WO2017020853A1 - 一种固态生物反应装置、使用方法及其应用 - Google Patents

一种固态生物反应装置、使用方法及其应用 Download PDF

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Publication number
WO2017020853A1
WO2017020853A1 PCT/CN2016/093267 CN2016093267W WO2017020853A1 WO 2017020853 A1 WO2017020853 A1 WO 2017020853A1 CN 2016093267 W CN2016093267 W CN 2016093267W WO 2017020853 A1 WO2017020853 A1 WO 2017020853A1
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Prior art keywords
main tank
tank body
culture
disposed
hole
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PCT/CN2016/093267
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English (en)
French (fr)
Inventor
姜文侠
张笑然
马延和
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中国科学院天津工业生物技术研究所
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Priority to EP16832341.8A priority Critical patent/EP3333252B1/en
Priority to US15/750,126 priority patent/US10851334B2/en
Publication of WO2017020853A1 publication Critical patent/WO2017020853A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/16—Solid state fermenters, e.g. for koji production
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/04—Apparatus for enzymology or microbiology with gas introduction means
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/16—Apparatus for enzymology or microbiology containing, or adapted to contain, solid media
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/48—Holding appliances; Racks; Supports
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/50—Means for positioning or orientating the apparatus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/06—Plates; Walls; Drawers; Multilayer plates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
    • C12M27/10—Rotating vessel
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/04—Filters; Permeable or porous membranes or plates, e.g. dialysis
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
    • C12M41/18—Heat exchange systems, e.g. heat jackets or outer envelopes
    • C12M41/22—Heat exchange systems, e.g. heat jackets or outer envelopes in contact with the bioreactor walls
    • 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
    • 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
    • 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
    • C12N1/16—Yeasts; Culture media therefor
    • C12N1/18—Baker's yeast; Brewer's yeast

Definitions

  • the invention belongs to the field of bioengineering, and particularly relates to a solid biological reaction device, a using method and an application thereof.
  • Filamentous bacteria are the production bacteria of many fermented products, and are widely used in industries such as antibiotics, organic acids, enzyme preparations and biological pesticides. Spore inoculation is a commonly used inoculation method for filamentous fungi fermentation. In fermentation production, the spores of filamentous fungi are generally prepared by surface culture (such as solid culture). For example, the citric acid industry generally adopts a triangular flask for static solid bottle culture.
  • the key to the large-scale preparation of filamentous spores is to develop equipment that meets the requirements of the process.
  • the domestic equipment for the large-scale preparation of pure spores of filamentous fungi is still blank, and there are few companies that manufacture and sell such devices internationally. .
  • Currently, only the VB Spore Box of the Vogelbusch Biocommodities GmbH is seen on the market.
  • the device uses more The technique of layer shallow plate culture, vacuum collection of pure spores, spore culture, drying and collection are carried out in a closed system with a production capacity of 0.8-1.4 kg of Aspergillus niger spores per batch for 14 days.
  • the sterilizing agent since the device is sterilized by chemical fumigation, the sterilizing agent must be drained so as not to affect the growth of the microorganisms, and the discharged residual sterilizing agent needs to be neutralized to prevent environmental pollution; the agar medium is sterilized separately and then sterilized.
  • the risk of infection in the transfer process is high; the cleaning and preparation time is long, the manual labor is still large; the cost of the agar medium is also higher; in addition, the way of collecting the spores under negative pressure will also be carried Come to a higher risk of infection.
  • CN 104630020 A discloses a solid-state biological reaction device for large-scale preparation of filamentous spores, the dried spore powder prepared by the device can be used for fermenting inoculation immediately, and can be stored in spores for a long time. Collected in the bottle.
  • the structure is simpler, more compact, lower in manufacturing cost, and more flexible filaments.
  • Microspore preparation device in particular, the agitation system of the device can greatly reduce the manufacturing cost of the device if the design is simplified.
  • CN 203559041 U discloses a movable wine fermenter, which is suitable for wine fermentation, whose movement function is horizontal movement, can not achieve the rotation of the tank posture, and does not have the function of rotary mixing, and thus is not suitable for solid biological reaction.
  • CN 102924146 A discloses a movable drum compost reactor, which is designed for use in a solid-state bioreactor. The device realizes rotary mixing by means of a drum, but its movement mode is limited to horizontal movement, and the rotation mode is limited to fixed. The rotation of the axis does not allow for a change in the attitude of the container space, nor is it suitable for the preparation of purely cultured filamentous spore species by solid biological reaction.
  • the invention provides a solid-state biological reaction device, a use method and an application thereof, the purpose of which is to achieve low-intensity mixing of materials through the change of the space attitude and the rotation of the container in the small solid-state reaction device, and at the same time satisfy the culture and materials. Mixing different requirements for the attitude of the device, and simplifying the design, saving costs.
  • the invention can be used not only for the large-scale preparation of filamentous spores, but also for general microbial solid state culture and solid state fermentation.
  • the present invention provides a solid state bioreactor comprising a main can body, the device further comprising a support supporting the main can body, the support being capable of changing the posture of the main can body such that the main can body is in a horizontal position Realize the rotation and realize the static culture in the vertical position;
  • the main tank body when the main tank body is in the horizontal position, the main tank body can be manually moved or automatically driven by the driving device; when the main tank body is in the vertical position, the main tank body is statically cultured.
  • the solid-state biological reaction device of the invention realizes the free-spinning mixing of the tank body in two different postures of upright and horizontal, and the main tank body is placed in a horizontal state, the tank body can be rotated, and the rotation speed can be controlled.
  • the material in the tank is thoroughly mixed with the inserted bacteria, and can be rotated and mixed periodically in the subsequent cultivation process to uniformly grow the culture without stirring the paddles, the shaft seal and the like, thereby avoiding stirring the tank by stirring the slurry or the like. The material thus destroys the culture.
  • a center of the top end of the main tank body is provided with a rotary shaft
  • the bracket includes a beam for connecting to a rotary shaft of the main tank;
  • the bracket is provided with a support for supporting the main can body.
  • the main tank body is provided with at least one sieve plate for maintaining the material position
  • the sieve plate has an open cell ratio of from 1 to 90%, preferably from 1 to 60%.
  • the outer edge of the sieve plate is sealingly connected to the inner wall of the main tank body, and the sealing connection may be a flexible seal or a rigid seal.
  • the sieve plate may not be a fully-opened sieve plate, and the annular non-aperture zone can maintain a part of the free liquid medium in the side wall during the high-temperature sterilization process, thereby ensuring that the liquid medium can be cultured with the medium particles.
  • Fully absorb the swelling ensure that the seed liquid is in full contact with the culture medium and fully absorbed during the mixing process after inoculation.
  • the ratio of the diameter of the opening area of the sieve plate to the outer diameter of the sieve plate is 0.1-1, and may be, for example, 0.1, 0.2, 0.3, 0.4, 0.5, 055, 0.6, 0.7, 0.8, 0.9 or 1. It is preferably from 0.5 to 0.9.
  • the opening on the sieve plate may be, but not limited to, a circular hole and/or an oblong hole groove, and may also be an oval hole, a triangular hole, a polygonal hole or an irregular hole.
  • the circular hole has a diameter of 0.2-20 mm, and may be, for example, 0.2 mm, 0.3 mm, 0.4 mm, 0.5 mm, 0.6 mm, 0.8 mm, 1 mm, 2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 8 mm, 10 mm. 12mm, 15mm, 16mm, 18mm, 19mm or 20mm, preferably 0.5-3mm.
  • the oblong hole groove has a width of 0.2-20 mm, and may be, for example, 0.2 mm, 0.3 mm, 0.4 mm, 0.5 mm, 0.6 mm, 0.8 mm, 1 mm, 2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 8 mm, 10 mm, 12 mm, 15 mm, 16 mm, 18 mm, 19 mm or 20 mm, preferably 0.5-3 mm.
  • the opening ratio of the sieve plate can be opened according to the requirement of the ventilation amount of the strain, and the opening ratio and the pore size are adjusted.
  • the main tank body comprises an end cover, a cylinder body and a lower head from top to bottom;
  • the end cover includes a rotary shaft with a pin hole disposed at a center of the top end of the end cover, and an exhaust gas filter, a liquid filter and an inoculating port disposed on the end cover;
  • the cylinder includes a sight glass disposed on the inner wall of the cylinder, and a temperature sensor disposed at a lower portion of the cylinder and a temperature sensor sleeve disposed outside the temperature sensor;
  • the lower head includes a discharge valve disposed at a middle portion of a bottom end of the lower head, and a feed disposed on the lower head Gas filter.
  • the sight glass is circular or strip-shaped, preferably strip-shaped.
  • a check valve is disposed between the intake filter and the lower head.
  • a pipe joint is provided upstream of the intake filter for connecting the compressed air line.
