a method for breeding a high temperature resistant lactic acid bacterium is provided, which comprises the following steps:
s1, preparing a culture medium; MRS culture medium: 10.0g of peptone, 10.0g of beef extract, 5.0g of yeast extract, 2.0g of diammonium hydrogen citrate, 20.0g of driving friction wheel of glucose, 1.0mL of Tween 80, 5.0g of sodium acetate, 2.0g of dipotassium hydrogen phosphate, 0.58g of magnesium sulfate, 0g of manganese sulfate, 25g of transmission gear, 18.0g of sliding rail of agar and 1000mL of distilled water, sterilizing a bracket for 15min at 21 ℃ of a first rotating shaft of a rotating door 1 by using an autoclave, adjusting the pH to 6.2-6.6, and culturing a seed culture medium: liquid MRS culture medium;
s2, primary screening of lactic acid bacteria; transferring 1mL of aquaculture water serving as a bacteria source into a test tube containing 9mL of sterilized normal saline, shaking uniformly, diluting a sample, sucking 0.1mL of diluent, adding the diluent to an MRS culture medium containing 0.5% of calcium carbonate, performing anaerobic culture on a power module at 37 ℃ for 24-48 hours by using a synchronous belt, observing the shape after bacteria formation, selecting bacterial colonies with calcium-dissolving rings around, and repeatedly scribing on an MRS plate until single bacterial colonies are obtained;
s3, identifying lactic acid bacteria; the morphological identification of the bacterial colony is carried out by bacterial colony observation, gram staining, spore staining and flagellum staining, and the physiological and biochemical identification of the lactobacillus is carried out by indole experiment, catalase experiment, nitrate reduction experiment and gelatin liquefaction experiment;
s4, high-temperature resistant breeding; inoculating lactobacillus to a seed culture medium, placing the seed culture medium into a sterile and anaerobic breeding box, adjusting the initial temperature of the breeding box to be 37 ℃ of a power supply module, adjusting the temperature once every 6 hours, sequentially adjusting the temperature to be 40 ℃, 45 ℃, 50 ℃, 55 ℃ and 60 ℃ of a servo motor, observing the colony morphology every 2 hours, and moving a culture dish which is obviously lagged behind the growth state of other colonies after being picked out into a constant-temperature culture box;
s5, measuring a growth curve; selecting lactobacillus strains in constant-temperature incubators at different temperatures, inoculating the lactobacillus strains to a 30mL liquid MRS culture medium of a driven gear, measuring the OD value of the lactobacillus strains at intervals of 2h, and taking time as a horizontal coordinate and the OD value as a vertical coordinate to obtain a growth curve suitable for the lactobacillus strains at different temperatures;
s6, acid-producing capacity research; selecting lactic acid bacteria at corresponding temperature according to the growth curve of the lactic acid bacteria, culturing the optimum growth time according to the growth curve, inoculating the lactic acid bacteria into fresh milk, measuring the pH value of the milk every 2 hours, and obtaining the acid production capacity curve of the lactic acid bacteria by taking the time as the abscissa and the pH value as the ordinate;
s7, obtaining high-temperature-resistant lactic acid bacteria; according to the acid production capability of the lactic acid bacteria suitable for different temperatures, comparing the highest temperature suitable for the lactic acid bacteria to obtain the lactic acid bacteria with high temperature resistance and acid production capability in a normal range, and selecting the lactic acid bacteria as a later-stage research object;
it should be noted that, after lactic acid bacteria grow and metabolize lactic acid, the pH value will decrease and the color will change from green to yellow-green, and because the pH value decreases and the effect of antibiotic and anaerobic culture is added, general microorganisms will not grow on the culture medium easily, so it is very easy to identify lactic acid bacteria, pure lactic acid bacteria can be obtained by morphological identification and physiological and biochemical identification, the acid production capability of lactic acid bacteria which can tolerate different temperatures is studied by determining the growth curve of lactic acid bacteria, and the inaccurate acid production capability determination caused by the difference of growth time is avoided.
Advantageously, in step S1, the lactic acid bacteria can be roughly screened by observing their color, which is caused by a decrease in pH after they have metabolized lactic acid, resulting in a change in color from green to yellow-green.
Advantageously, in step S2, the sample is diluted to 10-5、10-6、10-7And then, respectively dripping the lactobacillus into the MRS culture medium, wherein the number of the lactobacillus in the strain is uncertain, and the concentration of the diluted lactobacillus cannot be determined to ensure that the bacterial colony generated by the lactobacillus meets the experimental requirements, so that the diluents with different concentrations are selected for comparison.
Advantageously, in steps S4 and S5, the incubator comprises a plurality of chambers with different temperatures, the temperature of the culture dish taken out from the breeding chamber is kept constant, the temperature of the incubator is the maximum temperature that the lactic acid bacteria can endure, and ensuring that the temperature is constant is an important factor for ensuring the accuracy of the experiment.
