CN114507588A - A kind of inoculation device and inoculation method for producing Monascus red by liquid submerged fermentation of Monascus - Google Patents

Abstract

The invention relates to the technical field of Monascus inoculation, and discloses an inoculation device and an inoculation method for producing Monascus red by liquid deep fermentation of Monascus. An emptying pipe is connected to the upper part, a first regulating valve is arranged on the inlet pipe, a second regulating valve is arranged on the over-type joint, a third regulating valve is arranged on the outlet pipe, and a fourth regulating valve is arranged on the emptying pipe, through which the inlet pipe can go to The seed tank is filled with steam for sterilization, and sterile air can be introduced into the seed tank through the air inlet pipe to maintain a certain positive pressure in the seed tank, and then the inoculation bottle and the seeding joint are connected through the connecting pipe, and the second regulating valve is opened during inoculation. , so that the seed tank and the inoculation bottle are connected so that the pressure is consistent, and then the fourth regulating valve is opened. Inoculation, greatly reducing the infection rate.

Description

A kind of inoculation device and inoculation method for producing Monascus red by liquid submerged fermentation of Monascus

technical field

The invention relates to the technical field of Monascus inoculation, in particular to an inoculation device and an inoculation method for producing Monascus red by liquid submerged fermentation of Monascus.

Background technique

Monascus red refers to the liquid Monascus pigment obtained by extracting red yeast rice with ethanol or the product obtained by extraction, crystallization and purification from the deep culture medium of Monascus. Monascus red pigment is a traditional natural functional food pigment produced by the fermentation of Monascus in my country. Monascus red is a natural functional food coloring produced by microbial fermentation, and it is one of the secondary metabolites of Monascus. Monascus red is a pure natural, non-toxic, safe and reliable natural powder obtained by using rice and soybeans as the main raw materials, drawing on the traditional red yeast production process in my country, through the liquid submerged fermentation of Monascus and advanced modern biological separation technology. food coloring. Monascus red pigment is widely used in the coloring of sake, fermented bean curd, fish meat, soy sauce, cakes, candy, cold drinks and other food industries, and can also be used in the coloring of medicine and cosmetics. Therefore, it is worthy of vigorous application and promotion. Compared with other natural pigments, Monascus red pigment has the advantages of high purity, low price, strong tinting strength, good stability and antiseptic function, so it has great market competitiveness and very important practical significance.

Monascus red is mainly obtained by liquid submerged fermentation of Monascus. In the production of Monascus red by liquid submerged fermentation of Monascus, it is a very critical link to connect the seed solution of the shaker to the seed tank. Because the inoculation environment is a large production workshop rather than a sterile room, it is easy to be affected by the workshop during the inoculation process. Effects of dust and miscellaneous bacteria in the environment. If the bacteria are infected during the inoculation process, it will not only cause huge losses to production, but also affect the shelf life of monascus red products. No matter what kind of inoculation device or inoculation method is used, the most important thing is to greatly reduce the infection rate caused by inoculation. At present, the main inoculation method is to set a fire ring at the mouth of the seed tank. Under the protection of the fire ring, the seed liquid in the inoculation bottle is poured into the seed tank. When the seed tank and the inoculation bottle are opened, the air enters the seed tank or the inoculation bottle. , the infection rate is high. In addition, there is also a method of transferring the bacterial liquid by connecting the inoculation bottle and the seed tank with a tube. Since Monascus is in the shape of a filamentous strip, the seed liquid is thick and easy to block.

The prior art discloses an inoculation device, which includes an inoculation bottle, a bottle cap connected to the bottle mouth at the top of the inoculation bottle through a thread, and an inoculation valve connected to the inoculation mouth at the bottom of the inoculation bottle. There is a silicone gasket. When the bottle cap is tightened, the silicone gasket is used to seal the bottle mouth of the inoculation bottle. The bottle cap is provided with a vent hole, and the vent hole is located near the lower edge of the silicone gasket. This patent performs steam sterilization on the inoculation device during inoculation and steam sterilization of the seed pot at the same time. Open the inoculation valve, unscrew the cap on the inoculation bottle by about two turns, so that there is a gap between the silicone gasket in the bottle cap and the bottle mouth of the inoculation bottle, so that the steam is discharged from the exhaust hole on the bottle cap to ensure the flow of steam Unobstructed, leaving no sterilization dead ends. When the sterilization is completed, the inoculation valve is open, so that the air in the seed tank is filled with the inoculation bottle, and at the same time, the gap between the inoculation bottle and the silicone gasket in the bottle cap is tightened a little more, about one turn, and the air is exhausted to ensure that the inoculation bottle is filled with air. The pressure is positive pressure. Before inoculation, the cotton in the closed fire ring was filled with 95% ethanol. When inoculating, insert the booster rod into the movable pull ring on the bottle cap, put the circular groove of the closed inoculation fire ring on the inoculation bottle, light the fire ring, and close the inoculation valve. Unscrew the bottle cap with an afterburner, place the bottle cap above the flame, and then pour the seed solution in the seed mother bottle into the inoculation bottle under the flame. After pouring the seed solution, tighten the cap on the inoculation bottle with an afterburner rod to ensure that the inoculation bottle and the silicone gasket in the cap have no gaps and no air leakage. At the same time, open the inoculation valve. At this time, the pressure in the inoculation bottle is equal to that in the seed tank. The seed liquid in the inoculation bottle naturally flows into the seed tank, and the inoculation valve is opened. Remove the closed inoculation fire ring, and observe whether the seed liquid flows into the seed tank from the sight glass of the seed tank. After the seed liquid has completely flowed into the seed tank, close the inoculation valve and the inoculation operation is completed. The inoculation bottle of this patent is connected with the seed tank. The function of the inoculation bottle is to slowly add the seed liquid into the seed tank. The inoculation bottle does not play the role of transferring the seed liquid. Pour the seed liquid in the seed bottle into the inoculation bottle under the protection of The seed liquid of the bacteria is thick, which makes the tank mouth easy to block. Therefore, it is difficult for the seed liquid in the inoculation bottle to enter the seed tank.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide an inoculation device and an inoculation method for producing Monascus red by liquid submerged fermentation of Monascus with high efficiency and low bacterial infection rate.