  • the exhaust gas filter, the liquid filter and the inlet filter have a pore diameter of 0.1 to 0.22 ⁇ m, for example 0.1 ⁇ m, 0.15 ⁇ m, 0.2 ⁇ m, 0.21 ⁇ m or 0.22 ⁇ m, preferably 0.2 ⁇ m.
  • the main tank body comprises an upper screen plate disposed at an interface between the end cover and the inner portion of the cylinder body, and a lower screen plate disposed at an interface between the barrel body and the inner side of the lower head.
  • the device may be equipped with one set of sieve plates including one upper sieve plate and one lower sieve plate, or multiple sets of holes having different opening diameters and different shapes and different geometric sizes. And sieve plates with different opening ratios, suitable for different strains and materials.
  • an upper limit ring is disposed on the upper edge of the cylinder, and a lower limit ring is disposed on the upper edge of the lower head.
  • the outer portion of the cylinder is provided with a jacket, and preferably the outer portion of the cylinder and the lower head is provided with a jacket, and the jacket comprises an inlet pipe disposed at a lower portion of the jacket and an outlet disposed at an upper portion of the jacket Water pipe.
  • the jacket may cover all of the bare parts of the barrel and the lower head without components, or may cover only the barrel portion or may cover only the lower head.
  • an exhaust gas regulating valve is disposed between the end cover and the exhaust gas filter.
  • an isolation valve is disposed between the end cap and the liquid filter.
  • the bracket can change the attitude of the main tank, which can be a carrying vehicle, preferably a lever type trailer;
  • the lever type trailer includes a base and a column, the base of the base and the column is provided with a wheel, the base and the column are integrally rotated by a rotation axis of the wheel, and the base and the column rotate with the axle, the axle It can be a single axle or two axles;
  • the base includes a vertical support leg and an annular support disposed above the base, the annular support Supporting the main tank body;
  • the post includes a horizontal support leg and a beam attached to the horizontal support leg for attachment to the main can body.
  • the inside of the column is provided with a rotary driving device for driving the main can body to rotate.
  • the rotary drive device can be electrically driven or pneumatically operated, and the rotational speed can be adjusted.
  • the horizontal support leg and the vertical support leg may also be casters with a locking device.
  • the horizontal support legs and the vertical support legs can be adjusted by threads, and the stability of the main can body can be ensured in an upright or horizontal position.
  • the main tank body is connected with the lever type trailer, and the horizontal and vertical positions can be changed not only by the main tank body but also by the lever type trailer, so that the material and the bacterial suspension can be mixed; meanwhile, the main tank can be driven by the lever type trailer.
  • the body is easily moved to the desired position and moved, without the need for additional steps, reducing the possibility of contamination.
  • the upper surface of the annular seat is provided with a sliding bearing.
  • the inner side of the column is provided with a supporting wheel for supporting the side wall of the main tank horizontally and horizontally.
  • the supporting roller when the device is placed in a horizontal position, the supporting roller is used to carry the weight of the main tank body, and at the same time, the main tank body can be freely rotated, and the height of the supporting wheel can be adjusted by bolts on the supporting wheel seat, so that The tank wall is closely attached and can be rotated 360°.
  • the beam is opened with a vertical through hole, and a through hole is fixed in the through hole to fix a rotating shaft on the main can body.
  • the beam has a horizontal threaded hole perpendicularly intersecting the through hole for mounting a fastening screw to secure the sleeve.
  • the sleeve is provided with a pin hole 1 and a pin hole 2 for mounting a fixing pin.
  • the inner surface of the lower end of the sleeve is provided with a sliding bearing.
  • the main tank body and the support material are made of materials capable of withstanding steam sterilization at 121 ° C and 0.1 MPa, and may be selected from, but not limited to, stainless steel, carbon steel, non-ferrous metals, light alloys, plastics, ceramics, Any one or a mixture of at least two of glass or glass.
  • the end cap is elliptical, spherical, spherical, dished or flat, preferably elliptical or dish shaped, further preferably elliptical.
  • the lower head is elliptical, spherical, spherical, dished or conical, preferably tapered.
  • the present invention provides a method of solid-culturing a microorganism, using the solid biological reaction device of the first aspect, the method comprising the steps of:
  • mixing is carried out multiple times during the cultivation
  • the culture when the strain for the fermenter is cultured, the culture is transferred to the fermenter after the completion of the culture, and the culture is expanded.
  • the specific steps of transplanting the fermenter include: opening the isolation valve and the exhaust gas regulating valve, adding water or an aqueous solution to the tank by means of filtering sterilization, and then closing the isolation valve and the exhaust gas regulating valve to place the main tank body Pour, continue the operation of the rotary mixing until most of the bacteria have been washed through the sight glass. Stand up the main tank, unscrew the fastening screw, and insert a pin into the threaded hole of the beam to suspend the entire main tank in a higher position.
  • the transfer tube is connected between the discharge valve and the top of the fermentation tank, and the steam is fed through the valve to a temperature of not less than 121 ° C and a pressure of not less than 0.1 MPa (gauge pressure).
  • the pipeline was sterilized by less than 20 minutes.
  • the seeding method can also be carried out according to the following steps: after the installation is completed After the connecting pipe is sterilized, the exhaust gas regulating valve and the isolation valve are closed, the main tank bottom valve and the top of the fermenting tank are opened, so that the main tank body communicates with the upper part of the fermenting tank, and then the tank pressure of the fermenting tank is lowered and utilized. The pressure difference moves the strain suspension into the fermentor. The water can be added several times repeatedly to move the bacteria into the fermenter as thoroughly as possible. After the seeding is completed, the top of the fermenter is moved to the seeding valve, and the transplanting pipe is removed.
  • step (1) specifically comprises the following steps:
  • step (2) specifically comprises the following steps:
  • compressed air may be introduced from the air tube joint to blow the culture.
  • the invention provides a solid bioreactor according to the first aspect, for use in a culture microorganism or solid state fermentation using the method as described in the second aspect.
  • the reaction device of the invention can be used not only for large-scale preparation of filamentous spores, but also for general microbial solid state culture and solid state fermentation.
  • the microorganism may be selected from, but not limited to, a filamentous fungus, a yeast, or a bacterium.
  • the present invention has the following beneficial effects:
  • the device of the invention is relatively simple, in particular, the mixing of materials adopts the combination of the vehicle tank body and the rotary body, and realizes the free-slewing mixing of the tank body in two different postures of upright and horizontal;
  • the sieve plate of the device of the invention adopts a non-full opening sieve plate, which can fix the position of materials and bacteria when the main tank body is in a horizontal position, and restrict the movement of materials and bacteria, so that the mixing of materials and bacteria is more uniform;
  • the device of the present invention can be used for conveniently solid-culturing microorganisms, especially filamentous spores and various single-cell microorganisms, and the prepared spores can be conveniently and aseptically transferred to the fermentor in the form of a suspension. It can be dried together with the culture medium and stored in the device for a long time.
  • the preparation and transfer process of the spores is much simpler than the bran; the prepared single-cell microorganisms can also be conveniently and aseptically in the form of a bacterial suspension. Transfer to the fermenter.
  • the device of the invention carries out charging, inoculation, cultivation, adding water and transplanting work in an upright posture, and completes the sterilization and material rotation mixing work in a horizontal posture, which is not only quick to use, convenient to move, but also saves occupation. A large number of manufacturing costs of the mixing system, low maintenance costs.
  • Figure 1 is a schematic view of the main tank body of the present invention
  • Figure 2 is a front view of the vehicle of the present invention.
  • Figure 3 is a left side view of the vehicle of the present invention.
  • Figure 4 is a plan view of the vehicle of the present invention.
  • Figure 5 is a schematic view showing the inoculation/culture state of the present invention.
  • Figure 6 is a schematic view showing the sterilization/swing mixing state of the present invention.
  • Figure 7 is a schematic view showing the state of seeding of the present invention.
  • Figure 8 is a schematic view of a four-valve shifting device of the present invention.
  • Figure 9 is a schematic view of the sieve plate of the present invention and its opening area
  • 1-main can body 3-end cap; 4-tank; 5-lower head; 6-tail gas filter; 7-liquid filter; 8-inoculation port; 9-view mirror; Temperature sensor; 11-temperature sensor casing; 12-jacket; 13-inlet water inlet pipe; 14-circulating water outlet pipe; 15-intake filter; 16-discharge valve; 17-check valve; 18-rotary shaft; 19-pin hole; 20-upper limit pull ring; 21-lower limit pull ring; 22-upper sieve plate; 23-lower sieve plate; 24-pipe joint; 25-tail gas regulating valve; 26-isolation valve; 27-screen plate; 28-axle; 29-base; 30-ring bearing; 31-column; 32-beam; 33-horizontal support leg; 34-vertical support leg; 35-vertical through hole; 36-set Cartridge; 37-pin hole 1; 38-pin hole 2; 39-wheel; 40-bearing wheel; 41-threaded hole; 42-sliding bearing 1; 43
  • Example 1 A solid state bioreactor
  • FIG. 1 it is a schematic diagram of a solid biological reaction device according to an embodiment of the present invention.
