CN112023066A - Sterilization kettle and culture method of aspergillus niger applying same - Google Patents
Sterilization kettle and culture method of aspergillus niger applying same Download PDFInfo
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- CN112023066A CN112023066A CN202010921716.0A CN202010921716A CN112023066A CN 112023066 A CN112023066 A CN 112023066A CN 202010921716 A CN202010921716 A CN 202010921716A CN 112023066 A CN112023066 A CN 112023066A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
- A61L2/02—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using physical phenomena
- A61L2/04—Heat
- A61L2/06—Hot gas
- A61L2/07—Steam
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
- A61L2/26—Accessories or devices or components used for biocidal treatment
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/02—Preparation of other alcoholic beverages by fermentation
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2202/00—Aspects relating to methods or apparatus for disinfecting or sterilising materials or objects
- A61L2202/10—Apparatus features
- A61L2202/11—Apparatus for generating biocidal substances, e.g. vaporisers, UV lamps
Abstract
The application discloses a sterilization kettle and a culture method of Aspergillus niger by using the sterilization kettle, the sterilization kettle comprises a frame and a kettle body, the kettle body is provided with a material port for loading and unloading, the kettle body is provided with a material baffle plate for closing the material port, the kettle body is rotationally connected with the frame, so that the material port rotates towards the direction close to or far away from the ground, the kettle body is communicated with a steam inlet pipe for supplying steam into the kettle body, the steam inlet pipe is provided with a first valve for adjusting the water vapor flux in the steam inlet pipe, the kettle body is communicated with a steam exhaust pipe for exhausting the gas in the kettle body, the steam exhaust pipe is provided with a second valve for adjusting the water vapor flux in the steam exhaust pipe, and the rack is provided with a driving assembly for driving the kettle body to rotate, so that the aim of reducing the survival rate of the mixed bacteria in the material is fulfilled; the application also provides a culture method of the aspergillus niger, which has the effects of low infection rate and simple operation.
Description
Technical Field
The application relates to the field of aspergillus niger culture, in particular to a sterilization kettle and a culture method of aspergillus niger by using the sterilization kettle.
Background
Aspergillus niger, Deuteromycotina, class Hyphomycetes, order Hyphomycetales, family Moniliaceae, a common species of fungi of the genus Aspergillus. Widely distributed throughout the world in foodstuffs, vegetable products and soils. Is an important fermentation industrial strain, and can produce amylase, acid protease, cellulase, pectinase, glucose oxidase, citric acid, gluconic acid, gallic acid and the like.
At present, when aspergillus niger is used as a zymocyte of white spirit, culture and inoculation are needed, a common culture medium in a factory is a mixture of bran and water, and the mixture of the bran and the water needs to be sterilized before the culture medium is used, so that the influence of impurities such as other strains in the mixture on the aspergillus niger is reduced.
In the traditional process, a worker generally places a mixture into a pot and continuously introduces steam into the pot to sterilize the mixture, but when materials are more, the materials are stacked in the sterilization mode, so that the situation of incomplete sterilization is easily caused.
Disclosure of Invention
In order to reduce the survival rate of the mixed bacteria in the material, the application provides a sterilization kettle and a culture method of aspergillus niger applying the sterilization kettle.
In a first aspect, the present application provides a sterilization kettle, in order to adopt the following technical scheme:
the utility model provides a sterilization kettle, includes the frame and the cauldron body, offer the material mouth that is used for feeding and unloads on the cauldron body, be provided with on the cauldron body and be used for the closed striker plate of material mouth, the cauldron body rotates with the frame to make the material mouth rotate to the direction that is close to or keeps away from ground, the intercommunication has the admission pipe that is used for supplying into steam to the cauldron body on the cauldron body, be provided with the first valve that is used for adjusting steam flux in the admission pipe on the admission pipe, the intercommunication has the exhaust pipe that is used for gas outgoing in the cauldron body on the cauldron body, be provided with the second valve that is used for adjusting steam flux in the exhaust pipe on the exhaust pipe, be provided with in the frame and be used for driving cauldron body pivoted drive assembly.