In order to achieve the above purpose, the present invention provides an inoculation device for producing Monascus red by liquid submerged fermentation of Monascus, including an inoculation bottle and a seed tank, the inoculation bottle comprising an inoculation bottle body, a bottle stopper and a connecting pipe, the inoculation bottle The top of the bottle body is provided with a bottle mouth, the bottle stopper is used to plug into the bottle mouth, the side wall of the inoculation bottle body is provided with a connecting mouth, and one end of the connecting pipe is connected with the connecting mouth, and the The other end of the connecting pipe is provided with a seed outlet; the seed tank includes a seed tank body, an air inlet pipe, a seed feeding joint, an air outlet pipe and an emptying pipe, one end of the air inlet pipe is connected with the seed tank body, and the The other end of the air inlet pipe is used to connect with the steam generator and the sterile air compressor. The air inlet pipe is provided with a first regulating valve. One end of the seed passing pipe is connected with the seed tank body, the other end of the seed passing pipe is connected with the joint mouth, the joint mouth cover is threadedly connected with the joint mouth to cover the joint mouth, and the joint mouth is covered by the joint mouth. The seed passage is provided with a second regulating valve, one end of the air outlet is connected with the seed tank, the other end of the air outlet is connected with a third regulating valve, and the third regulating valve is connected with a flow meter, so The air outlet pipe is connected with a pressure gauge, the emptying pipe is connected with the air outlet pipe, and a fourth regulating valve is arranged on the emptying pipe.

As a preferred solution, the air inlet pipe is connected to the bottom end of the seed tank body, and the connection between the seed passage pipe and the seed tank body and the connection point between the air outlet pipe and the seed tank body are located at the the top of the seed tank.

As a preferred solution, the inoculation bottle further includes a fixing jacket, the fixing jacket includes a first splint, a second splint and a locking bolt, the first splint is located above the bottle stopper, and the second splint Sleeved at the mouth of the bottle, the first splint is provided with two symmetrically distributed first through holes, the second splint is provided with two symmetrically distributed second through holes, and the second through holes are A locking nut is provided, and the locking bolt passes through the first through hole and is connected with the locking nut.

As a preferred solution, the joint nozzle includes a fixing part and a connecting part, the fixing part and the connecting part are both cylindrical, and the diameter of the fixing part is larger than the diameter of the connecting part, and one end of the connecting part is It is coaxially and fixedly connected with one end of the fixed part, the fixed part is connected with the end of the seeding pipe, the inner wall of the seeding head is provided with a first internal thread, and the outer wall of the connecting part is provided with a first internal thread. The first external thread matched with the first internal thread, the inner wall of the joint nozzle cover is provided with a second internal thread, and the outer wall of the fixing portion is provided with a second external thread matched with the second internal thread .

The present invention also provides an inoculation method for producing Monascus red by liquid submerged fermentation of Monascus, comprising the following steps:

Step 1: After adding culture medium to the seed tank, carry out actual elimination;

Step 2: After the actual elimination, sterile air is introduced into the seed tank through the air inlet pipe, and the first regulating valve and the third regulating valve are adjusted to maintain a certain positive pressure in the seed tank. The positive pressure is ≥ 0.08MPa and the positive pressure <0.1MPa, and keep the pressure;

Step 3: Sterilize the inoculation bottle, pour the seed liquid of the shaker bottle into the inoculation bottle body using aseptic operation technology in the sterile room, plug the bottle mouth of the inoculation bottle body with a cork, and then use a fixed jacket. Tighten the bottle stopper, wrap the connecting tube around the inoculation bottle, and take it to the workshop for inoculation;

Step 4: Under the protection of the fire ring, remove the cover of the joint mouth, and quickly remove the protective layer wrapped around the seed head, and connect the seed head with the joint mouth; after the seed head and the joint mouth are connected, Fully open the second regulating valve to make the inoculation bottle communicate with the seed tank and have the same pressure; remove the fire ring;

Step 5: During inoculation, control the pressure in the seed tank to be no more than 0.1MPa, slowly open the fourth regulating valve, and the gas in the seed tank will be discharged, so that the pressure in the seed tank will drop, and the seed liquid in the inoculation tank will be pressurized. Into the seed tank, wherein, when the seed liquid in the inoculation bottle is transferred to the seed tank, the fourth regulating valve is not fully opened, and by adjusting the fourth regulating valve, the seed liquid is transferred multiple times until the inoculation bottle is in the body. The seed liquid is all transferred to the seed tank;

Step 6: Disconnect the connection between the seed head and the joint mouth, half-open the second regulating valve, a small amount of mycelium is sprayed out of the joint mouth, clean the joint mouth, close the second regulating valve, and cover the joint mouth on the joint mouth Cover, inoculation is complete.

As a preferred solution, in step 1, during the actual elimination, steam is introduced into the seed tank through the air inlet pipe. At this time, the first regulating valve, the third regulating valve and the fourth regulating valve are opened, and when the temperature in the seed tank rises When the temperature reaches 121°C and the pressure gauge is 0.11MPa, fully open the second regulating valve, and unscrew the joint nozzle cover, the steam is discharged from the joint nozzle, then connect the joint nozzle cover to the joint nozzle, and control the tightening of the joint nozzle cover After that, the temperature in the seed tank is kept at 121-124°C for 30-35 minutes. Finally, the second regulating valve is kept fully open, the joint nozzle cover is tightened, and the fourth regulating valve is closed. .

As a preferred solution, in step 3, when the inoculation bottle is sterilized, after the bottle stopper is tightly closed to the bottle mouth of the inoculation bottle body, the top of the inoculation bottle body is wrapped with kraft paper, the kraft paper wraps the bottle stopper, and then the fixing clip The sleeve is wrapped with kraft paper alone, the seed head is wrapped with kraft paper, the connecting tube is wrapped around the inoculation bottle body, the inoculation bottle and the fixed jacket wrapped with kraft paper are placed in a horizontal sterilization pot, and the temperature is maintained at 121 ~ 124 ℃, Sterilize for 30 min, and after sterilization, take out the inoculation bottle and place it in a sterile room to cool for use.