  • the main tank body 1 and the lever type trailer 2 are arranged, and the lever type trailer 2 rotates the main tank body 1 in a horizontal position and is statically cultured in a vertical position;
  • the main tank body 1 when the main tank body 1 is in the horizontal position, the main tank body 1 can be automatically driven by the manual dialing or driving device 49; when the main tank body 1 is in the vertical position, the main tank body 1 is statically cultured, such as Figure 6 shows.
  • the main tank body 1 includes an end cover 3, a cylinder 4 and a lower head 5 from top to bottom;
  • the end cap 3 includes a rotary shaft 18 having a pin hole 19 disposed at the center of the top end of the end cap 3, and an exhaust gas filter 6, a liquid filter 7, and an inoculating port 8 disposed on the end cap 3;
  • An exhaust gas regulating valve 25 is disposed between the exhaust filter 3 and the exhaust gas filter 6;
  • an isolation valve 26 is disposed between the end cap 3 and the liquid filter 7.
  • the cylinder 4 includes a sight glass 9 disposed inside the cylinder 4, and a temperature sensor 10 disposed at a lower portion of the cylinder 4 and a temperature sensor sleeve 11 disposed outside the temperature sensor 10; the upper edge of the cylinder 4 is disposed There is an upper limit ring 20.
  • the lower head 5 includes a discharge valve 16 disposed at the center of the bottom end of the lower head 5, and an intake filter 15 disposed on the lower head 5; the intake filter 15 and the lower head 5 A check valve 17 is disposed therebetween; a pipe joint 24 is disposed upstream of the intake filter 15 for connecting a compressed air line; and a lower limit ring 21 is disposed on the upper edge of the lower head 5.
  • the main tank body 1 is internally provided with an upper sieve plate 22 at the boundary between the end cover 3 and the inner portion of the cylindrical body 4, and a lower sieve plate 23 disposed at the inner boundary of the cylindrical body 4 and the lower sealing head 5 for maintaining the material position;
  • the sieve plate 27 has an opening ratio of 1-90%.
  • the lever type trailer includes a base 29 and a column 31.
  • the base of the base 29 and the column 31 is provided with an axle 28.
  • the base 29 and the column 31 rotate with the axle 28, and the wheel axle 28 is provided with wheels 39 at both ends. ;
  • the base 29 includes a vertical support leg 34 and an annular support 30 disposed above the base 29, the annular support 30 is for supporting the main can body 1; the upper surface of the annular support 30 is provided with a sliding Bearing 42.
  • the column 31 includes a horizontal support leg 33, and a beam 32 connected to the horizontal support leg 33 for connection to the main can body 4, and a support wheel 40 disposed on the right side of the horizontal support leg 33 for the main canister
  • the body 1 is horizontally placed and horizontally supported by the side walls.
  • the beam 32 is provided with a vertical through hole 35 through which a sleeve 36 for fixing the rotary shaft 18 of the main can body 1 is bored; the beam 32 has a through hole 35 Vertically intersecting horizontal threaded holes 41 for mounting fastening screws to secure the sleeve 36; the sleeve 36 having a pin hole 1 (37) and a pin hole 2 (38) for mounting a fixing pin; The inner surface of the lower end of the sleeve 36 is provided with a sliding bearing 43.
  • the method of using the device includes the following steps:
  • the fastening screw is screwed into the beam 32 through the threaded hole 41 and tightened to withstand the sleeve 36, and the sleeve 36 and the beam 32 are fastened into one body; the main tank body 1
  • the exhaust gas regulating valve 25 and the isolation valve 26 are opened to perform sterilization.
  • a compressed air tube is inserted at the pipe joint 24, and compressed air is blown to dry the intake filter 15, the exhaust gas filter 6, and the liquid filter 7, while the culture medium is cooled, and the isolation valve 26 is closed.
  • the tail gas regulating valve 25 is opened, and under the flame protection, the liquid seed of the microorganism to be cultured is connected from the inoculation port 8; after inoculation, the main tank body 1 is placed horizontally with the lever type trailer 2, and the pin of the pin hole 38 is pulled out.
  • the main tank body 1 is rotated by hand to rotate the material in the tank and the liquid seed to be mixed; after mixing, the pin is reinserted, the main tank body 1 is erected, compressed air is introduced for cultivation, and the water is passed through the water inlet pipe. 13 and the outlet pipe 14 is introduced into the jacket 12 with the same constant temperature water as the culture temperature to maintain the temperature of the culture; during the cultivation process, the main tank body 1 is intermittently laid down, and the operation of the rotary mixing is repeated several times;
  • the specific steps of the seeding include: opening the isolation valve 25 and the exhaust gas regulating valve 26, and adding water or an aqueous solution to the tank by means of filtering sterilization. Thereafter, the exhaust gas regulating valve 25 and the isolation valve 26 are closed, the main tank body 1 is lowered, and the operation of the rotary mixing is continued until most of the bacteria have been washed through the sight glass 9. The main can body 1 is raised, the fastening screw is unscrewed, and a pin is inserted into the threaded hole of the beam to suspend the entire main can body 1 at a higher position.
  • the transfer tube 45 is connected between the discharge valve 16 and the fermenter top inoculation valve 44, and the steam having a temperature of not less than 121 ° C and a pressure of not less than 0.1 MPa (gauge pressure) is introduced into the valve 46 and 47.
  • the tube is sterilized by maintaining it for at least 20 minutes.
  • a compressed air tube is connected at the pipe joint 24, and compressed air having a pressure higher than that of the fermenter 48 is introduced, and the bacteria in the device are suspended and pressurized into the fermentor 48. The water may be added several times repeatedly to move the bacteria into the fermentor 48 as thoroughly as possible.
  • the top tank valve 44 is closed and the seeding pipe is removed.
  • the seeding method can also be carried out according to the following steps: after the seeding tube 45 is installed and sterilized, the exhaust gas regulating valve 25 and the isolation valve 26 are closed, and the discharge valve 16 and the fermentation tank top valve 44 at the lower portion of the main tank body 1 are opened.
  • the main tank 1 is in air communication with the upper portion of the fermentor 48, after which the tank pressure of the fermentor 48 is lowered, and the bacterial suspension is transferred to the fermentor 48 by a pressure difference.
  • the sieve plate 27 may also be provided with a plurality of layers distributed in the main tank body 1.
  • the ratio of the diameter of the opening area of the sieve plate 27 to the outer diameter of the sieve plate 27 is 0.1-1, preferably 0.5- 0.9; the opening on the sieve plate 27 is a circular hole and/or an oblong hole; the diameter of the circular hole is 0.5-20 mm, preferably 1-10 mm; the width of the long circular groove is 0.5-20 mm, It is preferably from 1 to 10 mm.
  • the sight glass 9 is circular or strip-shaped, preferably strip-shaped.
  • the exhaust gas filter 6, the liquid filter 7, and the intake filter 15 have a pore diameter of 0.2 to 0.22 ⁇ m, preferably 0.2 ⁇ m.
  • the outer portion of the cylinder 4 is provided with a jacket 12, and preferably the outer portion of the cylinder 4 and the lower head 5 is provided with a jacket 12, and the jacket 12 is disposed at a lower portion of the jacket 12.
  • the inlet pipe 13 and the outlet pipe 14 disposed at the upper portion of the jacket 12.
  • the horizontal support leg 33 and the vertical support leg 34 may also be casters with a height adjustment device and a locking device.
  • the main tank body 1 and the carrier 2 are made of a material resistant to 121 ° C, 0.1 MPa steam, and may be selected from, but not limited to, stainless steel, carbon steel, non-ferrous metals, light alloys, Any one or a mixture of at least two of plastic, ceramic, enamel glass or glass.
  • the end cap 3 is elliptical, spherical, spherical, dished or flat, preferably elliptical or dish-shaped, further preferably elliptical.
  • the lower head 5 is elliptical, spherical, spherical, dished or tapered, preferably tapered.
  • the spore culture of Aspergillus niger was carried out by the apparatus described in Example 1, and the culture method was the same as that of the culture example of Example 1.
  • the composition of the aqueous medium solution is shown in Table 1, wherein the steps in the culture process are as follows:
  • the main tank body 1 has a full volume of 30L and is made of 316L stainless steel.
  • the end cap 3 is elliptical.
  • the lower head 5 has a cone shape with a cone angle of 120°.
  • the jacket covers the barrel 4 and the lower head 5. ;
  • the sieve plate 27 may also be provided with a plurality of layers distributed in the main tank body 1.
  • the opening ratio of the sieve plate 27 is 29%, and the ratio of the diameter of the opening portion to the outer diameter of the sieve plate 27 is 0.86;
  • the opening is a round hole
  • the diameter of the circular hole is 3 mm.
  • Corn kernel particles with a particle size ranging from 5 to 10 mm are placed in an excess of aqueous medium.
  • the composition of the aqueous medium is shown in Table 1.
