CN113088431A

CN113088431A - High-activity and high-stability shiitake mushroom liquid strain fermentation system and fermentation method

Info

Publication number: CN113088431A
Application number: CN202110464883.1A
Authority: CN
Inventors: 万晓芹; 刘运航; 罗红梅; 王旭; 邓国云; 刘敏; 韩亚琴; 韦云勤
Original assignee: Guizhou Tonghui Edible Fungus Development Co ltd
Current assignee: Guizhou Tonghui Edible Fungus Development Co ltd
Priority date: 2021-04-28
Filing date: 2021-04-28
Publication date: 2021-07-09

Abstract

The invention discloses a high-activity and high-stability lentinus edodes liquid strain fermentation system and a fermentation method, wherein the fermentation method comprises the following steps: the surge tank, the one-level jar, the second grade jar, the liquid outlet and the one-level jar of surge tank are connected, the liquid outlet and the second grade jar of one-level jar are connected, the surge tank includes omnipotent spring holder, the top of omnipotent spring holder is provided with balanced splint, the top of balanced splint is fixed with a plurality of test tubes, the test tube can slide on balanced splint, and it is empty to have at least one test tube to stay, the top of all test tubes is all through the intercommunication of intercommunication frame, the intercommunication of intercommunication frame and one-level jar, can let the test tube slide on balanced splint, the weight at free adjustment balanced splint both ends, make balanced splint can remain stable, the symmetry, can be more stable when shaking the bottle, and stable shake the bottle effect and can make the better various nutrition in the absorption culture medium of mother species, the fungus that ferments out more accords with the standard, the activity of mushroom liquid fungus.

Description

High-activity and high-stability shiitake mushroom liquid strain fermentation system and fermentation method

Technical Field

The invention relates to liquid strain fermentation, in particular to a high-activity and high-stability shiitake mushroom liquid strain fermentation system and a fermentation method.

Background

The "liquid spawn" is a liquid form of edible fungus spawn produced by a submerged culture (liquid fermentation) technique in a biological fermentation tank using a liquid medium. The liquid refers to the physical state of a culture medium, and the liquid submerged culture is a fermentation engineering technology.

The traditional mushroom production generally adopts solid strains for inoculation culture, the strains mainly comprise sawdust strains, branch strains and other conventional solid strains, the production period of the method is long, the method generally needs 55-60 days, the process is complex, and the production method has low efficiency; the production process of the strain needs a large space, needs a large amount of manpower and material resources, and has high cost; the culture period of the hyphae is long, the difference of the hyphae age in the culture medium is large, when the front end hyphae are in a budding state, the hyphae on the surface of the substrate are nearly aged, the fruiting of the cultivated species is irregular, the scale production of the mushrooms cannot be achieved, and the production requirements of people cannot be met.

In the prior art, in the process of shaking the liquid spawn, staff often adjust because the counter weight at both ends of the oscillation box, and there are volume, specification difference of solution, it is difficult to make the easy weight at both ends equal, the oscillation box communicates with each other with the outside always when adjusting, has brought great puzzlement to the aseptic nature experiment of bacterial, makes the mushroom liquid spawn activity that obtains lower, comparatively unstable.

Disclosure of Invention

The invention provides a high-activity and high-stability lentinus edodes liquid strain fermentation system and a fermentation method, which can effectively solve the problems.

The invention is realized by the following steps:

a high activity, high stability mushroom liquid bacterial fermentation system includes: the liquid outlet of the oscillation box is connected with the first-stage tank, the liquid outlet of the first-stage tank is connected with the second-stage tank,

the vibration box includes omnipotent spring holder, the top of omnipotent spring holder is provided with balanced splint, the top of balanced splint is fixed with a plurality of test tubes, the test tube can slide on the balanced splint, and has at least one leave empty in the test tube, all the top of test tube all communicates through the intercommunication frame, the intercommunication frame with the inlet intercommunication of one-level jar.

As further improved, balanced splint divide into two counterpoint boards of bilateral symmetry, all be equipped with weighing transducer in the counterpoint board, a plurality of test tubes are held to the clamp in the counterpoint board, the slide is clamped to the bottom dress of test tube, the slide can drive the test tube is in slide in the counterpoint board, make both ends counterpoint board weight is kept level.

As a further improvement, the aligning plate can be adjusted in width, and after the weight is leveled, the test tube can be clamped to follow the universal spring seat to shake the bottle.