By adopting the technical scheme, materials are put into the kettle body from the material port, then the material port is closed by the material baffle plate, the first valve is opened to enable water vapor to be introduced into the kettle body from the vapor inlet pipe, the driving assembly is controlled to enable the driving assembly to drive the kettle body to rotate, the kettle body drives the materials in the kettle body to rotate, the materials are heated and sterilized, and the water vapor is introduced into the kettle body for a long time, so that the pressure in the kettle body is gradually increased, the sterilization effect is further achieved, and the purpose of reducing the survival rate of the mixed bacteria in the materials is further achieved; in addition, the kettle body is connected with the rack in a rotating manner, so that workers can rotate the attached drawings in the direction close to the ground, the material opening faces the ground, and the materials in the kettle body can be poured out by the workers conveniently.
Optionally, fixedly connected with pivot on the cauldron body, and pivot and frame rotate to be connected, the cavity has been seted up on the cauldron body lateral wall, and set up the ventilative hole with the cavity intercommunication on the cauldron body, the chamber that holds has all been seted up at the pivot both ends, hold the chamber lateral wall and set up a plurality of connecting holes that communicate with the cavity with last, pivot one end is stretched out the cauldron body and is connected with steam inlet pipe through rotary joint to make the chamber that holds and steam inlet pipe intercommunication of pivot, the pivot other end stretches out the cauldron body and is connected with steam exhaust pipe through rotary joint, so that the chamber that holds and steam exhaust pipe intercommunication of pivot.
By adopting the technical scheme, the rotary shaft is arranged to realize the rotary connection of the kettle body and the rack, the inner wall of the kettle body is provided with a plurality of steam-permeable holes communicated with the cavity, the contact area of the steam entering the kettle body and the material is increased, and the survival rate of the mixed bacteria in the material is further reduced; the rotating shaft is communicated with the steam inlet pipe and the steam exhaust pipe through the rotating joint, so that the steam inlet pipe and the steam exhaust pipe are not easy to be driven to rotate by the rotation of the rotating shaft.
Optionally, the driving assembly comprises a driving motor, a transmission gear and a driving gear, the driving motor is fixedly connected with the rack, the transmission gear is fixedly connected with the rotating shaft in a coaxial mode, the driving gear is fixedly connected with an output shaft of the driving motor in a coaxial mode, and the driving gear is meshed with the transmission gear.
Through adopting above-mentioned technical scheme, start driving motor for driving motor drives drive gear and rotates, because drive gear and drive gear meshing, consequently drive gear rotates and drives drive gear and rotate, thereby makes the pivot rotate and drives the cauldron body and rotate, thereby realizes the stirring of the internal material of cauldron.
Optionally, the rotating shaft is connected with a shovel plate, and the shovel plate is arranged along the axis of the rotating shaft.
Through adopting above-mentioned technical scheme, the setting of shovel board for when the cauldron body rotated, the shovel board stirred the internal material of cauldron, thereby further improved the homogeneity of disinfecting of material.
Optionally, the shovel plate is connected with the rotating shaft in a rotating mode, the rotating direction of the shovel plate is opposite to that of the rotating shaft, and an adjusting assembly used for adjusting rotation of the shovel plate is arranged on the rotating shaft.
Through adopting above-mentioned technical scheme, the cauldron body rotates, control adjusting part for shovel board and pivot antiport, thereby improved the stirring degree of shovel board to the internal material of cauldron.
Optionally, the adjusting part is including adjusting ring gear, rotating gear and adjusting gear, rotating gear and the coaxial fixed connection of pivot, adjusting gear and rotating gear meshing to rotate with the cavity lateral wall and be connected, adjust the ring gear cover and establish in the adjusting gear outside, and with the adjusting gear meshing, adjust ring gear and shovel board fixed connection, and the axis of just adjusting the ring gear sets up with the rotation axis collineation of shovel board.
Through adopting above-mentioned technical scheme, the pivot rotates to drive the rotating gear and rotate, because adjusting gear and rotating gear meshing, consequently adjusting gear rotates, adjusts the ring gear and rotates and drive the shovel board and rotate along the direction opposite with pivot rotation direction.