  • the soaked corn cob particles are removed from the aqueous solution and placed on a lower sieve. Flat, the thickness of the layer is 10cm, and immersed for 12h at room temperature.
  • the culture method is the same as the culture step in Example 1. The specific parameters are as follows: constant temperature circulating water of 35 °C is continuously introduced into the jacket, and the culture temperature is maintained at 35 °C, and the air flow rate is 0.5.
  • the specific steps of the seeding process are the same as those in the first embodiment.
  • the specific parameters are as follows: when the main tank body 1 is placed in a horizontal state, the rotation speed is 10 rpm, and the rotation is continued for 10 minutes.
  • Example 2 For the spore culture of Aspergillus sp., the apparatus described in Example 1 was used, and the culture method was the same as that in Example 1.
  • the aqueous medium solution is shown in Table 2, wherein the steps in the culture process are as follows:
  • the main tank body 1 has a full volume of 45 L, the material is 304 stainless steel, the end cover 3 has a dish shape, the lower head 5 is spherical, and the jacket covers the barrel 4 and the lower head 5 with all bare parts;
  • the sieve plate 27 may also be provided with a plurality of layers distributed in the main tank body 1.
  • the upper sieve plate 22 has an opening ratio of 39%, the sieve hole is a circular hole, the diameter is 2 mm, and the ratio of the diameter of the opening portion to the outer diameter of the sieve plate 27 The ratio is 0.79, the lower sieve plate opening ratio is 19%, the sieve hole is a round hole, the diameter is 1 mm, and the ratio of the diameter of the opening region to the outer diameter of the sieve plate 27 is 1.
  • Corn cobs with a particle size range of 10-20 mm were placed in an excess of aqueous medium.
  • the composition of the aqueous medium was shown in Table 2.
  • the soaked corn cob diced was removed from the aqueous solution and placed on the lower sieve.
  • the upper layer is flattened, the thickness of the material layer is 20cm, and the temperature is immersed for 12h at room temperature.
  • the culture method is the same as the culture step in the first embodiment.
  • the specific parameters are as follows: constant temperature circulating water of 36 °C is continuously introduced into the jacket, and the culture temperature is maintained at 36 ° C, air.
  • the flow rate is 0.8L/min; when the main tank body 1 is put down horizontally for the first time, the rotary drive device 49 drives the main tank body 1 to rotate, the rotation speed is 10 rpm, and the rotation is continued for 10 minutes; respectively, when the culture is carried out to 36h and 72h, respectively, the main When the can body 1 was placed in a horizontal state, the rotation speed was 1 rpm, and the rotation was continued for 3 times; a total of 96 hours was cultured. After the completion of the culture, the temperature of the constant temperature circulating water was adjusted to 40 ° C, the air flow rate was 0.8 L/min, and the mixture was dried for 24 hours, and the apparatus was placed at 25 ° C for storage.
  • the specific steps of the seeding process are the same as those in the first embodiment.
  • the specific parameters are as follows: when the main tank body 1 is placed in a horizontal state, the rotation speed is 10 rpm, and the rotation is continued for 10 minutes.
  • the culture of Saccharomyces cerevisiae. was carried out using the apparatus described in Example 1, and the culture method was the same as that of the culture and seeding steps in Example 1.
  • the components of the aqueous medium solution are shown in Table 3.
  • the material of the lever type trailer is not required to withstand high temperature steam; (2) only the main tank body is sterilized into the sterilizer during sterilization, the bracket (2) is not sterilized; (3) the main tank body 1 Rotate by manual rotation; (4) The jacket temperature of the culture process is 28 °C; (5) The flow rate of the air passing through the culture process is controlled at 0.2 L/min; (6) The culture process is carried out for 72 h, of which 12 h and 36 h. Each revolution was mixed 3 times at a speed of 1 rpm.