As a preferred solution, in step 4, the operator wears fire-resistant and heat-insulating gloves to remove the joint mouth cover, and after the seed head and the joint mouth are connected, wet the fire-resistant heat-insulating gloves and wrap the seed head and joint. the connection of the mouth, so that it cools quickly.

As a preferred solution, in step 4, after the seed head is connected with the joint mouth, a transparent protective cover is provided on the outer casing of the inoculation bottle body, and the protective cover is not higher than the connecting mouth.

As a preferred solution, in step 5, while opening the fourth regulating valve, tilt the inoculation bottle body to make the seed liquid in the inoculation bottle close to the connecting mouth; when the pressure gauge drops to 0.02MPa, immediately close the fourth regulating valve to make The pressure in the seed tank rises to 0.08-0.1MPa. During the process of pressure recovery, the inoculation bottle is upright, so that the seed liquid leaves the connecting nozzle, and the seed liquid in the connecting pipe returns to the inoculation bottle; Slowly open the fourth regulating valve, and cycle in this way until all the seed liquid in the inoculation bottle is transferred to the seed tank.

Compared with the prior art, the beneficial effects of the present invention are:

In the inoculation device of the present invention, the seed tank body is connected with a seeding joint, an air inlet pipe and an air outlet pipe; The air outlet pipe is provided with a third regulating valve, and the evacuation pipe is provided with a fourth regulating valve. Through the air inlet pipe, steam can be filled into the seed tank for sterilization. The whole seed tank can be sterilized by flowing out from the emptying pipe, and sterile air can be introduced into the seed tank through the air inlet pipe to maintain a certain positive pressure in the seed tank, and then the inoculation bottle and the filter can be connected through the connecting pipe. Seed joint, the inoculation bottle is poured into the sterile room with the seed liquid, and the seed liquid is moved from the sterile room to the seed tank. During inoculation, the second regulating valve is opened, so that the seed tank and the inoculation bottle are connected and the pressure is consistent. , and then open the fourth regulating valve, the gas in the seed tank is discharged, and the pressure in the seed tank is reduced. , to achieve inoculation. The device of the present invention does not need to sterilize the surrounding environment of the seed tank specially before inoculation, and only needs to keep the workshop in a clean state under normal conditions. Bacteria rate.

In the method of the present invention, during the sterilization process of the seed tank, steam is introduced into the seed tank through the air inlet pipe, and the steam is discharged from the seeding joint, the air outlet pipe and the emptying pipe, so that the whole seed tank can be completely sterilized and sterilized. The effect is good, and sterile air can be introduced into the seed tank through the air inlet pipe, and the first regulating valve and the third regulating valve can be adjusted to maintain a certain positive pressure in the seed tank. The fourth regulating valve is closed, then close the second regulating valve under the protection of the fire ring, and then connect the inoculation bottle and the seed tank to avoid contamination of the joint, and then open the second regulating valve to make the inoculation bottle and the seed tank communicate with each other. In order to make the two pressures the same, then open the fourth regulating valve to reduce the pressure in the seed tank, so that the seed liquid in the inoculation bottle is pressed into the seed tank, and a small amount of transfer method is adopted during inoculation, which can reduce the pressure in the seed tank. It avoids the problems of clogging the connecting nozzle and preventing the rupture of the inoculation bottle, and has high inoculation efficiency and low bacterial infection rate.

Description of drawings

1 is a schematic structural diagram of an inoculation device for producing Monascus red by liquid submerged fermentation of Monascus in accordance with an embodiment of the present invention.

FIG. 2 is a schematic structural diagram of an inoculation bottle according to an embodiment of the present invention.

FIG. 3 is a schematic structural diagram of a seed pot according to an embodiment of the present invention.

Fig. 4 is the flow chart of the inoculation method of Monascus liquid submerged fermentation to produce Monascus red according to the embodiment of the present invention.

In the figure, 1-inoculation bottle body; 2-bottle stopper; 3-connecting pipe; 4-connecting nozzle; 5-seed head; 6-seed tank; 7-inlet pipe; 8-outlet pipe; 9-empty pipe; 10-first regulating valve; 11-seeding pipe; 12-joint nozzle; 1201-fixing part; 1202-connecting part; 13-joint nozzle cover; 14-second regulating valve; 15-third regulating valve; 16-flow meter; 17-pressure gauge; 18-fourth regulating valve; 19-first splint; 20-second splint; 21-locking bolt; 22-protective sleeve; 23-stirring motor; 24-rotating shaft; 25 - Stirring blades.

Detailed ways

The specific embodiments of the present invention will be described in further detail below with reference to the accompanying drawings and embodiments. The following examples are intended to illustrate the present invention, but not to limit the scope of the present invention.

In the description of the present invention, it should be noted that the terms "center", "portrait", "horizontal", "top", "bottom", "front", "rear", "left", "right", " The orientation or positional relationship indicated by vertical, horizontal, top, bottom, inner, outer, etc. is based on the orientation or positional relationship shown in the drawings, and is only for the convenience of describing the present invention and The description is simplified rather than indicating or implying that the device or element referred to must have a particular orientation, be constructed and operate in a particular orientation, and therefore should not be construed as limiting the invention. Furthermore, the terms "first", "second", and "third" are used for descriptive purposes only and should not be construed to indicate or imply relative importance.

In the description of the present invention, it should be noted that the terms "installed", "connected" and "connected" should be understood in a broad sense, unless otherwise expressly specified and limited, for example, it may be a fixed connection or a detachable connection Connection, or integral connection; can be mechanical connection, can also be electrical connection; can be directly connected, can also be indirectly connected through an intermediate medium, can be internal communication between two elements. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood in specific situations.