In a second aspect, the present application provides a method for culturing aspergillus niger, which adopts the following technical scheme:
a method for culturing Aspergillus niger comprises the following steps: s1, preparing a preliminary culture medium, a first culture medium, a second culture medium and a third culture medium, respectively loading the preliminary culture medium, the first culture medium, the second culture medium and the third culture medium into a preliminary culture dish, a first culture dish, a second culture dish and a third culture dish, and sterilizing the first culture medium, the second culture medium and the third culture medium through a sterilization kettle;
s2, primary culture and separation of Aspergillus niger: after culturing the aspergillus niger in a primary culture medium, separating qualified aspergillus niger in a sterile environment;
s3, inoculating qualified Aspergillus niger separated in S2 into the first culture medium, and shaking the first culture dish to uniformly mix the Aspergillus niger and the first culture medium;
s4, placing a first culture dish containing V48 Aspergillus niger and a first culture medium in an environment of 32-33 ℃, after heat preservation culture, turning over a triangular flask for multiple times, placing the first culture dish in the environment of 32-33 ℃ for continuous heat preservation culture, and finally separating qualified Aspergillus niger in a sterile environment to obtain a first yeast strain;
s5, inoculating the first koji species into a second culture medium, uniformly mixing the first koji species with the second culture medium, stacking the first koji species and the second culture medium in a second culture dish, and then putting the second culture dish containing the first koji species and the second culture medium into an environment at 28-33 ℃ for culture;
s6, spreading the mixture of the first koji in the second culture dish and the second culture medium to the thickness of the mixture of 2.5-3.5cm, and continuously culturing at 28-33 ℃ to obtain the prefabricated koji;
s7, drying the prefabricated seed koji, and separating qualified Aspergillus niger in an aseptic environment to obtain the seed koji;
s8, inoculating the seed starter to a third culture medium for culture, uniformly mixing the seed starter and the third culture medium in a third culture dish, stacking the seed starter and the third culture medium together, and then placing the third culture dish in an environment with the temperature of 28-33 ℃ for culture to obtain a stacking material;
s9, spreading the piled material prepared in the S8 in a third culture dish, wherein the spread thickness is 25-30cm, and placing the third culture dish loaded with the piled material in an environment with the temperature of 28-33 ℃ for culture to obtain the bran koji.
By adopting the technical scheme, the sterilization kettle is adopted to sterilize the triangular flask culture medium, the box culture medium and the pond culture medium, so that the sterilization efficiency of the triangular flask culture medium, the box culture medium and the pond culture medium is improved, and the effect of reducing the survival amount of mixed bacteria in the triangular flask culture medium, the box culture medium and the pond culture medium is achieved; in addition, aspergillus niger is separated and screened for multiple times, so that aspergillus niger strains have the advantages of high purity, strong activity, low infection rate and high saccharification efficiency, the culture method of aspergillus niger is simple to operate, the equipment running rate is low, and resources are saved.
Optionally, in S1, the preliminary culture medium is prepared by mixing the following components in parts by weight: 1100 parts of distilled water 990-type-; uniformly mixing bran and water in a mass ratio of 1:0.5 to prepare a culture material, putting the culture material into a sterilization kettle for sterilization, putting the sterilized culture material into a first culture dish, and finally shaking the first culture dish to cool the culture material in the first culture dish to 30-35 ℃ to prepare the first culture medium; the second culture medium is prepared by uniformly mixing bran and water in a mass ratio of 1:0.5 to prepare a culture material, sterilizing the culture material by using a sterilization kettle, filling the sterilized culture material into a second culture dish, and cooling to 30-35 ℃ of the culture material; the third culture medium is prepared by uniformly mixing bran and rice hull with the mass ratio of 1:0.5:0.1-0.15 with water to prepare a first culture material, sterilizing the first culture material by using a sterilization kettle, loading the sterilized first culture material into a third culture dish, and cooling the first culture material to the temperature of 30-35 ℃.
By adopting the technical scheme, the sterilization kettle is adopted to sterilize each culture medium, so that the survival rate of the mixed bacteria in the material is reduced.
Optionally, in S6, in the process of culturing the second culture medium and the first koji mold, the material in the second culture dish needs to be turned over and/or the second culture dish needs to be turned over.