Further include the fungus crowd and detect the structure, the fungus crowd detects the structure and encircles the surface that sets up at all test tubes, the fungus crowd detects the structure and is being most peripheral all be equipped with the camera assembly on the test tube relative position, be provided with main camera and vice camera on the camera assembly, main camera is just right the test tube, vice camera is located the both sides of main camera, vice camera position adjustable and with the test tube sets up relatively.

As a further improvement, the test tube at the inner periphery is at least opposite to more than two auxiliary cameras.

As a further improvement, the second-stage tank is provided with a stirring rod, the bottom of the second-stage tank is also provided with a conveying cavity, and the conveying cavity is provided with a plurality of permeation tubes.

As further improvement, be connected with a plurality of stirring branch poles on the puddler, be provided with a plurality of rings on the infiltration pipe, be provided with a plurality of shrinkage pools between the ring, it is right after the ring is exerted pressure, the shrinkage pool can the blowout gas extremely in the second grade jar, the stirring divide the pole with the ring sets up relatively.

As a further improvement, the conveying cavity comprises an upper guide layer, the bottom of the upper guide layer is connected with a lower guide layer, and a sterile joint for connecting an external air source is arranged on the lower guide layer.

A high-activity and high-stability shiitake mushroom liquid strain fermentation method comprises the following steps:

s1; placing the culture medium into the test tube, placing the cut mushroom slant strains into the test tube, sealing the test tube, placing the container into the oscillation box,

s2; after the oscillating box is closed, the weight of the alignment plates in the oscillating box is matched by control equipment to ensure that the weights of the alignment plates at two ends are consistent,

s3; adjusting the temperature of the oscillation box, standing for 40-50h at 25 +/-2 ℃, adjusting the temperature to 17-21 ℃, starting the oscillation box, and starting oscillation culture for 60-72h to obtain shake flask strains;

s4; the shake flask strains in the test tube are all extracted through the communicating frame, are not contacted with the outside in the extracting process, are directly extracted into the primary tank, are sterilized for 2 to 3 hours under the conditions that the steam pressure is 0.15 to 0.19MPa and the temperature is 120 ℃ and 122 ℃ to obtain primary fermentation strains,

s5; introducing the primary fermentation strain into the secondary tank, stirring by a stirring rod, continuously supplying oxygen to the primary fermentation strain by a permeation tube 32 in the stirring process, introducing oxygen at 150 +/-10 m3/h, fermenting at 20 +/-2 ℃ and under the tank pressure of 0.15 +/-0.02 MPa, and culturing under the condition of 90-100 ℃ to obtain the liquid strain of the lentinus edodes.

As a further improvement, in step S2, the alignment plates at the left and right ends each leave empty one test tube, and the communication rack can fill water into the test tubes to balance the weight.

The invention has the beneficial effects that:

according to the invention, through the matching of the balance clamping plate and the test tube, the test tube can slide on the balance clamping plate, the weight of two ends of the balance clamping plate can be freely adjusted, and through the way of leaving at least one empty test tube, water can be added or pumped to the empty test tube, and the balance clamping plate is weighted, so that the balance clamping plate can be kept stable and symmetrical, and can be more stable during shaking, and the stable shaking effect can enable the mother strain to better absorb various nutrients in a culture medium, the fermented strain can better meet the standard, and the activity of the mushroom liquid strain is higher and more stable.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are required to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.

FIG. 1 is a schematic structural diagram of a high-activity and high-stability lentinus edodes liquid strain fermentation system.

Fig. 2 is a schematic structural diagram of an oscillation box of the present invention.

Fig. 3 is a schematic structural view of a balance splint of the present invention.

FIG. 4 is a schematic top view of a bacterial colony detection structure according to the present invention.

FIG. 5 is a schematic diagram of a photographing circuit of a flora detection structure according to the present invention.

Fig. 6 is a schematic top view of a secondary tank of the present invention.

Fig. 7 is a schematic structural view of a delivery lumen of the present invention.

Detailed Description

The embodiments of the present invention are within the scope of the present invention. Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.

In the description of the present invention, the terms "first" and "second" are used for descriptive purposes only and are not to be construed as indicating the purposes, technical solutions and advantages of the embodiments of the present invention, which will be described in detail and fully with reference to the accompanying drawings in the embodiments of the present invention. All other implementations that can be derived by one of ordinary skill in the art based on the embodiments of the present invention show or imply relative importance or implicitly indicate the number of technical features indicated, without inventive step. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless specifically defined otherwise.