Example 1

As shown in Figure 1 to Figure 3, a preferred embodiment of the present invention is a kind of inoculation device for producing Monascus red by liquid submerged fermentation of Monascus, including an inoculation bottle and a seed tank, and the inoculation bottle includes an inoculation bottle body 1, a bottle stopper 2 and The connecting pipe 3, the top of the inoculation bottle body 1 is provided with a bottle mouth, the bottle stopper 2 is used to plug the bottle mouth, the side wall of the inoculation bottle body 1 is provided with a connecting nozzle 4, and one end of the connecting pipe 3 is connected with the connecting mouth 4, and the connection The other end of the pipe 3 is provided with a seed head 5; the seed tank includes a seed tank body 6, an air inlet pipe 7, a seeding joint, an air outlet pipe 8 and an emptying pipe 9, and one end of the air inlet pipe 7 is connected with the seed tank body 6, and the inlet pipe 7 is connected to the seed tank body 6. The other end of the trachea 7 is used to connect with the steam generator and the sterile air compressor. The air inlet pipe 7 is provided with a first regulating valve 10. The seeding joint includes a seeding pipe 11, a joint nozzle 12 and a joint nozzle cover 13. One end of the seed tube 11 is connected with the seed tank body 6 , the other end of the seed tube 11 is connected with the joint nozzle 12 , the joint nozzle cover 13 is threadedly connected with the joint nozzle 12 to cover the joint nozzle 12 , and the seed tube 11 is provided with a second The regulating valve 14, one end of the gas outlet pipe 8 is connected with the seed tank 6, the other end of the gas outlet pipe 8 is connected with a third regulating valve 15, the third regulating valve 15 is connected with a flow meter 16, and the gas outlet pipe 8 is connected with a pressure gauge 17, The emptying pipe 9 is connected with the air outlet pipe 8 , and a fourth regulating valve 18 is arranged on the emptying pipe 9 . In the inoculation device of the present embodiment, the seed tank body 6 is connected with a seeding joint, an air inlet pipe 7 and an air outlet pipe 8. The air outlet pipe 8 is connected with an emptying pipe 9, and the air inlet pipe 7 is provided with a first regulating valve 10. The joint is provided with a second regulating valve 14, the air outlet pipe 8 is provided with a third regulating valve 15, and the exhaust pipe 9 is provided with a fourth regulating valve 18. Through the air inlet pipe 7, the seed tank 6 can be filled with steam for sterilization. And in the sterilization process, the steam flows out from the seed joint, the air outlet pipe 8 and the emptying pipe 9, which can sterilize the seed tank as a whole, and can pass sterile air into the seed tank body 6 through the air inlet pipe 7. In order to maintain a certain positive pressure in the seed tank body 6, the inoculation bottle and the seeding joint are connected through the connecting pipe 3, the seed liquid is poured into the inoculation bottle in the sterile room, and the seed liquid is moved from the sterile room to the seed tank. , During inoculation, open the second regulating valve 14 to make the seed tank 6 communicate with the inoculation bottle 1 so that the pressure is consistent, and then open the fourth regulating valve 18, the gas in the seed tank 6 is discharged, so that the seed tank 6 The internal pressure is reduced, and under the action of the pressure difference, the seed liquid flows from the inoculation bottle 1 through the connecting pipe 3 and the seeding joint into the seed tank 6 to realize inoculation. Before inoculation, the device of this embodiment does not need to sterilize the surrounding environment of the seed tank, and it is enough to keep the workshop in a clean state. Small contamination rate.

In this embodiment, the air inlet pipe 7 is connected to the bottom end of the seed tank 6 , and the connection between the seed pipe 11 and the seed tank 6 and the connection between the air outlet pipe 8 and the seed tank 6 are located at the top of the seed tank 6 . The steam enters from the bottom of the seed tank body 6 and flows out from the top, so that the steam can flow through the whole of the seed tank body 6 to achieve comprehensive sterilization. And during inoculation, sterile air will pass through the liquid in the seed tank 6, and the liquid will be turned over to make the medium and the seed liquid evenly mixed.

Optionally, the bottle stopper 2 in this embodiment is a rubber stopper. The connecting pipe 3 is a silicone tube with high temperature resistance, high pressure resistance and corrosion resistance. The inner diameter of the connecting pipe 3 in this embodiment is 12 mm. The inoculation bottle body 1 is a transparent body, which can observe the internal situation of the inoculation bottle body 1, monitor the process of the flow of the seed liquid, and see the flow of the seed liquid during inoculation, and determine whether the seeding is completed. The seed tank body 6 is a non-transparent body, and a sight glass is provided on the seed tank body 6 to observe the internal situation. The inoculation bottle body 1 of this embodiment is preferably a vacuum suction filter bottle.

In addition, the first regulating valve 10 and the third regulating valve 15 in this embodiment are diaphragm valves, and the second regulating valve 14 and the fourth regulating valve 18 are ball valves. The flow meter 16 is a rotameter, which is used to monitor the air output of the third regulating valve 15 , so that the first regulating valve 10 and the third regulating valve 15 cooperate to control the pressure in the seed tank 6 . The pressure gauge 17 shows the pressure in the seed tank 6. During inoculation, the pressure in the seed tank 6 can be monitored at all times to prevent the inoculation tank 1 from bursting due to excessive pressure, and to prevent the pressure in the seed tank 6 from falling to 0.

Embodiment 2

The difference between this embodiment and the first embodiment is that, on the basis of the first embodiment, this embodiment further describes the structures of the inoculation bottle and the joint nozzle 12 .

In this embodiment, the inoculation bottle further includes a fixed jacket, the fixed jacket includes a first splint 19, a second splint 20 and a locking bolt 21, the first splint 19 is located above the bottle stopper 2, and the second splint 20 is sleeved At the mouth of the bottle, the first splint 19 is provided with two symmetrically distributed first through holes, the second splint 20 is provided with two symmetrically distributed second through holes, and a locking nut is provided in the second through hole for locking. The bolt 21 passes through the first through hole and is connected with the lock nut. By rotating the locking nut 21, the first splint 19 and the second splint 20 can be brought closer, so that the bottle stopper 2 is pressed tightly, and the sealing performance is good, so as to avoid contamination.