  • the tank shell is designed and manufactured according to the corresponding specifications of the special equipment pressure vessel, and the pressure gauge is installed; (2) the vehicle is in an upright state during sterilization, and is not placed down; (3) from the air intake filter during sterilization
  • the steam passing through the tank at 121 ° C and 0.1 MPa gauge was maintained for 20 minutes without using a sterilizer; (4) the main tank 1 was automatically rotated by means of a rotary drive device.
  • the jacket temperature of the culture process is 37 °C;
  • the flow rate of the air passing through the culture process is controlled at 0.5 L/min; (6)
  • the culture process is carried out for 72 hours, wherein the rotation is mixed for 2 times at 12 h and 36 h, and the speed is 1 rpm. .
  • the device of the present invention is relatively simple, in particular, the mixing of materials is carried out by means of a combination of a vehicle-mounted tank body and a rotary body, thereby realizing the free-slewing mixing of the tank body in two different postures of upright and horizontal; the sieve plate of the device.
  • the use of non-full-opening sieve plate can fix the position of materials and bacteria when the main tank body is in a horizontal position, and restrict the movement of materials and bacteria, so that the mixing of materials and bacteria is more uniform; the device is not only quick to use, but also convenient to move.
  • the stirring system is omitted, and the manufacturing cost of the device is greatly saved.

Abstract

一种固态生物反应装置，包括主罐体(1)，所述装置还包括支撑主罐体(1)的支架(2)，所述支架(2)使主罐体(1)在水平位置实现回转，在竖直位置实现静置培养。该装置相对简单，特别是物料的混合采用车载罐体与回转相结合的方式，实现了罐体在直立和水平两种不同姿态之间的自由转换；且该装置在直立姿态下进行装料、接种、培养和移种等工作，在水平姿态下完成灭菌和物料的混合等工作，不仅使用快捷，移动方便，而且省去了占用大量制造成本的搅拌系统，易于在大规模生产中使用。

Description

一种固态生物反应装置、使用方法及其应用 技术领域
本发明属于生物工程领域，具体涉及一种固态生物反应装置、使用方法及其应用。
背景技术
丝状菌是许多发酵产品的生产菌，在抗生素、有机酸、酶制剂和生物农药等行业广泛应用。孢子接种是丝状菌发酵常用的接种方式，在发酵生产中，丝状菌的孢子菌种一般通过表面培养(如固体培养)制备，例如柠檬酸行业普遍采用三角瓶静置扣瓶法固态培养制备黑曲霉的麸曲孢子，其简要工艺流程为：新鲜麸皮装入三角瓶灭菌，接种后上架静置培养，定时扣瓶，培养完成后干燥保存，使用时直接接种种子罐或发酵罐。
按照近年我国柠檬酸年产量一般在100～120万吨的规模估算，年使用麸曲孢子百万瓶以上。然而，规模化生产中大量使用三角瓶培养麸曲孢子，有一些难以克服的问题，例如：需要很大的恒温曲房；制备过程手工劳动量大，耗费大量人工；难以逐瓶检验孢子的质量和是否染菌，孢子的批间稳定性不易控制；麸曲保存过程易染菌；接种过程劳动量大，耗时长，染菌几率高。可见，由于目前麸曲孢子的制备过程尚未实现机械化，手工操作的制种方式，生产效率低，因此，丝状菌孢子的规模化制备技术一直是发酵行业亟待解决的共性问题。
实现丝状菌孢子规模化制备的关键在于开发符合工艺要求的设备，但国内专门用于丝状菌纯孢子规模化制备的装置尚属空白，国际上制造和销售此类装置的企业也很少。目前，市场上仅见的是奥地利奥高布殊生物商品公司(Vogelbusch Biocommodities GmbH)的孢子箱(VB Spore Box)。该设备采用多 层浅盘培养，真空收集纯孢子的技术路线，孢子的培养、干燥和收集都在封闭的系统中进行，生产能力为每批次0.8～1.4kg黑曲霉纯孢子，周期14天。然而，由于该装置使用化学熏蒸灭菌，灭菌剂必须排净以免影响微生物的生长，而排出的残余灭菌剂又需中和处理以防环境污染；琼脂培养基单独灭菌后再无菌地移入箱式培养装置，转移过程的染菌风险较高；清洗和准备的时间长，手工劳动量依然较大；琼脂培养基的成本也较高；此外，负压收集孢子的方式也会带来较高的染菌风险。
CN 104630020 A公开了一种用于规模化制备丝状菌孢子菌种的固态生物反应装置，该装置制备的干燥孢子粉，既可以立即用于发酵罐接种，又可以长时间无菌保存在孢子汇集瓶中。然而，对于一些小规模的企业和小型的发酵罐而言，为了提高孢子菌种制备过程机械化程度的同时能节省投资，还需要结构更加简单、紧凑，制造成本更低，使用更加灵活的丝状菌孢子小型制备装置。特别是装置的搅拌系统，如能简化设计，则可大幅度节约该装置的制造成本。CN 203559041 U公开了一种可移动葡萄酒发酵罐，该装置适用于葡萄酒发酵，其移动功能是水平移动，不能实现罐体姿态的旋转，也不具备回转混合的功能，因此不适于通过固态生物反应制备丝状菌孢子。CN 102924146 A公开了一种可移动转鼓式堆肥反应器，该反应器的设计用途属于固态生物反应器，装置通过转鼓的形式实现回转混合，但其移动方式限于水平移动，转动方式限于固定轴线转动，不能实现容器空间姿态的改变，也不适于通过固态生物反应制备纯培养的丝状菌孢子菌种。
考虑到上述问题，有必要开发一种新型的丝状菌孢子规模化制备装置，在实现孢子大规模机械化制备的同时解决上述问题和不足。
发明内容
本发明提供了一种固态生物反应装置、使用方法及其应用，目的是为了在小型固态反应装置中通过空间姿态的改变和容器的回转，实现物料的低强度混合，同时又能满足培养和物料混合对装置姿态的不同要求，且简化设计，节约成本。使本发明不仅能用于丝状菌孢子的规模化制备，还能用于一般微生物固态培养和固态发酵。
为达此目的，本发明采用以下技术方案：
一方面，本发明提供了一种固态生物反应装置，包括主罐体，所述装置还包括支撑主罐体的支架，所述支架能改变主罐体的姿态，以使主罐体在水平位置实现回转，在竖直位置实现静置培养；
其中，所述主罐体处于水平位置时，可手动拨动或以驱动装置自动带动主罐体转动；所述主罐体处于竖直位置时，主罐体静置培养。
本发明中，通过本发明的固态生物反应装置实现了罐体在直立和水平两种不同姿态下的自由回转混合，通过将主罐体放倒呈水平状态，可旋转罐体，可控转速，使罐内物料与接入的菌种充分混合，并可在之后的培养过程中定期回转混合以使培养物均匀生长，无需搅拌桨、轴封等部件，避免了通过搅拌浆等方式搅拌罐内物料从而破坏培养物的情况。
优选地，所述主罐体顶端中心设置有回转轴；
所述支架包括梁，用于与主罐体的回转轴相连；
所述支架设置有支座，用于支撑主罐体。
优选地，所述主罐体内部设置有至少一块筛板，用于保持物料位置
优选地，所述筛板的开孔率为1-90％，优选为1-60％。
优选地，所述筛板的外缘与主罐体的内壁密封连接，所述的密封连接可以是柔性密封也可以是刚性密封。
本发明中，所述筛板可以不是全开孔筛板，环形的非开孔区在高温灭菌过程中能在侧壁内保持部分游离的液体培养基，保证液体培养基能被培养基质颗粒充分吸收膨胀，在接种后的混合过程中保证种子液与培养基质有充分的接触并被充分吸收。
优选地，所述筛板的开孔区直径与筛板外直径的比例为0.1-1，例如可以是0.1、0.2、0.3、0.4、0.5、，055、0.6、0.7、0.8、0.9或1，优选为0.5-0.9。