Referring to fig. 1-7, a high-activity and high-stability shiitake mushroom liquid strain fermentation system and a fermentation method thereof include: the method comprises the following steps: an oscillating box 1, a primary tank 2 and a secondary tank 3, wherein a liquid outlet of the oscillating box 1 is connected with the primary tank 2, the liquid outlet of the first-stage tank 2 is connected with the second-stage tank 3, the oscillation box 1 comprises a universal spring seat 11, a balance clamping plate 12 is arranged above the universal spring seat 11, a plurality of test tubes 15 are fixed above the balance clamping plate 12, the test tubes 15 can slide on the balance clamping plate 12, at least one test tube 15 is left empty, the upper parts of all the test tubes 15 are communicated through a communicating rack 15, the communicating frame 15 is communicated with the liquid inlet of the primary tank 2, and after mother seeds are put into the oscillating box 1, the oscillating box 1 is closed without being opened any more, so as to avoid the infection of external bacteria until the mother seeds are filled next time, and after the strains after shaking the bottles are sterilized by a first-stage tank 2, oxygen is introduced into the bottles by a second-stage tank 3, and the bottles are stirred to obtain the mushroom liquid strains.

Wherein, the pipette position that the intercommunication frame 15 corresponds every test tube 15 is all adjustable, can move about on the intercommunication frame 15 along with test tube 15's activity, and the intercommunication frame 15 of being convenient for transfers the position, and, the bottom of pipette is big opening, can guarantee not excessively to destroy the structure of bacterial when absorbing the fungus crowd, keeps the stability of bacterial.

Specifically, the balance clamp 12 is divided into two alignment plates 121 which are bilaterally symmetrical, the weight sensors 122 are respectively arranged in the alignment plates 121, the test tubes 15 are clamped in the alignment plates 121, a slide seat 151 is clamped at the bottom of each test tube 15, the slide seat 151 can drive the test tubes 15 to slide in the alignment plates 121, so that the weights of the alignment plates 121 at two ends are kept equal,

wherein, the weight ratio of the alignment plate 121 can be measured by the weight sensor 122, and the slide 151 can drive the test tube 15 to slide left and right on the alignment plate 121, adjust the weight ratio of the two ends, so that the alignment plates 121 at the two ends are balanced, if the weights of the alignment plates 121 at the two ends are unbalanced, then when shaking the bottle, the flora in the test tube 15 is unstable in oscillation, and may overflow from the test tube 15 in serious cases, causing loss, and it needs to be particularly emphasized here that if the test tube 15 cannot be balanced by the alignment plates 121 at the two ends all the time after moving, sterile water can be injected through the empty test tube 15, and the two ends reach symmetrical weights through the cooperation of the sterile water.

Further, the width of the alignment plate 121 can be adjusted, and after the weight is leveled, the test tube 15 can be clamped to follow the universal spring seat 11 for shaking the bottle.

The flora detection device further comprises a flora detection structure 13, the flora detection structure 13 is arranged on the outer surfaces of all test tubes 15 in a surrounding manner, camera assemblies 131 are arranged on the outermost periphery of the flora detection structure 13 at positions opposite to the test tubes 15, a main camera 1312 and an auxiliary camera 1311 are arranged on the camera assemblies 131, the main camera 1312 is opposite to the test tubes 15, the auxiliary cameras 1311 are arranged on two sides of the main camera 1312, the auxiliary cameras 1311 are adjustable in position and are opposite to the test tubes 15,

the test tube 15 can be photographed and sampled, the photographed picture is compared with a normal strain picture in a control system, if the similarity exceeds more than 5%, the test tube is judged to be infected, the photographing and sampling time period is 15min, namely, all the test tubes 15 are photographed and sampled after every 15min, and the quality of the shake flask strain in the obtained test tube 15 is higher.

Further, the interior test tube 15 is relative with more than two vice camera 1311 at least, because the test tube 15 of interior circumference can be blocked by the test tube 15 in the place ahead, can only shoot the mode of sample through vice camera 1311, if only adopt the mode of single vice camera 1311 sample, the error can appear in the inevitable, so, make the test tube 15 of interior circumference can shoot the sample twice at least, through twice contrast, confirm whether infected, make the bacterial can be in monitor state constantly at the in-process of fermentation reproduction, in case take place to stop after infecting rapidly, prevent that one test tube 15 from infecting other bacterial after infecting outside thallus, improve the stability of mushroom liquid bacterial.