In addition, the joint nozzle 12 of this embodiment includes a fixing portion 1201 and a connecting portion 1202. Both the fixing portion 1201 and the connecting portion 1202 are cylindrical, and the diameter of the fixing portion 1201 is larger than the diameter of the connecting portion 1202. One end of the connecting portion 1202 is connected to the fixing portion 1202. One end of the part 1201 is coaxially and fixedly connected, the fixed part 1201 is connected with the end of the seed pipe 11, the inner wall of the seed head 5 is provided with a first internal thread, and the outer wall of the connecting part 1202 is provided with a first internal thread. The first external thread, the inner wall of the joint nozzle cover 13 is provided with a second internal thread, and the outer wall of the fixing portion 1201 is provided with a second external thread matched with the second internal thread. The seed head 5 can be screwed into the connection part 1202 of the joint nozzle 12, and the joint nozzle cover 13 can be screwed onto the fixed part 1201 of the joint nozzle 12, which is convenient to connect and has good sealing performance. The joint nozzle cover 13 is screwed tightly with the joint nozzle 12 after the actual elimination, which can prevent the joint nozzle 12 from being polluted.

In addition, the inoculation bottle of this embodiment also includes a protective cover 22, which is set outside the inoculation bottle body 1. When the inoculation bottle body 1 is accidentally broken, the protective cover 22 can prevent debris and seed liquid from splashing, and protects effect. In addition, the protective cover 22 in this embodiment is a transparent bag body, so that the amount of the seed liquid in the inoculated bottle body 1 can be observed.

The seed tank of this embodiment also includes a stirring device, which includes a stirring motor 23, a rotating shaft 24 and a stirring blade 25. The stirring motor 23 is located outside the seed tank 6, one end of the rotating shaft 24 is connected to the stirring motor 23, and the other end of the rotating shaft 24 is connected to the stirring motor 23. The top of the seed tank body 6 extends into the seed tank body 6 , and the stirring blade 25 is connected to the rotating shaft 24 . The stirring motor 23 drives the rotating shaft 24 to rotate, and the stirring blade 25 connected to the rotating shaft 24 rotates with the rotating shaft 24 to realize stirring, so that the seed liquid flowing into the seed tank 6 and the culture medium in the seed tank 6 are evenly mixed.

Other structures of this embodiment are the same as those of the first embodiment, and are not repeated here.

Embodiment 3

As shown in Figure 4, the inoculation method of a kind of Monascus liquid submerged fermentation to produce Monascus red according to a preferred embodiment of the present invention, comprises the steps:

Step 1: After adding the culture medium in the seed tank 6, carry out actual elimination;

Step 2: After the actual elimination, sterile air is introduced into the seed tank 6 through the air inlet pipe 7, and the first regulating valve 10 and the third regulating valve 15 are adjusted to maintain a certain positive pressure in the seed tank 6, and the positive pressure ≥ 0.08MPa and the positive pressure is less than 0.1MPa, and the pressure is maintained; in this embodiment, after the actual elimination, sterile air is immediately introduced into the seed tank 6 to maintain a certain positive pressure in the seed tank 6 until inoculation. The seed tank body 6 maintains a certain positive pressure during the time between actual elimination and inoculation, to prevent outside air or miscellaneous bacteria from entering the seed tank body 6, and to keep the inside of the seed tank body 6 clean;

Step 3: Sterilize the inoculation bottle, pour the seed liquid of the shaker bottle into the inoculation bottle body 1 using aseptic operation technology in the sterile room, plug the bottle mouth of the inoculation bottle body 1 with the bottle stopper 2, and then use The fixed jacket compresses the bottle stopper 2, wraps the connecting tube 3 on the inoculation bottle body 1, and takes it to the workshop for inoculation;

Step 4: Under the protection of the fire ring, remove the joint nozzle cover 13, and quickly remove the protective layer wrapped by the seed head 5. The protective layer in this embodiment is kraft paper, and the seed head 5 and the joint nozzle 12 are removed. connection; after the seed head 5 and the joint mouth 12 are connected, fully open the second regulating valve 14 to make the inoculation bottle 1 communicate with the seed tank 6 and have the same pressure; remove the fire ring; The fire ring is removed after the regulating valve 14 for a period of time, so that the fire ring continues to protect the connection between the seed head 5 and the joint mouth 12 for a period of time;

Step 5: During inoculation, control the maximum pressure in the seed tank 6 to not exceed 0.1 MPa, slowly open the fourth regulating valve 18, and discharge the gas in the seed tank 6, so that the pressure in the seed tank 6 drops, inoculation. The seed liquid in the bottle body 1 is pressed into the seed tank body 6, wherein, when the seed liquid in the inoculation bottle body 1 is transferred to the seed tank body 6, the fourth regulating valve 18 is not fully opened. Valve 18, so that the seed liquid is transferred multiple times, until all the seed liquid in the inoculation bottle 1 is transferred to the seed tank 6;

Step 6: Disconnect the seed head 5 from the joint nozzle 12, half-open the second regulating valve 14, a small amount of mycelium is ejected from the joint nozzle 12, clean the joint nozzle 12, close the second regulating valve 14, The nozzle 12 is covered with the joint nozzle cover 13, and the inoculation is completed.

After the method of this embodiment is completed, sterile air is introduced into the seed tank body 6 through the air inlet pipe 7, and the first regulating valve 10 and the third regulating valve 15 are adjusted, so that a certain positive pressure can be maintained in the seed tank body 6. , then connect the inoculation bottle and the seed tank under the protection of the fire ring to avoid contamination of bacteria at the connection, and then open the second regulating valve 14 to make the inoculation bottle 1 and the seed tank 6 communicate with each other so that the pressures of the two are consistent, and then open the second regulating valve 14. The four regulating valves 18 reduce the pressure in the seed tank body 6, so that the seed liquid in the inoculation bottle body 1 is pressed into the seed tank body 6, and a small amount of transfer method is adopted during inoculation, which can avoid connecting the nozzle. The problems of clogging and preventing the rupture of the inoculation bottle body 1 have high inoculation efficiency and low bacterial infection rate. The seed liquid in the inoculation bottle 1 flows into the seed tank 6 under the action of the pressure difference, which can realize rapid flow and improve efficiency.