Advantageously, in step S4, the breeding box includes a box 11, a plurality of supporting plates 17 are disposed in the box 11, a plurality of trays 14 are disposed between two adjacent supporting plates 17, each tray 14 is slidably connected to the box 11 or the supporting plates 17 through two sliding grooves 13, a culture dish 16 is disposed on each tray 14, and the taking and placing of the culture dish is controlled by using a mechanical transmission manner, so that the pollution to the lactic acid bacteria can be effectively avoided without directly contacting the culture dish.
Advantageously, the breeding box further comprises a plurality of fixing plates 41, each fixing plate 41 is fixedly connected with the box body 11 or the supporting plate 17, each fixing plate 41 is rotatably provided with a second rotating shaft 31, each second rotating shaft 31 is fixedly provided with a rotating door 12 and a rotating gear 32, one side of each rotating gear 32 is engaged with a sector gear 33, each sector gear 33 is fixedly provided on a third rotating shaft 28, each third rotating shaft 28 is rotatably connected with the box body 11 or the supporting plate 17, each third rotating shaft 28 is further fixedly provided with a transmission friction wheel 29 and a driven gear 30, the rotating door 12 is controlled by the sector gear 33, and when the sector gear 33 and the rotating gear 32 are not engaged, the rotating door 12 can be quickly closed under the action of gravity.
Beneficially, a driving gear 35 is engaged with one side of each driven gear 30, each driving gear 35 is fixedly arranged on a fourth rotating shaft 36, each fourth rotating shaft 36 is rotatably connected with the case 11 or the supporting plate 17, each fourth rotating shaft 36 is further fixedly provided with a driving pulley 34, each driving pulley 34 is driven by a synchronous belt 24 to form a driving pulley 26, each driving pulley 26 is fixedly arranged on a fifth rotating shaft 27, each fifth rotating shaft 27 is rotatably connected with the case 11 or the supporting plate 17, each fifth rotating shaft 27 is further fixedly provided with a transmission gear 25, the transmission of the synchronous belt 26 ensures that the rotating door 12 can be opened and closed at the first time when the culture dish 16 is taken and placed, and ensures that other culture dishes 16 are not polluted.
Advantageously, one side of each tray 14 is provided with a row of teeth, and each transmission gear 25 is engaged with the teeth on the tray 14 to smoothly transfer the culture dish 16 through the teeth on the tray 14, so that the living space of the strains in the culture dish is not severely changed.
Advantageously, the breeding box further comprises a plurality of slide rails 18, two adjacent slide rails 18 are connected through a guide groove 23, a power supply module 37 is arranged in the uppermost one of the slide rails 18 in a sliding manner, one side of the power module 37 is fixedly connected with a servo motor 40, one end of the servo motor 40 is controlled by a sixth rotating shaft 39, a rotating wheel 38 is fixedly arranged on the sixth rotating shaft 39, a driving motor 22 is fixedly connected to one side of the power module 37 far away from the servo motor 40, one end of the driving motor 22 is controlled by a first rotating shaft 21, a driving friction wheel 20 is fixedly arranged on the first rotating shaft 21, the driving friction wheel 20 is in friction transmission with one of the transmission friction wheels 29, a single power source is adopted, the cost is greatly reduced, and the transmission friction wheels 29 are used for transmission, so that the conversion transmission parts are prevented from being severely abraded.
Advantageously, the guiding grooves 23 are distributed on two sides of the sliding rail 18 in a staggered manner, the uppermost one of the guiding grooves 23 is located on one side of the sliding rail 18 close to the power module 37, the guiding grooves 23 are not distributed on the same side of the sliding rail 18, so that the servo motor 40 can be prevented from directly sliding from the uppermost sliding rail 18 to the lowermost sliding rail 18, and the servo motor 40 can only slide along the sliding rail layer by layer.