Further, a stirring rod 31 is arranged on the second-stage tank 3, a conveying cavity 33 is further arranged at the bottom of the second-stage tank 3, a plurality of permeation tubes 32 are arranged on the conveying cavity 33, specifically, a plurality of stirring branch rods 311 are connected to the stirring rod 31, a plurality of ring pieces 322 are arranged on the permeation tubes 32, a plurality of concave holes 321 are arranged between the ring pieces 322, after the pressure is applied to the ring pieces 322, the concave holes 321 can spray gas into the second-stage tank 3, and the stirring branch rods 311 and the ring pieces 322 are arranged oppositely,

wherein, the infiltration pipe 32 dispersion sets up in whole second grade jar 3, make oxygen can permeate one-level fermentation bacterial from a plurality of positions, it rotates to divide pole 311 to drive the stirring when puddler 31, the stirring divides pole 311 to stir liquid, make the zymotic fluid toward outer whipping, the zymotic fluid of toward outer whipping can extrude ring piece 322, make ring piece 322 to the infiltration pipe 32 internal reaction, outwards spray oxygen through shrinkage pool 321, let oxygen can supply thereupon along with the speed of stirring, in the very fast time cycle of hyperplasia, can improve the volume of oxygen suppliment automatically, in the time cycle that the appreciation is slow, can reduce the volume of oxygen suppliment automatically, the intelligent regulation supplies oxygen volume, make the bacterial strain can reach the biggest bacterial colony, activity when the hyperplasia is higher.

Further, the conveying cavity 33 comprises an upper conductive layer 333, a lower conductive layer 332 is connected to the bottom of the upper conductive layer 333, a sterile connector 331 for connecting an external air source is arranged on the lower conductive layer 332,

wherein, the external gas source is connected with the aseptic connector 331, the introduced oxygen is isolated in the lower conductive layer 332, and after the sterilization operation of the lower conductive layer 3321-2h, the oxygen is introduced into the upper conductive layer 333, wherein the upper conductive layer 333 and the lower conductive layer 332 are in a closed state under a normal state, and the upper conductive layer 333 is connected with the lower conductive layer 332 only after the lower conductive layer 332 is confirmed to be in an aseptic state.

s1; placing the culture medium into the test tube 15, placing the cut mushroom slant strains into the test tube 15, sealing the test tube 15, placing the container into the oscillation box 1,

s2; after the oscillating box 1 is closed, the weight of the alignment plates 121 in the oscillating box 1 is matched through control equipment, so that the weights of the alignment plates 121 at the two ends are consistent,

s3; adjusting the temperature of the oscillation box 1, standing for 40-50h at 25 +/-2 ℃, adjusting the temperature to 17-21 ℃, starting the oscillation box 1, and starting oscillation culture for 60-72h to obtain shake flask strains;

s4; the shake flask strains in the test tube 15 are all extracted through the communicating frame 15, are not contacted with the outside in the extracting process, are directly extracted into the primary tank 2, are sterilized for 2 to 3 hours under the conditions that the steam pressure is 0.15 to 0.19MPa and the temperature is 120 ℃ and 122 ℃ to obtain primary fermentation strains,

s5; introducing a primary fermentation strain into the secondary tank 3, stirring by a stirring rod 31, and continuously supplying oxygen to the primary fermentation strain by a permeation tube 32 in the stirring process, wherein the oxygen introduction amount is 150 +/-10 m3/h, the fermentation temperature is 20 +/-2 ℃, and the tank pressure is 0.15 +/-0.02 MPa, culturing under the conditions of 90-100 to obtain the mushroom liquid strain, preferably, in the stirring process of the primary fermentation strain, the oxygen introduction amount is 155m3/h, the fermentation temperature is 21 ℃, and the tank pressure is 0.17MPa, under the oxygen conditions, the temperature and the pressure, the speed and the activity of strain propagation reach the best, and the culture time is controlled to be 95-96h, so that the strain reaches the maximum flora, if the temperature is high, the water shortage phenomenon easily occurs, the flora is difficult to propagate, if the temperature is too low, the strain activity is low, and the intercellular division is difficult to normally occur.

Further, in step S2, the aligning plates 121 at the left and right ends are respectively empty of one test tube 15, and the communicating rack 15 can fill water into the test tube 15 to balance the weight.

S21; when water is injected, sterile water is not needed.

The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A high activity, high stability mushroom liquid bacterial fermentation system, its characterized in that includes: an oscillating box (1), a primary tank (2) and a secondary tank (3), wherein a liquid outlet of the oscillating box (1) is connected with the primary tank (2), a liquid outlet of the primary tank (2) is connected with the secondary tank (3),

oscillation box (1) includes omnipotent spring holder (11), the top of omnipotent spring holder (11) is provided with balanced splint (12), the top of balanced splint (12) is fixed with a plurality of test tubes (15), test tube (15) can slide on balanced splint (12), and have at least one leave the sky in test tube (15), all the top of test tube (15) all communicates through intercommunication frame (15), intercommunication frame (15) with the inlet intercommunication of one-level jar (2).