Specifically, in step 1, during the actual elimination, steam is introduced into the seed tank 6 through the air inlet pipe 7, and the steam is saturated steam. At this time, the first regulating valve 10, the third regulating valve 15 and the fourth regulating valve Valve 18 is open. When the temperature in the seed tank 6 rises to 90° C., the fourth regulating valve 18 is closed to half-open, which can reduce the outflow of steam and make the seed tank 6 heat up rapidly. When the temperature in the seed tank 6 rises to 121°C and the pressure gauge is 0.11MPa, fully open the second regulating valve 14 and unscrew the joint nozzle cover 13 , steam is discharged from the joint nozzle 12 , and then the joint nozzle cover 13 It is connected with the joint nozzle 12, and the degree of tightening of the joint nozzle cover 13 is controlled, so that the steam is obviously discharged from the joint nozzle 12. Then, the temperature in the seed tank 6 is maintained at 121-124 ° C for 30-35 minutes, and finally, the first The second regulating valve 14 is kept fully open, the joint nozzle cover 13 is tightened, and the fourth regulating valve 18 is closed. In this embodiment, after the actual elimination is completed, sterile air is immediately introduced to maintain a certain positive pressure in the seed tank body 6 until inoculation. The air is discharged from the joint nozzle 12 to prevent outside air from entering the seed tank body 6 . During the sterilization process of the seed tank, steam is introduced into the seed tank body 6 through the air inlet pipe 7, and the steam is discharged from the seeding joint, the air outlet pipe 8 and the evacuation pipe 9, which can completely sterilize the whole seed tank. Bacterial effect is good.

In step 3, when sterilizing the inoculation bottle, after the bottle stopper 2 is tightly plugged the bottle mouth of the inoculation bottle body 1, the top of the inoculation bottle body 1 is wrapped with kraft paper, the kraft paper wraps the bottle stopper 2, and then the fixing clip The sleeve is wrapped with kraft paper alone, the seed head 5 is wrapped with kraft paper, the connecting tube 3 is wrapped around the inoculation bottle body 1, the inoculation bottle and the fixed jacket wrapped with kraft paper are placed in a horizontal sterilization pot, and the temperature is maintained at 121～ Sterilize at 124°C for 30 min. After sterilization, take out the inoculation bottle and place it in a sterile room to cool for later use. Select the inoculation bottle body 1, and select the volume of the inoculation bottle body 1 according to the volume of the seeds when the bottles are combined. When combining bottles, remove the kraft paper wrapped on the top of the inoculation bottle body 1, remove the bottle stopper 2, and pour the seed solution in the shake flask into the inoculation bottle on the ultra-clean workbench in the sterile room to complete the bottle combination. . After the bottle is combined, plug the bottle stopper 2 tightly, then tear off the kraft paper wrapping the fixed jacket, press the bottle stopper 2 with the fixed jacket, and wrap the connecting tube 3 on the inoculation bottle body 1. Workshop to be inoculated. Combining flasks refers to combining several cultured shake flask seed liquids into inoculation flasks.

In addition, when the connecting pipe 3 of this embodiment is connected with the connecting nozzle 4 and the seeding head 5, first use small iron wires to tie the connection between the connecting pipe 3 and the connecting nozzle 4 and the connecting point between the connecting pipe 3 and the seeding head 5 respectively. Tighten it tightly with another layer of cotton cloth, and then tie the outer layer with kraft paper.

In step 4, three operators are required, one person ignites the fire ring, one person controls the inoculation bottle, and one person controls and monitors the pressure in the seed tank 6 after connecting the inoculation bottle and the seed tank. When connecting the inoculation bottle and the seed pot, the operator wears refractory and heat-insulating gloves to remove the joint nozzle cover 13, and screw the seed head 5 into the joint mouth 12. When screwing the seed head 5, follow the The rotation direction of the head 5 rotates the inoculation bottle body 1 to prevent the connecting tube 3 from being twisted and to avoid affecting the flow of seeds. Since the second regulating valve 14 is fully open during the period from the actual cancellation to the time of inoculation, the second regulating valve 14 is first closed under the protection of the fire ring before removing the joint nozzle cover 13 . Then, under the protection of the fire ring, connect the inoculation bottle and the seed pot to avoid contamination of bacteria at the connection, and then open the second regulating valve 14 . And after the seed head 5 and the joint mouth 12 are connected, wet the fire-resistant heat-insulating gloves and wrap the joint between the seed head 5 and the joint mouth 12 to cool them quickly.

In step 4, after the seed head 5 is connected with the joint nozzle 12, a transparent protective cover 22 is provided on the outer jacket of the inoculation bottle body 1, and the protective cover 22 is not higher than the connecting nozzle 4, so that the protective cover 22 does not cover the connection. Mouth 4. The protective cover 22 can prevent debris and liquid from splashing when the inoculation bottle body 1 is broken, and play a protective role.

In step 5, while opening the fourth regulating valve 18, tilt the inoculation bottle body 1 so that the seed liquid in the inoculation bottle body 1 is close to the connecting mouth 4, but does not completely submerge the connecting mouth 4, so as to prevent the Monascus seeds from being thick and Blockage; the seed liquid is pressed into the seed tank 6 under the action of the pressure difference. When the pressure gauge drops to 0.02MPa, immediately close the fourth regulating valve 18, so that the pressure in the seed tank 6 rises to 0.08-0.1MPa. During the pressure recovery, the inoculation bottle 1 is erected to make the seed liquid leave the connection Mouth 4, the seed liquid in the connecting pipe 3 is returned to the inoculation bottle 1; when the pressure rises, the fourth regulating valve 18 is slowly opened, and this cycle is repeated until all the seed liquid in the inoculation bottle is transferred to the seed tank. . The seed solution in this embodiment is not finished once, but requires multiple operations. During operation, the fourth regulating valve 18 cannot be fully opened to prevent the pressure of the seed tank 6 from dropping to zero.

In step 6, wipe the joint nozzle 12 that sprays out the mycelium with a small cotton cloth with alcohol. The alcohol swab in this example is 75% alcohol.

The seed liquid in the inoculation bottle 1 of this embodiment flows into the seed tank 1 without contact with the external environment. After years of production verification of Monascus red, the infection rate caused by inoculation is less than or equal to 1.0%.