优选地，所述筛板上的开孔可以但不限于圆孔和/或长圆孔槽，还可以为椭圆孔、三角形孔、多边形孔或不规则形孔。
优选地，所述圆孔的直径为0.2-20mm，例如可以是0.2mm、0.3mm、0.4mm、0.5mm、0.6mm、0.8mm、1mm、2mm、3mm、4mm、5mm、6mm、8mm、10mm、12mm、15mm、16mm、18mm、19mm或20mm，优选为0.5-3mm。
优选地，所述长圆孔槽的宽度为0.2-20mm，例如可以是0.2mm、0.3mm、0.4mm、0.5mm、0.6mm、0.8mm、1mm、2mm、3mm、4mm、5mm、6mm、8mm、10mm、12mm、15mm、16mm、18mm、19mm或20mm，优选为0.5-3mm。
本发明中，不同的菌株对通风量要求不同，筛板的开孔率可根据菌种对通风量的要求进行开孔，并调整开孔率和孔径大小。
优选地，所述主罐体自上而下包括端盖、筒体和下封头；
所述端盖包括设置于端盖顶端中心的带有销孔的回转轴，和设置于端盖上的尾气过滤器、液体过滤器与接种口；
所述筒体包括设置于筒体内壁上的视镜，和设置于筒体下部的温度传感器与罩在温度传感器外部的温度传感器套管；
所述下封头包括设置于下封头底端中部的出料阀，和设置于下封头上的进 气过滤器。
优选地，所述视镜为圆形或条形，优选为条形。
优选地，所述进气过滤器与下封头之间设置有止逆阀。
优选地，所述进气过滤器的上游设置有管接头，用于连接压缩空气管路。
优选地，所述尾气过滤器、液体过滤器和进气过滤器的孔径为0.1-0.22μm，例如可以是0.1μm、0.15μm、0.2μm、0.21μm或0.22μm，优选为0.2μm。
优选地，所述主罐体包括设置于端盖与筒体内部交界处的上筛板，和设置于筒体与下封头内部交界处的下筛板。
本发明中，所述装置可以配备包括1块上筛板和1块下筛板的1套筛板，也可以配备多套具有不同的开孔区直径，开有不同形状和不同几何尺寸的孔，并具有不同开孔率的筛板，以适用于不同的菌种和物料。
优选地，所述筒体上沿设置有上限位环，所述下封头上沿设置有下限位环。
优选地，所述筒体的外部设置有夹套，优选为筒体和下封头的外部设置有夹套，所述夹套包括设置于夹套下部的进水管和设置于夹套上部的出水管。
本发明中，夹套可覆盖筒体和下封头上全部无部件的可裸露部分，也可以仅覆盖筒体部分，也可以只覆盖下封头。
优选地，所述端盖与尾气过滤器之间设置有尾气调节阀。
优选地，所述端盖与液体过滤器之间设置有隔离阀。
优选地，所述支架能改变主罐体的姿态，可以为载车，优选为杠杆式拖车；
所述杠杆式拖车包括底座和立柱，所述底座和立柱的连接处设有车轮，所述底座和立柱整体以车轮的轴线为回转轴旋转，，所述底座和立柱随车轴旋转，所述车轴可以是一只独立车轴也可以是两只车轴；
所述底座包括竖直支撑腿和设置于底座上方的环形支座，所述环形支座用 于支撑所述主罐体；
所述立柱包括水平支撑腿，和与水平支撑腿相连的梁，用于与主罐体相连。
优选地，所述立柱内侧设置有回转驱动装置，用于驱动所述主罐体回转。
本发明中，所述回转驱动装置可以电动驱动或气动，转速可调节。
作为优选技术方案，所述水平支撑腿和竖直支撑腿也可以是带有锁止装置的脚轮。
本发明中，所述水平支撑腿和竖直支撑腿可以通过螺纹调节，在直立或水平位置时，都能保证主罐体的稳定。
本发明中，主罐体与杠杆式拖车相连，不仅可以通过主罐体可以随杠杆式拖车改变水平和竖直的位置，便于物料和菌悬液混合；同时，通过杠杆式拖车可以将主罐体轻松地移动到所需位置而移种，无需多加额外的操作步骤，降低了染菌的可能。
优选地，所述环形支座的上表面设置有滑动轴承。
优选地，所述立柱内侧设置有托轮，用于所述主罐体水平放置和水平回转的侧壁支撑。
本发明中，当装置放倒处于水平位置时，托轮用于承载主罐体的重量，同时确保主罐体能自由转动，托轮的高度可以通过托轮座上的螺栓调节，使之能与罐壁紧密贴合并能360°回转。
优选地，所述梁开有竖直的通孔，所述通孔内穿设一根固定主罐体上回转轴的套筒。
优选地，所述梁开有一个与所述通孔垂直相交的水平螺纹孔，用于安装紧固螺钉固定所述套筒。
优选地，所述套筒开有销孔1和销孔2，用于安装固定销。
优选地，所述套筒下端的内表面设置有滑动轴承。
优选地，所述主罐体和支架的制作材料为能耐受121℃、0.1MPa蒸汽灭菌的材料，可以选自但不限于不锈钢、碳钢、有色金属、轻质合金、塑料、陶瓷、搪玻璃或玻璃中的任意一种或至少两种的混合。
优选地，所述端盖为椭圆形、球冠形、球形、碟形或平板，优选为椭圆形或碟形，进一步优选为椭圆形。
优选地，所述下封头为椭圆形、球冠形、球形、碟形或锥形，优选为锥形。
第二方面，本发明提供了一种固态培养微生物的方法，采用如第一方面所述的固体生物反应装置，所述方法包括以下步骤：
(1)将浸泡液体培养基营养成分的颗粒状培养基质放入主罐体中，将主罐体进行灭菌；
(2)灭菌后进行接种，将主罐体放倒呈水平状态，主罐体旋转混合，再将主罐体直立起恒温培养；
任选地，培养期间进行多次混合；
(3)培养完成后，可以通风至培养物完全干燥，封存备用；
本发明，若培养发酵罐用的菌种，则将培养完成后得到培养物移种到发酵罐中进行扩大培养。
作为优选技术方案，向发酵罐移种的具体步骤包括：打开隔离阀和尾气调节阀，通过过滤除菌的方式向罐内加水或水溶液，之后关闭隔离阀和尾气调节阀，将主罐体放倒，持续进行回转混合的操作，直至通过视镜可见大部分菌已被洗下。将主罐体立起，拧下紧固螺钉，在梁的螺纹孔中插入销子，使整个主罐体悬挂在较高位置。再通过出料阀与发酵罐顶接种阀之间连接移种管，通过阀门向其通入温度不低于121℃，压力不低于0.1MPa(表压)的蒸汽，维持至 少20min以对该段管路进行灭菌。在管接头处连接压缩空气管，通入压力高于发酵罐罐压的压缩空气，将装置内的菌种悬液压入发酵罐中；或移种方式还可以按照以下步骤进行：在安装完移种连接管并灭菌后，关闭尾气调节阀和隔离阀，打开主罐体底阀和发酵罐顶移种阀，使主罐体与发酵罐上部空气连通，之后降低发酵罐的罐压，利用压差将菌种悬液移入发酵罐。可以反复加水若干次，将菌种尽可能彻底地移入发酵罐。移种完毕，关闭发酵罐顶移种阀，拆除移种管道。
优选地，步骤(1)所述方法具体包括以下步骤：
(1a)打开端盖，将用液体培养基营养成分浸泡过的颗粒状培养基质放入主罐体，盖上端盖；
(1b)将主罐体与支架组装固定：所述主罐体的下限位环与所述杠杆式拖车的环形支座上的滑动轴承紧密贴合，再将套筒穿过梁上的竖直通孔下放，套住主罐体的回转轴，对准销孔2后插入销，之后将紧固螺钉通过螺纹孔旋入梁并拧紧，顶住套筒，将套筒和梁紧固为一体；
(1c)主罐体随杠杆式拖车放倒呈水平状态，打开尾气调节阀和隔离阀，进行灭菌。
优选地，步骤(2)所述方法具体包括以下步骤：
(2a)灭菌后，在管接头处插上压缩空气管，通入压缩空气吹干进气过滤器、尾气过滤器和液体过滤器，同时培养基质冷却，关闭隔离阀，微开尾气调节阀，在火焰保护下，自接种口接入待培养微生物的液体种子；
(2b)接种后，将主罐体随杠杆式拖车放倒呈水平状态，拔出销孔的销子，用手拨动主罐体使之旋转，从而使罐内物料与接入的液体种子悬液充分混合；
(2c)混合后重新插入销子，将主罐体立起，通入压缩空气进行培养，通 过进水管和出水管向夹套中通入与培养温度相同的恒温水，以维持培养物的温度；
(2d)培养过程中间歇性地将主罐体放倒，重复回转混合的操作数次。
(2e)培养结束后，可以从空气管接头通入压缩空气，吹干培养物。
第三方面，本发明提供了一种如第一方面所述的固体生物反应装置，采用如第二面所述的方法在培养微生物或固态发酵中的应用。
本发明反应装置不仅能用于丝状菌孢子的规模化制备，还能用于一般的微生物固态培养和固态发酵。
本发明中，所述微生物可以选自但不限于丝状菌、酵母菌或细菌。
与现有技术相比，本发明具有如下有益效果：
(1)本发明装置相对简单，特别是物料的混合采用车载罐体与回转相结合的方式，实现了罐体在直立和水平两种不同姿态下的自由回转混合；
(2)本发明装置的筛板采用非全开孔筛板，能在主罐体呈水平位置时，固定物料和菌的位置，限制物料和菌的移动，使得物料和菌的混合更均匀；
(3)本发明装置可用于便捷地固体培养微生物，尤其是制备丝状菌孢子和各种单细胞微生物，并且制备的孢子既能以悬浮液的形式方便且无菌地转移到发酵罐中，又能与培养基质一同干燥后长时间地保存在该装置中备用，孢子的制备和转接过程比麸曲大为简化；制备的单细胞微生物也能以菌悬液的形式方便且无菌地转移到发酵罐中。
(4)本发明装置在直立姿态下进行装料、接种、培养、加水和移种工作，在水平姿态下完成灭菌和物料的回转混合工作，不仅使用快捷，移动方便，而且省去了占用大量制造成本的搅拌系统，维护费用低。
附图说明
图1为本发明的主罐体示意图；
图2为本发明载车主视图；
图3为本发明载车左视图；
图4为本发明载车俯视图；
图5为本发明接种/培养状态的示意图；
图6为本发明灭菌/回转混合状态的示意图；
图7为本发明移种状态的示意图；
图8为本发明的四阀移种装置示意图；
图9为本发明筛板及其开孔区示意图；
其中，图中1-主罐体；3-端盖；4-罐体；5-下封头；6-尾气过滤器；7-液体过滤器；8-接种口；9-视镜；10-温度传感器；11-温度传感器套管；12-夹套；13-循环水的进水管；14-循环水的出水管；15-进气过滤器；16-出料阀；17-止逆阀；18-回转轴；19-销孔；20-上限拉环；21-下限拉环；22-上筛板；23-下筛板；24-管接头；25-尾气调节阀；26-隔离阀；27-筛板；28-车轴；29-底座；30-环形支座；31-立柱；32-梁；33-水平支撑腿；34-竖直支撑腿；35-竖直通孔；36-套筒；37-销孔1；38-销孔2；39-车轮；40-托轮；41-螺纹孔；42-滑动轴承1；43-滑动轴承2；44-接种阀；45-移种管；46-阀门；47-阀门；48-发酵罐；49-驱动装置。