The working principle of the breeding box is as follows:
the servo motor 40 and the driving motor 22 are controlled to work by controlling the power module 37, the servo motor 40 is started, the servo motor 40 controls the sixth rotating shaft 39 to rotate, the sixth rotating shaft 39 drives the rotating wheel 38 to rotate, the rotating wheel 38 rolls along the sliding rail 18 and the guide groove 23 until the driving motor 22 slides to the culture dish 16 to be taken away, the servo motor 40 is closed, and the driving motor 22 is started;
the driving motor 22 controls the first rotating shaft 21 to rotate, the first rotating shaft 21 drives the driving friction wheel 20 to rotate, the driving friction wheel 20 drives the driving friction wheel 29 through friction transmission, the driving friction wheel 29 drives the third rotating shaft 28 to rotate, the third rotating shaft 28 drives the sector gear 33 and the driven gear 30 to rotate, the rotating gear 32 is driven to rotate through meshing, the rotating gear 32 drives the second rotating shaft 31 to rotate, the second rotating shaft 31 drives the rotating door 12 to rotate, meanwhile, the driven gear 30 drives the driving gear 35 to rotate through meshing, the driving gear 35 drives the fourth rotating shaft 36 to rotate, the fourth rotating shaft 36 drives the driving belt wheel 34 to rotate, the driving belt wheel 34 drives the driving belt wheel 26 to rotate through the synchronous belt 24, the driving belt wheel 26 drives the fifth rotating shaft 27 to rotate, the fifth rotating shaft 27 drives the driving gear 25 to rotate, and the driving gear 25 drives the tray 14 to slide along the chute 13 through meshing, when the culture dish 16 slides to the other side of the rotating door 12, the driving motor 22 is closed;
at this time, the sector gear 33 is no longer meshed with the rotating gear 32, the rotating gear 32 is not restrained under the action of the gravity of the rotating door 12, the rotating door 12 drives the second rotating shaft 31 to rotate until the rotating door 12 returns to the vertical state, and the box body 11 is manually opened to take the target culture dish 16;
the driving motor 22 is started reversely, the driving motor 22 controls the first rotating shaft 21 to rotate, the first rotating shaft 21 drives the driving friction wheel 20 to rotate, the driving friction wheel 20 drives the driving friction wheel 29 through friction, the driving friction wheel 29 drives the third rotating shaft 28 to rotate, the third rotating shaft 28 drives the sector gear 33 and the driven gear 30 to rotate, the rotating gear 32 is driven to rotate through meshing, the rotating gear 32 drives the second rotating shaft 31 to rotate, the second rotating shaft 31 drives the rotating door 12 to rotate, meanwhile, the driven gear 30 drives the driving gear 35 to rotate through meshing, the driving gear 35 drives the fourth rotating shaft 36 to rotate, the fourth rotating shaft 36 drives the driving pulley 34 to rotate, the driving pulley 34 drives the driving pulley 26 to rotate through the synchronous belt 24, the driving pulley 26 drives the fifth rotating shaft 27 to rotate, the fifth rotating shaft 27 drives the driving gear 25 to rotate, the driving gear 25 drives the tray 14 to slide along the chute 13 to an initial position through meshing, the driving motor 22 is turned off, at this time, the sector gear 33 is no longer meshed with the rotating gear 32, the rotating gear 32 is not restrained under the action of the gravity of the rotating door 12, and the rotating door 12 drives the second rotating shaft 31 to rotate until the rotating door 12 returns to the vertical state;
the invention is not limited to the above alternative embodiments, and any other various forms of products can be obtained by anyone in the light of the present invention, but any changes in shape or structure thereof, which fall within the scope of the present invention as defined in the claims, fall within the scope of the present invention.
Claims (10)
1. A breeding method of high-temperature-resistant lactic acid bacteria is characterized by comprising the following steps:
s1, preparing a culture medium; MRS culture medium: 10.0g of peptone, 10.0g of beef extract, 5.0g of yeast extract, 2.0g of diammonium hydrogen citrate, 20.0g of driving friction wheel of glucose, 1.0mL of Tween 80, 5.0g of sodium acetate, 2.0g of dipotassium hydrogen phosphate, 0.58g of magnesium sulfate, 0g of manganese sulfate, 25g of transmission gear, 18.0g of sliding rail of agar and 1000mL of distilled water, sterilizing a bracket for 15min at 21 ℃ of a first rotating shaft of a rotating door 1 by using an autoclave, adjusting the pH to 6.2-6.6, and culturing a seed culture medium: liquid MRS culture medium;
s2, primary screening of lactic acid bacteria; transferring 1mL of aquaculture water serving as a bacteria source into a test tube containing 9mL of sterilized normal saline, shaking uniformly, diluting a sample, sucking 0.1mL of diluent, adding the diluent to an MRS culture medium containing 0.5% of calcium carbonate, performing anaerobic culture on a power module at 37 ℃ for 24-48 hours by using a synchronous belt, observing the shape after bacteria formation, selecting bacterial colonies with calcium-dissolving rings around, and repeatedly scribing on an MRS plate until single bacterial colonies are obtained;
s3, identifying lactic acid bacteria; the morphological identification of the bacterial colony is carried out by bacterial colony observation, gram staining, spore staining and flagellum staining, and the physiological and biochemical identification of the lactobacillus is carried out by indole experiment, catalase experiment, nitrate reduction experiment and gelatin liquefaction experiment;