To sum up, the embodiment of the present invention provides an inoculation device for producing Monascus red by liquid submerged fermentation of Monascus, which connects the seed tank body 6 with the seed joint, the air inlet pipe 7 and the air outlet pipe 8, and the air outlet pipe 8 is connected to the inoculation device. There is an emptying pipe 9, the intake pipe 7 is provided with a first regulating valve 10, the over-type joint is provided with a second regulating valve 14, the air outlet pipe 8 is provided with a third regulating valve 15, and the emptying pipe 9 is provided with a fourth regulating valve 18 , through the air inlet pipe 7, the seed tank body 6 can be filled with steam for sterilization, and during the sterilization process, the steam flows out from the seeding joint, the air outlet pipe 8 and the emptying pipe 9, and the whole seed tank can be sterilized. bacteria, and sterile air can be introduced into the seed tank 6 through the air inlet pipe 7 to maintain a certain positive pressure before the inoculation, and then the inoculation bottle and the seeding joint are connected through the connecting pipe 3, and the inoculation bottle is poured into the sterile room. Seed liquid, move the seed liquid from the sterile room to the seed tank, open the second regulating valve 14, make the seed tank 6 communicate with the inoculation bottle 1 so that the pressure is consistent, and then open the fourth regulating valve 18, the seed tank The gas in the 6 is discharged to reduce the pressure in the seed tank 6. Under the action of the pressure difference, the seed liquid flows from the inoculation bottle 1 through the connecting pipe 3 and the seeding joint into the seed tank 6 to realize inoculation. Before inoculation, the device of this embodiment does not need to sterilize the surrounding environment of the seed tank, and it is enough to keep the workshop in a clean state. Small contamination rate. The device of the embodiment of the present invention has firm structure, good sealing performance and simple operation, and is very suitable for Monascus inoculation. In addition, the embodiment of the present invention also provides a kind of inoculation method of Monascus monascus liquid submerged fermentation to produce Monascus red. After the actual elimination, sterile air is introduced into the seed tank 6 through the air inlet pipe 7, and the first regulating valve 10 is adjusted. With the third regulating valve 15, a certain positive pressure can be maintained in the seed tank 6 until inoculation. At this time, the second regulating valve 14 is kept fully open, the fourth regulating valve 18 is closed, and then the third regulating valve is closed under the protection of the fire ring. The second regulating valve 14 is then connected to the inoculation bottle and the seed tank to avoid contamination of bacteria at the connection, and the second regulating valve 14 is opened again to make the inoculation bottle 1 and the seed tank 6 communicate with each other so that the pressures of the two are the same, and then the fourth regulating valve is opened. The valve 18 reduces the pressure in the seed tank body 6, so that the seed liquid in the inoculation bottle body 1 is pressed into the seed tank body 6, and a small amount of transfer method is adopted during inoculation, which can avoid the blockage of the connecting nozzle and the The problem of breaking the inoculation bottle body 1 is prevented, the inoculation efficiency is high, and the bacterial infection rate is low. The seed liquid in the inoculation bottle 1 flows into the seed tank 6 under the action of the pressure difference, which can realize rapid flow and improve efficiency.

The above are only the preferred embodiments of the present invention. It should be pointed out that for those skilled in the art, without departing from the technical principles of the present invention, several improvements and replacements can be made. These improvements and replacements It should also be regarded as the protection scope of the present invention.

Claims (10)