具体实施方式
为更进一步阐述本发明所采取的技术手段及其效果，以下结合附图并通过具体实施方式来进一步说明本发明的技术方案，但本发明并非局限在实施例范围内。
实施例1：一种固态生物反应装置
如图1所示，是本发明实施例所述固态生物反应装置的示意图，所述装置 包括主罐体1和杠杆式拖车2，所述杠杆式拖车2使主罐体1在水平位置回转，在竖直位置静置培养；
其中，所述主罐体1处于水平位置时，可手动拨动或驱动装置49自动带动主罐体1转动；所述主罐体1处于竖直位置时，主罐体1静置培养，如图6所示。
所述主罐体1自上而下包括端盖3、筒体4和下封头5；
所述端盖3包括设置于端盖3顶端中心的带有销孔19的回转轴18，和设置于端盖3上的尾气过滤器6、液体过滤器7与接种口8；所述端盖3与尾气过滤器6之间设置有尾气调节阀25；所述端盖3与液体过滤器7之间设置有隔离阀26。
所述筒体4包括设置于筒体4内部的视镜9，和设置于筒体4下部的温度传感器10与罩在温度传感器10外部的温度传感器套管11；所述筒体4上沿设置有上限位环20。
所述下封头5包括设置于下封头5底端中心的出料阀16，和设置于下封头5上的进气过滤器15；所述进气过滤器15与下封头5之间设置有止逆阀17；所述进气过滤器15的上游设置有管接头24，用于连接压缩空气管路；，所述下封头5上沿设置有下限位环21。
所述主罐体1内部设置端盖3与筒体4内部交界处的上筛板22，和设置于筒体4与下封头5内部交界处的下筛板23，用于保持物料位置；所述筛板27的开孔率为1-90％。
所述杠杆式拖车包括底座29和立柱31，所述底座29和立柱31的连接处穿设有一车轴28，所述底座29和立柱31随车轴28旋转，所述车轴28两端设有车轮39；
所述底座29包括竖直支撑腿34和设置于底座29上方的环形支座30，所述环形支座30用于支撑所述主罐体1；所述环形支座30的上表面设置有滑动轴承42。
所述立柱31包括水平支撑腿33，和与水平支撑腿33相连的梁32，用于与主罐体4相连，和设置于水平支撑腿33右侧的托轮40，用于所述主罐体1水平放置和水平回转的侧壁支撑。
所述梁32开有竖直的通孔35，所述通孔35内穿设一根固定主罐体1上回转轴18的套筒36；所述梁32开有一个与所述通孔35垂直相交的水平螺纹孔41，用于安装紧固螺钉固定所述套筒36；所述套筒36开有销孔1(37)和销孔2(38)，用于安装固定销；所述套筒36下端的内表面设置有滑动轴承43。
所述装置的使用方法，包括以下步骤：
首先打开端盖3，将用营养成分的液体培养基充分浸泡培养基质颗粒放入主罐体1，盖上端盖3；将主罐体1与支架2组装固定：所述主罐体1的下限位环21与所述杠杆式拖车2的环形支座30上的滑动轴承42紧密贴合，再将套筒36穿过梁32上的竖直通孔35下放，套住主罐体1的回转轴18，对准销孔38后插入销，之后将紧固螺钉通过螺纹孔41旋入梁32并拧紧，顶住套筒36，将套筒36和梁32紧固为一体；主罐体1随杠杆式拖车2放倒呈水平状态，打开尾气调节阀25和隔离阀26，进行灭菌。
其次，灭菌后，在管接头24处插上压缩空气管，通入压缩空气吹干进气过滤器15、尾气过滤器6和液体过滤器7，同时培养基质冷却，关闭隔离阀26，微开尾气调节阀25，在火焰保护下，自接种口8接入待培养微生物的液体种子；接种后，将主罐体1随杠杆式拖车2放倒呈水平状态，拔出销孔38的销子，用 手拨动主罐体1使之旋转，从而使罐内物料与接入的液体种子充分混合；混合后重新插入销子，将主罐体1立起，通入压缩空气进行培养，通过进水管13和出水管14向夹套12中通入与培养温度相同的恒温水，以维持培养物的温度；培养过程中间歇性地将主罐体1放倒，重复回转混合的操作数次；
最后，将培养完成后得到培养物移种到发酵罐48中进行扩大培养，移种的具体步骤包括：打开隔离阀25和尾气调节阀26，通过过滤除菌的方式向罐内加水或水溶液，之后再关闭尾气调节阀25和隔离阀26，将主罐体1放倒，持续进行回转混合的操作，直至通过视镜9可见大部分菌已被洗下。将主罐体1立起，拧下紧固螺钉，在梁的螺纹孔中插入销子，使整个主罐体1悬挂在较高位置。再通过出料阀16与发酵罐顶接种阀44之间连接移种管45，通过阀门46、47向其中通入温度不低于121℃，压力不低于0.1MPa(表压)的蒸汽，维持至少20min以对该段管路进行灭菌。在管接头24处连接压缩空气管，通入压力高于发酵罐48罐压的压缩空气，将装置内的菌悬液压入发酵罐48中。可以反复加水若干次，将菌尽可能彻底地移入发酵罐48。移种完毕，关闭发酵罐顶阀门44，拆除移种管道。或移种方式还可以按照以下步骤进行：在安装完移种管45并灭菌后，关闭尾气调节阀25和隔离阀26，打开主罐体1下部的出料阀16和发酵罐顶阀门44，使主罐体1与发酵罐48上部空气连通，之后降低发酵罐48的罐压，利用压差将菌悬液移入发酵罐48。
在具体实施例1中，筛板27还可以设置多层分布在主罐体1中，所述筛板27的开孔区直径与筛板27外直径的比例为0.1-1，优选为0.5-0.9；所述筛板27上的开孔为圆孔和/或长圆孔槽；所述圆孔的直径为0.5-20mm，优选为1-10mm；所述长圆孔槽的宽度为0.5-20mm，优选为1-10mm。
在具体实施例1中，所述视镜9为圆形或条形，优选为条形。
在具体实施例1中，所述尾气过滤器6、液体过滤器7和进气过滤器15的孔径为0.2-0.22μm，优选为0.2μm。
在具体实施例1中，所述筒体4的外部设置有夹套12，优选为筒体4和下封头5的外部设置有夹套12，所述夹套12包括设置于夹套12下部的进水管13和设置于夹套12上部的出水管14。
在具体实施例1中，所述水平支撑腿33和竖直支撑腿34也可以是带有高度调整装置和锁止装置的脚轮。
在具体实施例1中，所述主罐体1和载车2的制作材料为耐受121℃、0.1MPa蒸汽的材料，可以选自但不限于不锈钢、碳钢、有色金属、轻质合金、塑料、陶瓷、搪玻璃或玻璃中的任意一种或至少两种的混合。
在具体实施例1中，所述端盖3为椭圆形、球冠形、球形、碟形或平板，优选为椭圆形或碟形，进一步优选为椭圆形。
在具体实施例1中，所述下封头5为椭圆形、球冠形、球形、碟形或锥形，优选为锥形。
实施例2
对黑曲霉(Aspergillus niger)的孢子培养采用实施例1所述的装置，培养方法同实施例1中的培养步骤，培养基水溶液的成分见表1，其中，培养过程中步骤如下：
主罐体1全容积30L，材质为316L不锈钢，端盖3为椭圆形，下封头5为锥顶角120°的锥形，夹套覆盖筒体4和下封头5全部裸露无附件部分；
筛板27还可以设置多层分布在主罐体1中，所述筛板27的开孔率为29％，开孔区直径与筛板27外直径的比例为0.86；所述筛板27上的开孔为圆孔，所 述圆孔的直径为3mm。
区粒径范围为5-10mm的玉米芯颗粒置于过量的培养基水溶液中，培养基水溶液的成分见表1，将浸透的玉米芯颗粒从培养基水溶液中捞出，放在下筛板上摊平，料层厚度10cm，室温浸泡12h，培养方法同实施例1中的培养步骤，具体参数如下：向夹套内持续通入35℃的恒温循环水，保持培养温度为35℃，空气流量0.5L/min；首次将主罐体1放倒呈水平状态时，手动旋转主罐体1，旋转速度10rpm，持续转动10min；分别在培养到48h和96h时，再将主罐体1放倒呈水平状态时，旋转速度1rpm，持续转动2圈；一共培养120h。培养结束后，将恒温循环水温度调高到38℃，空气流量0.5L/min，干燥24h，将装置放置在25℃保存。
移种过程具体步骤同实施例1中的培养步骤，具体参数如下：主罐体1放倒呈水平状态时，旋转速度10rpm，持续转动10min。
表1
成分 浓度(g/L)
葡萄糖 50
NaNO3 2
MgSO4·7H2O 0.5
K2HPO4 1
KCl 0.5
FeSO4·4H2O 0.01
实施例3
对曲霉属微生物(Aspergillus sp.)的孢子培养采用实施例1所述的装置，培养方法同实施例1中的培养步骤，培养基水溶液见表2，其中，培养过程中步骤如下：
主罐体1全容积45L，材质为304不锈钢，端盖3为碟形，下封头5为球形，夹套覆盖筒体4和下封头5全部裸露无附件部分；
筛板27还可以设置多层分布在主罐体1中，所述上筛板22开孔率为39％，筛孔为圆孔，直径2mm，开孔区直径与筛板27外直径的比例为0.79，下筛板开孔率为19％，筛孔为圆孔，直径1mm，开孔区直径与筛板27外直径的比例为1。
取粒径范围为10-20mm的玉米芯切块置于过量的培养基水溶液中，培养基水溶液的成分见表2，将浸透的玉米芯切块从培养基水溶液中捞出，放在下筛板上摊平，料层厚度20cm，室温浸泡12h，培养方法同实施例1中的培养步骤，具体参数如下：向夹套内持续通入36℃的恒温循环水，保持培养温度为36℃，空气流量0.8L/min；首次将主罐体1放倒呈水平状态时，回转驱动装置49带动主罐体1旋转，旋转速度10rpm，持续转动10min；分别在培养到36h和72h时，再将主罐体1放倒呈水平状态时，旋转速度1rpm，持续转动3圈；一共培养96h。培养结束后，将恒温循环水温度调高到40℃，空气流量0.8L/min，干燥24h，将装置放置在25℃保存。
移种过程具体步骤同实施例1中的培养步骤，具体参数如下：主罐体1放倒呈水平状态时，旋转速度10rpm，持续转动10min。
表2
成分 浓度(g/L)
葡萄糖 40
NaNO3 1.6
MgSO4·7H2O 0.4
K2HPO4 0.8
KCl 0.4
FeSO4·4H2O 0.008
实施例4
培养酿酒酵母(Saccharomyces cerevisiae.)采用实施例1所述的装置，培养方法同实施例1中的培养和移种步骤，培养基水溶液的成分见表3。