Through adopting above-mentioned technical scheme, carry out second culture dish upset processing and/or stirring to second culture medium and first bent kind in the second culture dish for the material that is in the second culture dish bottom leaks, thereby makes the difficult condition that takes place oxygen inadequately, the high and suppression aspergillus niger growth of material bottom the second culture dish.
Optionally, in S9, the culturing of the accumulated material is sequentially divided into standing heat preservation culturing, intermittent ventilation culturing in the early growth stage of aspergillus niger, and continuous ventilation heat preservation culturing in the middle and later growth stages of aspergillus niger.
By adopting the technical scheme, the heat generated in the culture medium is gradually increased along with the growth of the aspergillus niger, and different ventilation modes are suitable for cooling the aspergillus niger at different growth stages, so that the culture medium is not easy to generate the condition of inhibiting the growth of the aspergillus niger due to overhigh temperature.
In summary, the present application has the following beneficial effects:
1. the method comprises the steps of putting materials into a kettle body from a material port, closing the material port by using a baffle plate, opening a first valve to enable water vapor to be introduced into the kettle body from a vapor inlet pipe, controlling a driving assembly to enable the driving assembly to drive the kettle body to rotate, enabling the kettle body to drive the materials in the kettle body to rotate, heating and sterilizing the materials, and gradually increasing the pressure in the kettle body due to the fact that air is introduced into the kettle body for a long time, so that the sterilization effect is further achieved, and the purpose of reducing the survival rate of the mixed bacteria in the materials is further achieved; in addition, the kettle body is rotationally connected with the rack, so that workers can conveniently rotate the attached drawings in the direction close to the ground, the material port faces the ground, and the materials in the kettle body can be poured out conveniently by the workers;
2. through the arrangement of the shovel plate, the shovel plate rotates and further stirs the materials in the kettle body, so that the heating uniformity of the materials in the kettle body is improved;
3. according to the method, the sterilization kettle is adopted, so that the sterilization effect on each culture medium is improved, and the effect of reducing the quantity of mixed bacteria in the culture medium is achieved; and Aspergillus niger is separated and screened for many times, so that the Aspergillus niger strain has the advantages of high purity, strong activity, low infection rate and high saccharification efficiency, and the culture method of the Aspergillus niger strain is simple to operate, low in equipment operation rate and capable of saving resources.
Drawings
FIG. 1 is a schematic view of the overall structure of a sterilization kettle according to the present application;
FIG. 2 is a partial cross-sectional view of the retort of the present application;
fig. 3 is an enlarged schematic view of a portion a in fig. 2.
Reference numerals: 100. a frame; 200. a kettle body; 210. a material port; 211. a striker plate; 230. a rotating shaft; 231. an accommodating chamber; 232. connecting holes; 240. a steam inlet pipe; 241. a first valve; 250. a steam exhaust pipe; 251. a second valve; 260. a cavity; 261. a vapor vent; 300. a drive assembly; 310. a drive motor; 320. a drive gear; 330. a transmission gear; 400. a shovel plate; 410. a positioning sleeve; 411. a baffle plate; 500. an adjustment assembly; 510. an adjusting gear; 520. adjusting the gear ring; 530. the gear is rotated.
Detailed Description
The present application is described in further detail below with reference to figures 1-3 and examples.
Aspergillus niger in the examples of the present application is Aspergillus niger UV _ (11-48).
The embodiment of the application discloses a sterilization kettle. Referring to fig. 1, the sterilization kettle includes a spherical kettle body 200 and a frame 100 for supporting the kettle body 200.
Referring to fig. 2, a material port 210 for loading and unloading is formed in the kettle body 200, and a material blocking plate 211 for closing the material port 210 is rotatably connected to the kettle body 200. In order to heat the materials in the kettle body 200, a steam inlet pipe 240 for supplying steam into the kettle body 200 is communicated with the kettle body 200, and in the embodiment, the steam in the steam inlet pipe 240 is provided by a boiler. The steam inlet pipe 240 is provided with a first valve 241 for adjusting the water vapor flux in the steam inlet pipe 240, the kettle body 200 is communicated with a steam exhaust pipe 250 for exhausting the gas in the kettle body 200, and the steam exhaust pipe 250 is provided with a second valve 250 for adjusting the water vapor flux in the steam exhaust pipe 250. When the materials in the kettle body 200 are heated, the first valve 241 is opened, so that the water vapor flows into the kettle body 200 through the steam inlet pipe 240, the second valve 241 is opened, the second valve 241 is closed after the water vapor extrudes the original air in the kettle body 200, and the materials in the kettle body 200 are heated and pressurized by the water vapor, so that the sterilization treatment of the materials is realized.