1. a kind of inoculation device of Monascus liquid submerged fermentation to produce Monascus red, is characterized in that, comprises inoculation bottle and seed tank, and described inoculation bottle comprises inoculation bottle body (1), bottle stopper (2) and connecting pipe ( 3), the top of the inoculation bottle body (1) is provided with a bottle mouth, the bottle stopper (2) is used for plugging into the bottle mouth, and the side wall of the inoculation bottle body (1) is provided with a connecting nozzle ( 4), one end of the connecting pipe (3) is connected with the connecting nozzle (4), and the other end of the connecting pipe (3) is provided with a seed head (5); The seed tank comprises a seed tank body (6), an air inlet pipe (7), a seeding joint, an air outlet pipe (8) and an exhaust pipe (9), and one end of the air inlet pipe (7) is connected to the seed tank body (6) connection, the other end of the air inlet pipe (7) is used to connect with a steam generator and a sterile air compressor, the air inlet pipe (7) is provided with a first regulating valve (10), the The seed joint comprises a seed pipe (11), a joint nozzle (12) and a joint nozzle cover (13), one end of the seed pipe (11) is connected to the seed tank body (6), and the seed pipe ( The other end of 11) is connected with the joint nozzle (12), the joint nozzle cover (13) is screwed with the joint nozzle (12) to cover the joint nozzle (12), and the seed pipe (13) is connected with the joint nozzle (12). 11) A second regulating valve (14) is provided, one end of the air outlet pipe (8) is connected with the seed tank body (6), and the other end of the air outlet pipe (8) is connected with a third regulating valve (15) ), the third regulating valve (15) is connected with a flow meter (16), the air outlet pipe (8) is connected with a pressure gauge (17), the exhaust pipe (9) is connected with the air outlet pipe (8) connected, a fourth regulating valve (18) is provided on the emptying pipe (9). 2. the inoculation device of Monascus liquid submerged fermentation production of Monascus red according to claim 1, is characterized in that, described air inlet pipe (7) is connected with the bottom end of described seed tank (6), described The connection between the seed passing pipe (11) and the seed tank (6) and the connection between the air outlet pipe (8) and the seed tank (6) are located on the top of the seed tank (6). . 3. the inoculation device of Monascus liquid submerged fermentation to produce Monascus red according to claim 1, is characterized in that, described inoculation bottle also comprises fixed jacket, and described fixed jacket comprises the first splint (19), The second splint (20) and the locking bolt (21), the first splint (19) is located above the bottle stopper (2), the second splint (20) is sleeved at the bottle mouth, The first clamping plate (19) is provided with two symmetrically distributed first through holes, the second clamping plate (20) is provided with two symmetrically distributed second through holes, and locks are arranged in the second through holes A locking nut, the locking bolt (21) passes through the first through hole and is connected with the locking nut. 4. the inoculation device of Monascus liquid submerged fermentation to produce Monascus red according to claim 1, is characterized in that, described joint mouth (12) comprises fixing part (1201) and connecting part (1202), described fixing Both the part (1201) and the connecting part (1202) are cylindrical, and the diameter of the fixing part (1201) is larger than the diameter of the connecting part (1202), and one end of the connecting part (1202) is connected to the One end of the fixing part (1201) is coaxially fixedly connected, the fixing part (1201) is connected with the end of the seed feeding pipe (11), and the inner wall of the seeding head (5) is provided with a first internal thread, The outer wall of the connecting part (1202) is provided with a first external thread matched with the first internal thread, the inner wall of the joint nozzle cover (13) is provided with a second internal thread, and the fixing part (1201) The outer wall is provided with a second external thread matched with the second internal thread. 5. an inoculation method for producing Monascus red by liquid submerged fermentation of Monascus, is characterized in that, comprises the steps: Step 1: after adding culture medium in the seed tank (6), carry out actual elimination; Step 2: After the actual elimination, sterile air is introduced into the seed tank (6) through the air inlet pipe (7), and the first regulating valve (10) and the third regulating valve (15) are adjusted to make the seed tank (6) Keep a certain positive pressure inside, the positive pressure is ≥ 0.08MPa and the positive pressure is less than 0.1MPa, and the pressure is maintained; Step 3: Sterilize the inoculation bottle, pour the seed liquid of the shaker bottle into the inoculation bottle body (1) using aseptic operation technology in the sterile room, and plug the inoculation bottle body (1) with the bottle stopper (2). Then use a fixed jacket to compress the bottle stopper (2), wrap the connecting tube (3) on the inoculation bottle body (1), and take it to the workshop for inoculation; Step 4: Under the protection of the fire ring, remove the joint nozzle cover (13), quickly remove the protective layer wrapped by the seed head (5), and connect the seed head (5) to the joint nozzle (12) ; After the seed head (5) is connected with the joint mouth (12), fully open the second regulating valve (14), so that the inoculation bottle (1) is communicated with the seed tank (6) and the pressure is consistent; remove the fire ring ; Step 5: During inoculation, control the maximum pressure in the seed tank (6) not to exceed 0.1MPa, slowly open the fourth regulating valve (18), and discharge the gas in the seed tank (6), so that the seed tank ( 6) The pressure in the inoculation bottle body (1) drops, and the seed liquid in the inoculation bottle body (1) is pressed into the seed tank body (6), wherein the seed liquid in the inoculation bottle body (1) is transferred to the seed tank body (6) When the fourth regulating valve (18) is not fully opened, by adjusting the fourth regulating valve (18), the seed liquid is transferred several times until all the seed liquid in the inoculation bottle (1) is transferred to the seed tank (1). 6) in; Step 6: Disconnect the seeding head (5) from the joint nozzle (12), open the second regulating valve (14) halfway, spray a small amount of mycelium from the joint nozzle (12), and clean the joint nozzle (12) , close the second regulating valve (14), cover the joint mouth cover (13) on the joint mouth (12), and complete the inoculation. 6. the inoculation method of Monascus liquid submerged fermentation production of Monascus red according to claim 5, is characterized in that, in step 1, when carrying out actual elimination, by air inlet pipe (7) to seed tank (6) Steam is introduced into it, at this time the first regulating valve (10), the third regulating valve (15) and the fourth regulating valve (18) are opened, when the temperature in the seed tank (6) rises to 121°C, the pressure gauge ( 17) When it is 0.11MPa, fully open the second regulating valve (14), and unscrew the joint nozzle cover (13), the steam is discharged from the joint nozzle (12), and then connect the joint nozzle cover (13) with the joint nozzle (12). ) connection, and control the degree of tightening of the joint nozzle cover (13), so that the steam is obviously discharged from the joint nozzle (12), and then, the temperature in the seed tank (6) is maintained at 121 ~ 124 ℃ for 30 ~ 35min, Finally, the second regulating valve (14) is kept fully open, the joint nozzle cover (13) is tightened, and the fourth regulating valve (18) is closed. 7. the inoculation method of Monascus liquid submerged fermentation to produce Monascus red according to claim 5, is characterized in that, in step 3, when inoculation bottle is sterilized, bottle stopper (2) is tightly plugged inoculation bottle After the bottle mouth of the body (1), wrap the top of the inoculated bottle body (1) with kraft paper, wrap the bottle stopper (2) with kraft paper, then wrap the fixing jacket with kraft paper separately, and wrap the seed head (5) with kraft paper , wrap the connecting tube (3) on the inoculation bottle (1), place the inoculation bottle and the fixed jacket wrapped with kraft paper in a horizontal sterilizer, keep the temperature at 121-124°C, sterilize for 30 minutes, and sterilize After completion, the inoculation bottle is taken out and placed in a sterile room to cool before use. 8. the inoculation method of Monascus liquid submerged fermentation to produce Monascus red according to claim 5, is characterized in that, in step 4, operator wears fire-resistant insulation gloves and takes off joint mouth cover (13), and After the seeding head (5) and the joint nozzle (12) are connected, wet the fire-resistant and heat-insulating gloves and wrap the joint between the seeding head (5) and the joint nozzle (12) to cool them quickly. 9. the inoculation method of Monascus liquid submerged fermentation production of Monascus red according to claim 5, is characterized in that, in step 4, after the seed head (5) is connected with the joint mouth (12), inoculation The outer casing of the bottle body (1) is provided with a transparent protective cover (22), and the protective cover (22) is not higher than the connecting mouth (4). 10. the inoculation method of Monascus liquid submerged fermentation to produce Monascus red according to claim 5, is characterized in that, in step 5, when opening the 4th regulating valve (18), tilt the inoculation bottle (1) , so that the seed liquid in the inoculation bottle (1) is close to the connecting nozzle (4); when the pressure gauge (17) drops to 0.02MPa, immediately close the fourth regulating valve (18), so that the seed tank (6) in the When the pressure rises to 0.08-0.1MPa, in the process of pressure rise, the inoculation bottle (1) is upright, so that the seed liquid leaves the connecting nozzle (4), and the seed liquid in the connecting pipe (3) returns to the inoculation bottle (1) When the pressure rises, slowly open the fourth regulating valve (18), and circulate in this way until all the seed liquid in the inoculation bottle (1) is transferred to the seed tank (6).

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