其中，(1)杠杆式拖车的材料不要求耐受高温蒸汽；(2)灭菌时只将主罐体搬入灭菌器内灭菌，支架(2)不灭菌；(3)主罐体1采取手动旋转的方式旋转；(4)培养过程夹套温度28℃；(5)培养过程通入空气的流量控制在0.2L/min；(6)培养过程共进行72h，其中在12h和36h各回转混合3圈，速度1rpm。
表3
成分 浓度(g/L)
葡萄糖 20
酵母浸出物 10
蛋白胨 20
实施例5
培养枯草芽孢杆菌(Bacillus subtilis.)采用实施例1所述的装置，培养方法 同实施例1中的培养和移种步骤，培养基水溶液的成分见表4。
其中，(1)罐体外壳按照特种设备压力容器的相应规范设计制造，加装压力表；(2)灭菌时车处于直立状态，不放倒；(3)灭菌时从进气过滤器向罐内通121℃，0.1MPa表压的蒸汽，维持20min，不使用灭菌器；(4)主罐体1采用回转驱动装置带动的方式自动旋转。(4)培养过程夹套温度37℃；(5)培养过程通入空气的流量控制在0.5L/min；(6)培养过程共进行72h，其中在12h和36h各回转混合2圈，速度1rpm。
表4
成分 浓度(g/L)
NaCl 10
酵母浸出物 5
蛋白胨 10
综上所述，本发明装置相对简单，特别是物料的混合采用车载罐体与回转相结合的方式，实现了罐体在直立和水平两种不同姿态下的自由回转混合；该装置的筛板采用非全开孔筛板，能在主罐体呈水平位置时，固定物料和菌的位置，限制物料和菌的移动，使得物料和菌的混合更均匀；该装置不仅使用快捷，移动方便，且省去了搅拌系统，大幅节省该装置的制造成本。
申请人声明，本发明通过上述实施例来说明本发明的详细方法，但本发明并不局限于上述详细方法，即不意味着本发明必须依赖上述详细方法才能实施。所属技术领域的技术人员应该明了，对本发明的任何改进，对本发明产品各原料的等效替换及辅助成分的添加、具体方式的选择等，均落在本发明的保护范围和公开范围之内。

Claims (19)

  1. 一种固态生物反应装置，包括主罐体(1)，其特征在于，所述装置还包括支撑主罐体(1)的支架(2)，所述支架(2)使主罐体(1)在水平位置实现回转，在竖直位置实现静置培养。
  2. 根据权利要求1所述的装置，其特征在于，所述主罐体(1)顶端中心设置有回转轴(18)；
    所述支架(2)包括梁(32)，用于与主罐体(1)的回转轴(18)相连；
    所述支架(2)设置有支座(30)，用于支撑主罐体(1)。
  3. 根据权利要求1或2所述的装置，其特征在于，所述主罐体内部设置有至少一块筛板(27)，用于保持物料位置。
  4. 根据权利要求3所述的装置，其特征在于，所述筛板的开孔率为1-90％。
  5. 根据权利要求3所述的装置，其特征在于，所述筛板的开孔率为1-60％。
  6. 根据权利要求3所述的装置，其特征在于，所述筛板的开孔区直径与筛板外直径的比例为0.1-1。
  7. 根据权利要求3所述的装置，其特征在于，所述筛板的开孔区直径与筛板外直径的比例为0.5-0.9。
  8. 根据权利要求3所述的装置，其特征在于，所述筛板上的开孔为圆孔和/或长圆孔槽。
  9. 根据权利要求8所述的装置，其特征在于，所述圆孔的直径为0.2-20mm。
  10. 根据权利要求8所述的装置，其特征在于，所述圆孔的直径为0.5-3mm。
  11. 根据权利要求8所述的装置，其特征在于，所述长圆孔槽的宽度为0.2-20mm。
  12. 根据权利要求8所述的装置，其特征在于，所述长圆孔槽的宽度为0.5-3mm。
  13. 根据权利要求1-12中任一项所述的装置，其特征在于，所述主罐体自上而下包括端盖(3)、筒体(4)和下封头(5)；
    所述端盖(3)包括设置于端盖(3)顶端中心的带有销孔(19)的回转轴(18)，和设置于端盖(3)上的尾气过滤器(6)、液体过滤器(7)与接种口(8)；
    所述筒体(4)包括设置于筒体(4)内壁上的视镜(9)，和设置于筒体(4)下部的温度传感器(10)与罩在温度传感器(10)外部的温度传感器套管(11)；
    所述下封头(5)包括设置于下封头(5)底端中部的出料阀(16)，和设置于下封头(5)上的进气过滤器(15)；
    优选地，所述进气过滤器(15)与下封头(5)之间设置有止逆阀(17)；
    优选地，所述进气过滤器(15)的上游设置有管接头(24)，用于连接压缩空气管路；
    优选地，所述尾气过滤器(6)、液体过滤器(7)和进气过滤器(15)的孔径为0.1-0.22μm，优选为0.2μm。
  14. 根据权利要求1-13中任一项所述的装置，其特征在于，所述主罐体(1)包括设置于端盖(3)与筒体(4)内部交界处的上筛板(22)，和设置于筒体(4)与下封头(5)内部交界处的下筛板(23)；
    优选地，所述筒体(4)上沿设置有上限位环(20)，所述下封头(5)上沿设置有下限位环(21)；
    优选地，所述筒体(4)的外部设置有夹套(12)，优选为所述筒体(4)和下封头(5)的外部设置有夹套(12)，所述夹套(12)包括设置于夹套(12)下部的进水管(13)和设置于夹套(12)上部的出水管(14)；
    优选地，所述端盖(3)与尾气过滤器(6)之间设置有尾气调节阀(25)；
    优选地，所述端盖(3)与液体过滤器(7)之间设置有隔离阀(26)。
  15. 根据权利要求1-14中任一项所述的装置，其特征在于，所述支架(2)为载车，优选为杠杆式拖车；
    所述杠杆式拖车包括底座(29)和立柱(31)，所述底座(29)和立柱(31)的连接处设有车轮(39)，所述底座(29)和立柱(31)整体以车轮(39)的轴线为回转轴旋转；
    所述底座(29)包括竖直支撑腿(34)和设置于底座(29)上方的环形支座(30)，所述环形支座(30)用于支撑所述主罐体(1)；
    所述立柱(31)包括水平支撑腿(33)，和与水平支撑腿(33)相连的梁(32)，用于与主罐体(1)相连；
    优选地，所述环形支座(30)的上表面设置有滑动轴承(42)；
    优选地，所述立柱(31)内侧设置有托轮(40)，用于所述主罐体(1)水平放置和水平回转的侧壁支撑；
    优选地，所述立柱(31)内侧设置有回转驱动装置(49)，用于驱动所述主罐体(1)回转；
    优选地，所述梁(32)开有竖直的通孔(35)，所述通孔(35)内穿设一根固定主罐体(1)上回转轴(18)的套筒(36)；
    优选地，所述梁(32)开有一个与所述通孔(35)垂直相交的水平螺纹孔(41)，用于安装紧固螺钉固定所述套筒(36)；
    优选地，所述套筒(36)开有销孔1(37)和销孔2(38)，用于安装固定销；
    优选地，所述套筒(36)下端的内表面设置有滑动轴承(43)。
  16. 根据权利要求1-15中任一项所述的装置，其特征在于，所述主罐体(1)和支架(2)的制作材料为不锈钢、碳钢、有色金属、轻质合金、塑料、搪玻璃 或玻璃中的任意一种或至少两种的混合；
    优选地，所述端盖(3)为椭圆形、球冠形、球形、碟形或平板，优选为椭圆形或碟形，进一步优选为椭圆形；
    优选地，所述下封头(5)为椭圆形、球冠形、球形、碟形或锥形，优选为锥形。
  17. 一种固态培养微生物的方法，其特征在于，采用权利要求1-7中任一项所述的固体生物反应装置，所述方法包括以下步骤：
    (1)将浸泡液体培养基营养成分的颗粒状培养基质放入主罐体(1)中，将主罐体(1)进行灭菌；
    (2)灭菌后进行接种，将主罐体(1)放倒呈水平状态，转动主罐体(1)旋转混合，再将主罐体(1)直立起恒温培养；
    任选地，培养期间进行多次混合；
    (3)培养完成后，通风至培养物完全干燥，封存备用；
    任选地，将培养完成后得到培养物移种到发酵罐(48)中进行扩大培养。
  18. 根据权利要求17所述的方法，其特征在于，步骤(1)所述方法具体包括以下步骤：
    (1a)打开端盖(3)，将用液体培养基营养成分充分浸泡的颗粒状培养基质放入主罐体(1)，盖上端盖(3)；
    (1b)将主罐体(1)与支架(2)组装固定：所述主罐体(1)的下限位环(21)与所述杠杆式拖车(2)的环形支座(30)上的滑动轴承(42)紧密贴合，再将套筒(36)穿过梁(32)上的竖直通孔(35)下放，套住主罐体(1)的回转轴(18)，对准销孔2(38)后插入销，之后将紧固螺钉通过螺纹孔(41)旋入梁(32)并拧紧，顶住套筒(36)，将套筒(36)和梁(32)紧固为一体；
    (1c)主罐体(1)随杠杆式拖车(2)放倒呈水平状态，打开尾气调节阀(25)和隔离阀(26)，进行灭菌；
    优选地，步骤(2)所述方法具体包括以下步骤：
    (2a)灭菌后，在管接头(24)处插上压缩空气管，通入压缩空气吹干进气过滤器(15)、尾气过滤器(6)和液体过滤器(7)，同时培养基质冷却，关闭隔离阀(26)，微开尾气调节阀(25)，在火焰保护下，自接种口(8)接入待培养微生物的液体种子；
    (2b)接种后，将主罐体(1)随杠杆式拖车(2)放倒呈水平状态，拔出销孔2(38)的销子，主罐体(1)旋转，从而使罐内物料与接入的液体种子充分混合；
    (2c)混合后重新插入销子，将主罐体(1)立起，通入压缩空气进行培养，通过进水管(13)和出水管(14)向夹套(12)中通入与培养温度相同的恒温水，以维持培养物的温度；
    (2d)培养过程中间歇性地将主罐体(1)放倒，重复回转混合的操作数次；
    任选地，培养结束后，从空气管接头通入压缩空气，吹干培养物。
  19. 一种如权利要求1-15中任一项所述的固体生物反应装置，采用如权利要求16-18中任一项所述的方法在培养微生物或固态发酵中的应用。
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