s4, high-temperature resistant breeding; inoculating lactobacillus to a seed culture medium, placing the seed culture medium into a sterile and anaerobic breeding box, adjusting the initial temperature of the breeding box to be 37 ℃ of a power module, adjusting the temperature once every 6 hours, sequentially adjusting the temperature to be 40 ℃, 45 ℃, 50 ℃, 55 ℃ and 60 ℃ of a servo motor, observing the colony morphology every 2 hours during the period, and moving the culture medium which is obviously lagged behind the growth state of other colonies and is picked out to a constant-temperature culture box with the same temperature as that of the current breeding box;
s5, measuring a growth curve; selecting lactobacillus strains in constant-temperature incubators at different temperatures, inoculating the lactobacillus strains to a 30mL liquid MRS culture medium of a driven gear, measuring the OD value of the lactobacillus strains at intervals of 2h, and taking time as a horizontal coordinate and the OD value as a vertical coordinate to obtain a growth curve suitable for the lactobacillus strains at different temperatures;
s6, acid-producing capacity research; selecting lactic acid bacteria at corresponding temperature according to the growth curve of the lactic acid bacteria, culturing the optimum growth time according to the growth curve, inoculating the lactic acid bacteria into fresh milk, measuring the pH value of the milk every 2 hours, and obtaining the acid production capacity curve of the lactic acid bacteria by taking the time as the abscissa and the pH value as the ordinate;
s7, obtaining high-temperature-resistant lactic acid bacteria; according to the acid production capability of the lactic acid bacteria suitable for different temperatures, the highest temperature suitable for the lactic acid bacteria is compared to obtain the lactic acid bacteria with high temperature resistance and acid production capability in a normal range, and the lactic acid bacteria are selected as the later-stage research objects.
2. The method for breeding high-temperature-resistant lactic acid bacteria according to claim 1, wherein: in step S1, the lactic acid bacteria grow to metabolize the lactic acid and then the pH is lowered resulting in a color change from green to yellow-green.
3. The method for breeding high-temperature-resistant lactic acid bacteria according to claim 1, wherein: in step S2, the sample is diluted to 10-5、10-6、10-7Then, the mixture is respectively dripped on an MRS culture medium.
4. The method for breeding high-temperature-resistant lactic acid bacteria according to claim 1, wherein: in steps S4 and S5, the incubator includes a plurality of chambers having different temperatures, and the temperature of the culture dish taken out from the breeding chamber is kept constant.
5. The method for breeding high-temperature-resistant lactic acid bacteria according to claim 1, wherein: in step S4, the seed selection box includes a box body, a plurality of support plates are disposed in the box body, a plurality of trays are disposed between two adjacent support plates, each tray is slidably connected to the box body or the support plate through two sliding grooves, and a culture dish is placed on each tray.
6. The breeding method of high temperature resistant lactic acid bacteria according to claim 5, characterized in that: the breeding case still includes a plurality of fixed plates, every the fixed plate all with the box perhaps backup pad fixed connection, every it is equipped with the second axis of rotation, every to rotate on the fixed plate fixed rotating door and the running gear of being equipped with in the second axis of rotation, every the meshing of running gear one side has sector gear, every sector gear is fixed to be set up in the third axis of rotation, every the third axis of rotation with the box perhaps the backup pad rotates to be connected, every still fixed transmission friction pulley and the driven gear of being equipped with in the third axis of rotation.
7. The breeding method of high temperature resistant lactic acid bacteria according to claim 6, characterized in that: every driven gear one side meshing has drive gear, every drive gear is fixed to be set up in the fourth axis of rotation, every the fourth axis of rotation with the box perhaps the backup pad rotates to be connected, every still fixedly in the fourth axis of rotation is equipped with driving pulley, every driving pulley has driving pulley, every through synchronous belt drive driving pulley is fixed to be set up in the fifth axis of rotation, every the fifth axis of rotation with the box perhaps the backup pad rotates to be connected, every still fixedly in the fifth axis of rotation is equipped with drive gear.
8. The breeding method of high temperature resistant lactic acid bacteria according to claim 7, characterized in that: one side of each tray is provided with a row of teeth, and each transmission gear is meshed with the teeth on the tray.
9. The breeding method of high temperature resistant lactic acid bacteria according to claim 5, characterized in that: the breeding case still includes a plurality of slide rails, adjacent two connect through the guiding groove between the slide rail, one of the top slide in the slide rail and be equipped with power module, one side fixedly connected with servo motor of power module, servo motor one end control system has the sixth axis of rotation, the fixed rotation wheel that is equipped with in the sixth axis of rotation, power module keeps away from servo motor's one side fixedly connected with driving motor, driving motor one end control system has first axis of rotation, the fixed drive friction pulley that is equipped with in the first axis of rotation, drive friction pulley and one of them transmission friction pulley friction drive.
10. The breeding method of high temperature resistant lactic acid bacteria according to claim 9, characterized in that: the guiding grooves are distributed on two sides of the sliding rail in a staggered mode, and the guiding groove on the uppermost side is located on one side, close to the power module, of the sliding rail.
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