In order to improve the heating uniformity of the materials in the kettle body 200, the rotating shaft 230 is welded on the kettle body 200, and the rotating shaft 230 is rotatably connected with the rack 100, so that the kettle body 200 is rotatably connected with the rack 100, the material port 210 rotates towards the direction close to or far away from the ground, and the rotating kettle body 200 can stir the materials.
Referring to fig. 2 and 3, in order to further improve the heating uniformity of the material in the kettle 200, a cavity 260 is formed on the side wall of the kettle 200, a steam vent 261 communicated with the interior of the kettle 200 is formed on the side wall of the cavity 260, and the steam vent 261 is smaller than the particle size of the material. The two ends of the rotating shaft 230 are both provided with accommodating cavities 231, and the side walls of the accommodating cavities 231 are provided with a plurality of connecting holes 232 communicated with the cavity 260. One end of the rotating shaft 230 extends out of the kettle body 200 and is connected with the steam inlet pipe 240 through a rotary joint, so that the accommodating cavity 231 of the rotating shaft 230 is communicated with the steam inlet pipe 240, and the other end of the rotating shaft 230 extends out of the kettle body 200 and is connected with the steam exhaust pipe 250 through a rotary joint, so that the accommodating cavity 231 of the rotating shaft 230 is communicated with the steam exhaust pipe 250. The water vapor in the steam inlet pipe 240 enters the cavity 260 through the accommodating cavity 231 of the rotating shaft 230 and enters the kettle body 200 through the steam through hole 261, and the diffusion range of the water vapor is enlarged due to the arrangement of the cavity 260.
The frame 100 is provided with a driving assembly 300 for driving the kettle body 200 to rotate. The driving assembly 300 includes a driving motor 310, a transmission gear 330, and a driving gear 320. The driving motor 310 is fixedly connected with the frame 100 through bolts, the transmission gear 330 is coaxially welded with the rotating shaft 230, the driving gear 320 is coaxially welded with an output shaft of the driving motor 310, and the driving gear 320 is meshed with the transmission gear 330. The driving motor 310 is started, so that the driving motor 310 drives the driving gear 320 to rotate, and the driving gear 320 is meshed with the transmission gear 330, so that the transmission gear 330 drives the rotating shaft 230 to rotate the kettle body 200 under the action of the driving gear 320.
In order to further stir the materials in the kettle body 200, a shovel plate 400 is connected to the rotating shaft 230, and the shovel plate 400 is arranged along the axis of the rotating shaft 230. Shovel board 400 one side is connected with pivot 230 rotation, and the opposite side stretches into in the material. The cross section of the shovel plate 400 is arc-shaped, so that when the shovel plate 400 stirs the materials, the shovel plate can receive part of the materials after contacting the materials, so that the materials in the middle position are turned out, and the temperature uniformity of the materials in the kettle body 200 is further improved.
The shovel 400 is integrally formed with a positioning sleeve 410, the positioning sleeve 410 is rotatably sleeved on the outer side of the rotating shaft 230, so that the shovel 400 is rotatably connected with the rotating shaft 230, the rotating directions of the shovel 400 and the rotating shaft 230 are opposite, and the rotating shaft 230 is provided with an adjusting assembly 500 for adjusting the rotation of the shovel 400.
The adjusting assemblies 500 are provided in two sets, the two sets of adjusting assemblies 500 are symmetrically arranged with the central line of the rotating shaft 230 as the symmetry axis, and the two sets of adjusting assemblies are both arranged in the cavity 260. Adjustment assembly 500 includes an adjustment ring gear 520, a rotation gear 530, and an adjustment gear 510. The rotating gear 530 is coaxially welded with the rotating shaft 230, and the adjusting gear 510 is engaged with the rotating gear 530 and is rotatably connected with the sidewall of the cavity 260. The baffle 411 is welded on the positioning sleeve 410, and the adjusting gear ring 520 is sleeved outside the adjusting gear 510 and meshed with the adjusting gear 510. Baffle 411 is welded to adjusting ring gear 520, and adjusting ring gear 520 is disposed coaxially with locating sleeve 410 such that the axis of adjusting ring gear 520 is collinear with the axis of rotation of blade 400. The baffle 411 is arranged to guide the water vapor flowing out of the cavity 260, so as to further improve the diffusion range of the water vapor.