2. The high-activity and high-stability mushroom liquid spawn fermentation system according to claim 1, wherein the balance clamp plate (12) is divided into two alignment plates (121) which are bilaterally symmetrical, each alignment plate (121) is internally provided with a weight sensor (122), a plurality of test tubes (15) are clamped in the alignment plates (121), a slide seat (151) is clamped at the bottom of each test tube (15), and the slide seat (151) can drive the test tubes (15) to slide in the alignment plates (121) to keep the weights of the alignment plates (121) at two ends equal.

3. The high-activity and high-stability shiitake mushroom liquid spawn fermentation system according to claim 2, wherein the alignment plate (121) is adjustable in width, and after the weight is leveled, the test tube (15) can be clamped to follow the universal spring seat (11) for shaking the bottle.

4. The system for fermenting the liquid spawn of the lentinus edodes according to claim 1, further comprising a flora detection structure (13), wherein the flora detection structure (13) is arranged around the outer surfaces of all test tubes (15), the flora detection structure (13) is provided with a camera assembly (131) at the relative position of the outermost test tube (15), the camera assembly (131) is provided with a main camera (1312) and an auxiliary camera (1311), the main camera (1312) is opposite to the test tube (15), the auxiliary camera (1311) is arranged at two sides of the main camera (1312), and the auxiliary camera (1311) is adjustable in position and opposite to the test tube (15).

5. The fermentation system of mushroom liquid spawn according to claim 2, 3 or 4, wherein the test tube (15) is opposite to at least two of the auxiliary cameras (1311).

6. The high-activity and high-stability lentinus edodes liquid strain fermentation system as claimed in claim 1, wherein a stirring rod (31) is arranged on said secondary tank (3), a conveying cavity (33) is further arranged at the bottom of said secondary tank (3), and a plurality of permeation tubes (32) are arranged on said conveying cavity (33).

7. The high-activity and high-stability lentinus edodes liquid strain fermentation system as claimed in claim 6, wherein a plurality of stirring branch rods (311) are connected to the stirring rod (31), a plurality of ring sheets (322) are arranged on the permeation tube (32), a plurality of concave holes (321) are arranged between the ring sheets (322), after the ring sheets (322) are pressurized, the concave holes (321) can eject gas into the secondary tank (3), and the stirring branch rods (311) and the ring sheets (322) are arranged oppositely.

8. The high-activity and high-stability mushroom liquid spawn fermentation system according to claim 6, wherein the conveying cavity (33) comprises an upper conducting layer (333), a lower conducting layer (332) is connected to the bottom of the upper conducting layer (333), and a sterile connector (331) for connecting an external gas source is arranged on the lower conducting layer (332).

9. The fermentation method of high activity and high stability mushroom liquid spawn according to claims 1 to 8, characterized by comprising the following steps:

s1; placing the culture medium into the test tube (15), placing the cut mushroom slant strains into the test tube (15), sealing the test tube (15), placing the container into the oscillation box (1),

s2; after the oscillating box (1) is closed, the weight of the alignment plates (121) in the oscillating box (1) is matched through control equipment, so that the weights of the alignment plates (121) at two ends are consistent,

s3; adjusting the temperature of the oscillation box (1), standing for 40-50h at 25 +/-2 ℃, adjusting the temperature to 17-21 ℃, starting the oscillation box (1), and starting oscillation culture for 60-72h to obtain shake flask strains;

s4; the shake flask strains in the test tube (15) are all extracted through the communicating frame (15) without contacting with the outside in the extraction process, and are directly extracted into the primary tank (2), and are sterilized for 2 to 3 hours under the conditions of vapor pressure of 0.15 to 0.19MPa and temperature of 120 to 122 ℃ to obtain primary fermentation strains,

s5; introducing a primary fermentation strain into the secondary tank (3), stirring by a stirring rod (31), continuously supplying oxygen to the primary fermentation strain by a permeation tube (32) in the stirring process, wherein the oxygen supply amount is 150 +/-10 m3/h, the fermentation temperature is 20 +/-2 ℃, and the tank pressure is 0.15 +/-0.02 MPa, and culturing under the condition of 90-100 ℃ to obtain the mushroom liquid strain.

10. The fermentation method of liquid spawn of Lentinus edodes according to claim 9, wherein in step S2, one test tube (15) is left empty in each of the alignment plates (121) at the left and right ends, and the communication rack (15) is used to fill water and balance the test tube (15).