The implementation principle of a sterilization kettle in the embodiment is as follows: the materials to be sterilized are loaded into the kettle body 200, the material blocking plate 211 covers the material opening 210, and the first valve 241 is opened, so that the water vapor enters the kettle body 200 through the steam inlet pipe 240, and the temperature and the pressure in the kettle body 200 are increased.
The driving motor 310 is started, so that the driving motor 310 drives the driving gear 320 to rotate, the transmission gear 330 drives the rotating shaft 230 to rotate, and the kettle body 200 rotates, so as to stir the materials in the kettle body 200.
When the rotating shaft 230 rotates, the rotating gear 530 is driven to rotate, so that the adjusting gear 510 rotates to drive the adjusting gear ring 520 to rotate, so that the adjusting gear ring 520 drives the shovel plate 400 to rotate reversely, and further stirring of the materials in the kettle body 200 is realized.
Based on the sterilization kettle, the application also provides a culture method of the aspergillus niger, which comprises the following steps: s1, preparing a primary culture medium, a triangular flask culture medium, a box culture medium and a pond culture medium, wherein the primary culture medium is prepared by mixing the following substances in parts by weight: 1100 parts of distilled water 990-5, 2.5-3.5 parts of sodium nitrate, 0.5-1.5 parts of dipotassium hydrogen phosphate, 0.3-0.6 part of potassium chloride, 0.2-0.7 part of magnesium sulfate, 0.008-0.013 part of ferrous sulfate, 1.8-2.1 parts of agar and 27-32 parts of sucrose, and the prepared primary culture medium is loaded into a primary culture dish; uniformly mixing bran and water in a mass ratio of 1:0.5 to prepare a culture material, putting the culture material into a sterilization kettle for sterilization, putting the sterilized culture material into a first culture dish, and finally shaking the first culture dish to cool the culture material in the first culture dish to 30-35 ℃ to prepare the first culture medium; the second culture medium is prepared by uniformly mixing bran and water in a mass ratio of 1:0.5 to prepare a culture material, sterilizing the culture material by using a sterilization kettle, filling the sterilized culture material into a second culture dish, and cooling to 30-35 ℃ of the culture material; the third culture medium is prepared by uniformly mixing bran and rice hull with the mass ratio of 1:0.5:0.1-0.15 with water to prepare a first culture material, sterilizing the first culture material by using a sterilization kettle, loading the sterilized first culture material into a third culture dish, and cooling the first culture material to the temperature of 30-35 ℃.
S2, primary culture and separation of Aspergillus niger: after culturing the aspergillus niger in a primary culture medium, separating qualified aspergillus niger in a sterile environment;
s3, inoculating qualified Aspergillus niger separated in S2 into the first culture medium, and shaking the first culture dish to uniformly mix the Aspergillus niger and the first culture medium;
s4, placing a first culture dish containing Aspergillus niger and a first culture medium in an environment of 32-33 ℃, performing heat preservation culture for 2.5-4 days, turning over the first culture dish for multiple times, placing the first culture dish in the environment of 32-33 ℃, continuing to perform heat preservation culture for 6-9 days, and finally separating qualified Aspergillus niger in a sterile environment to obtain a first yeast strain;
s5, inoculating the first koji seeds into a second culture medium, uniformly mixing the first koji seeds with the second culture medium, stacking the first koji seeds and the second culture medium in a second culture dish, and then putting the second culture dish containing the first koji seeds and the second culture medium into an environment at 28-33 ℃ for culturing for 4-6 hours;
s6, spreading a mixture of the first koji species and the second culture medium in the second culture dish to a thickness of 2.5-3.5cm, then continuously culturing for 30h at 28-33 ℃ to obtain a prefabricated koji, wherein when the second culture medium and the first koji species are cultured for 10-12h, the second culture dish is inverted up and down for 1 time, when the culture is continued for 8-9h, the mixture of the first koji species and the first culture medium is subjected to one-time material turning treatment, after the culture is continued for 5-6h, the second culture dish is inverted up and down for 1 time, and the culture is continued;
s7, drying the prefabricated seed koji for 4-6 hours, and then separating qualified Aspergillus niger in a sterile environment to obtain the seed koji;
s8, inoculating the seed starter to a third culture medium for culture, uniformly mixing the seed starter and the third culture medium in a third culture dish, stacking the seed starter and the third culture medium together, and then placing the third culture dish in an environment with the temperature of 28-33 ℃ for culture for 4-6h to obtain a stacking material;
s9, spreading the stacked materials in a third culture dish to a thickness of 25-30cm, and adjusting the ambient temperature to 28-33 ℃ for culturing for 28-34h to obtain the bran koji.
In order to adapt to the production conditions of the aspergillus niger at different stages, the culture of the accumulated materials is sequentially divided into standing heat preservation culture, intermittent ventilation culture in the early growth stage of the aspergillus niger and continuous ventilation heat preservation culture in the middle and later growth stages of the aspergillus niger, and the ventilation adopts mechanical blowing of an air blower.
The present embodiment is only for explaining the present application, and it is not limited to the present application, and those skilled in the art can make modifications of the present embodiment without inventive contribution as needed after reading the present specification, but all of them are protected by patent law within the scope of the claims of the present application.
Claims (10)
1. A sterilization kettle is characterized by comprising a rack (100) and a kettle body (200), wherein a material port (210) for charging and discharging is formed in the kettle body (200), a material baffle plate (211) for closing the material port (210) is arranged on the kettle body (200), the kettle body (200) is rotatably connected with the rack (100) so that the material port (210) rotates towards a direction close to or far away from the ground, a steam inlet pipe (240) for supplying steam into the kettle body (200) is communicated with the kettle body (200), a first valve (241) for adjusting the flux of the steam in the steam inlet pipe (240) is arranged on the steam inlet pipe (240), a steam exhaust pipe (250) for exhausting the gas in the kettle body (200) is communicated with the kettle body (200), and a second valve (251) for adjusting the flux of the steam in the steam exhaust pipe (250) is arranged on the steam exhaust pipe (250), the frame (100) is provided with a driving assembly (300) for driving the kettle body (200) to rotate.
2. The autoclave of claim 1, wherein: fixedly connected with pivot (230) is gone up in cauldron body (200), and pivot (230) rotate with frame (100) and be connected, cavity (260) have been seted up on cauldron body (200) lateral wall, and set up on cauldron body (200) and pass through steam hole (261) with cavity (260) intercommunication, pivot (230) both ends have all been seted up and have been held chamber (231), hold chamber (231) lateral wall and last set up a plurality of connecting hole (232) that communicate with cavity (260), pivot (230) one end is stretched out cauldron body (200) and is connected with admission pipe (240) through rotary joint to make holding chamber (231) and admission pipe (240) intercommunication of pivot (230), pivot (230) other end stretches out cauldron body (200) and is connected with steam extraction pipe (250) through rotary joint, so that the chamber (231) that holds of pivot (230) communicate with steam extraction pipe (250).
3. The autoclave of claim 2, wherein: the driving assembly (300) comprises a driving motor (310), a transmission gear (330) and a driving gear (320), the driving motor (310) is fixedly connected with the rack (100), the transmission gear (330) is coaxially and fixedly connected with the rotating shaft (230), the driving gear (320) is coaxially and fixedly connected with an output shaft of the driving motor (310), and the driving gear (320) is meshed with the transmission gear (330).
4. The autoclave of claim 3, wherein: the rotating shaft (230) is connected with a shovel plate (400) and is arranged along the axis of the rotating shaft (230).
5. The autoclave of claim 4, wherein: the shovel plate (400) is rotatably connected with the rotating shaft (230), the rotating directions of the shovel plate (400) and the rotating shaft (230) are opposite, and an adjusting assembly (500) used for adjusting the rotation of the shovel plate (400) is arranged on the rotating shaft (230).
6. The autoclave of claim 5, wherein: adjust subassembly (500) including adjusting ring gear (520), rotating gear (530) and adjusting gear (510), rotating gear (530) and pivot (230) coaxial fixed connection, adjusting gear (510) and rotating gear (530) meshing to rotate with cavity (260) lateral wall and be connected, adjust ring gear (520) cover and establish in the adjusting gear (510) outside, and mesh with adjusting gear (510), adjust ring gear (520) and shovel board (400) fixed connection, and the axis of adjusting ring gear (520) and the rotation axis collineation setting of shovel board (400).
7. A method for culturing Aspergillus niger is characterized in that: the method comprises the following steps: s1, preparing a primary culture medium loaded in a primary culture dish, a first culture medium loaded in a first culture dish, a second culture medium loaded in a second culture dish and a third culture medium loaded in a third culture dish, wherein the first culture medium, the second culture medium and the third culture medium are sterilized by a sterilization kettle during preparation;
s2, primary culture and separation of Aspergillus niger: after culturing the aspergillus niger in a primary culture medium, separating qualified aspergillus niger in a sterile environment;
s3, inoculating qualified Aspergillus niger separated in S2 into the first culture medium, and shaking the first culture dish to uniformly mix the Aspergillus niger and the first culture medium;
s4, placing a first culture dish containing Aspergillus niger and a first culture medium in an environment of 32-33 ℃, after heat preservation culture, turning over the first culture dish for multiple times, placing the first culture dish in the environment of 32-33 ℃ for continuous heat preservation culture, and finally separating qualified Aspergillus niger in a sterile environment to obtain a first yeast strain;
s5, inoculating the first koji species into a second culture medium, uniformly mixing the first koji species with the second culture medium, stacking the first koji species and the second culture medium in a second culture dish, and then putting the second culture dish containing the first koji species and the second culture medium into an environment at 28-33 ℃ for culture;
s6, spreading the mixture of the first koji in the second culture dish and the second culture medium to the thickness of the mixture of 2.5-3.5cm, and continuously culturing at 28-33 ℃ to obtain the prefabricated koji;
s7, drying the prefabricated seed koji, and separating qualified Aspergillus niger in an aseptic environment to obtain the seed koji;
s8, inoculating the seed starter to a third culture medium for culture, uniformly mixing the seed starter and the third culture medium in a third culture dish, stacking the seed starter and the third culture medium together, and then placing the third culture dish in an environment with the temperature of 28-33 ℃ for culture to obtain a stacking material;
s9, spreading the piled material prepared in the S8 in a third culture dish, wherein the spread thickness is 25-30cm, and placing the third culture dish loaded with the piled material in an environment with the temperature of 28-33 ℃ for culture to obtain the bran koji.
8. The method for culturing Aspergillus niger according to claim 7, wherein: in S1, the primary culture medium is prepared by mixing the following substances in parts by weight: 1100 parts of distilled water 990-type-; uniformly mixing bran and water in a mass ratio of 1:0.5 to prepare a culture material, putting the culture material into a sterilization kettle for sterilization, putting the sterilized culture material into a first culture dish, and finally shaking the first culture dish to cool the culture material in the first culture dish to 30-35 ℃ to prepare the first culture medium; the second culture medium is prepared by uniformly mixing bran and water in a mass ratio of 1:0.5 to prepare a culture material, sterilizing the culture material by using a sterilization kettle, filling the sterilized culture material into a second culture dish, and cooling to 30-35 ℃ of the culture material; the third culture medium is prepared by uniformly mixing bran and rice hull with the mass ratio of 1:0.5:0.1-0.15 with water to prepare a first culture material, sterilizing the first culture material by using a sterilization kettle, loading the sterilized first culture material into a third culture dish, and cooling the first culture material to the temperature of 30-35 ℃.
9. The method for culturing Aspergillus niger according to claim 7, wherein: in S6, in the first koji culture process, the material in the second petri dish needs to be turned over and/or the second petri dish needs to be turned over.
10. The method for culturing Aspergillus niger according to claim 7, wherein: in S9, the culture of the accumulated materials is divided into standing heat preservation culture, intermittent ventilation culture in the early growth stage of Aspergillus niger and continuous ventilation heat preservation culture in the middle and later growth stages of Aspergillus niger.
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