Vitamin K2 preparation device and vitamin K2 preparation method

Abstract

The invention discloses a preparation device of vitamin K2, and relates to the technical field of vitamin preparation. The preparation device comprises a tank body, a stirring part rotates forwards in the tank body and the tank body rotates backwards and turns over under the cooperation of a connecting mechanism through operation of a rotating mechanism, and the stirring part stretches out and draws back in a reciprocating mode under the action of a linkage part; the stirring part is adjusted to stretch out and draw back through operation of the switching part so that the stirring part can be attached to the inner wall of the tank body; in the moving process of the switching part, the auxiliary mechanism operates so as to introduce disinfection gas into the tank body. The device has the advantages that the whole raw materials are turned over and disturbed while the fermentation raw materials are stirred through forward and reverse rotation, the raw materials are prevented from being stacked and moving in a large range, the mixing effect of the raw materials and the heating uniformity degree are greatly improved, operation of the device can be switched, and the device is convenient to use. The interior of the fermentation device can be efficiently cleaned, the wall scraping effect is achieved, meanwhile, disinfection and sterilization are assisted, and excellent intelligence and practicability are achieved.

Description

Technical Field

[0001] The invention relates to the technical field of vitamin preparation, and in particular to a device for preparing vitamin K2 and a method for preparing vitamin K2. Background Art

[0002] Vitamin K2, also known as menadione, is an important fat-soluble vitamin. According to the length of the isoprene side chain on C-3, it can be expressed as MK-n (n=1-14, usually between 4-13). Among them, MK-7, compared with other forms of vitamin K2, has better bioavailability, stronger lipophilicity and longer half-life, and is the best choice as a source of vitamin K2 for human body. In addition to its significant effects on blood coagulation, prevention of osteoporosis and cardiovascular disease, MK-7 also has functions and health benefits including cancer prevention, inhibition of Parkinson's disease, and help liver function recovery. As a highly active endogenous vitamin in the human body, the safety of vitamin K2 has been authoritatively recognized by the US FDA, the National Drug Administration, the US Institute of Pharmaceutical Research, the European Parliament and the Council of the European Union. Vitamin K2 (MK-7) has been widely used as a dietary supplement and health product.

[0003] In the process of preparing vitamin K2, the fermentation of raw materials is a key link. In the existing fermentation preparation devices, the raw materials are mostly directly put into the fermentation tank for fermentation. Some fermentation devices lack equipment for efficient stirring of the raw materials, resulting in slow fusion of the raw materials, which in turn affects the quality of the finished product of vitamin K2. In addition, some fermentation equipment is not convenient for cleaning and sterilization inside, which may affect the subsequent fermentation quality when used for the second time. Summary of the invention

[0004] The purpose of the present invention is to provide a device for preparing vitamin K2 and a method for preparing vitamin K2, which solve the problems raised in the above-mentioned background technology.

[0005] To achieve the above object, the present invention provides the following technical solution: a device for preparing vitamin K, comprising:

[0006] Tank;

[0007] A feeding part and a discharging part, wherein the feeding part and the discharging part are respectively arranged at the upper and lower parts of the tank body;

[0008] An input part and an output part, wherein the input part and the output part are respectively arranged at the center of the side of the tank body and at the bottom of the tank body;

[0009] A support, the support being arranged below the tank body;

[0010] A rotating mechanism, a connecting mechanism, a stirring component, a linkage component and a switching component, wherein the rotating mechanism, the connecting mechanism, the stirring component, the linkage component and the switching component are all arranged above the support;

[0011] Through the operation of the rotating mechanism, in cooperation with the connecting mechanism, the stirring member is caused to rotate forward in the tank body and the tank body is caused to reverse and flip; under the action of the linkage member, the stirring member is caused to reciprocate and extend;

[0012] The switching component is operated to adjust the extension and contraction of the stirring component so that the stirring component is in contact with the inner wall of the tank body; during the movement of the switching component, the auxiliary mechanism is operated to introduce sterilizing gas into the tank body.

[0013] Optionally, the rotating mechanism includes:

[0014] The mounting shell has a lower surface fixedly connected to the upper surface of the support, a motor 1 is fixedly connected to the inner wall of the mounting shell, a rotating part of the motor 1 is fixedly connected to a rotating rod, and an end of the rotating rod is fixedly connected to a support frame.

[0015] Optionally, the connecting mechanism includes:

[0016] A fixed bevel gear ring, the side surface of the fixed bevel gear ring is fixedly connected to the side surface of the mounting shell, two rotating drums are rotatably connected to the side surface of the support frame, the opposite ends of the two rotating drums are fixedly connected with bevel gears, the opposite ends of the two rotating drums are fixedly connected with pulley 1, a cover plate is rotatably connected to the upper surface of the tank body, the upper surface of the cover plate is fixedly connected to the inner side of the support frame, the center of the cover plate is provided with an opening for the rotating drum to penetrate and be rotatably connected to the same, a connecting shaft is fixedly connected to the bottom center of the rotating drum, the connecting shaft and the opposite ends of the rotating drum are fixedly connected with pulley 2, and the pulley 1 and the pulley 2 are connected by belt transmission.

[0017] Optionally, the stirring component includes:

[0018] A stirring piece, the inner side of which is fixedly connected to the outer side of the rotary drum, a vertical block is slidably connected to the side of the stirring piece, and a scraper is arranged at one end of the vertical block facing the inner wall of the tank body.

[0019] Optionally, the linkage component includes:

[0020] The top end of the driving member is provided with a toothed structure, and the toothed structure is designed so that the driving member can be rotated with the toothed structure to move relative to the toothed structure, and the toothed structure is connected with the toothed structure in a fixed manner.

[0021] Optionally, the switching component includes:

[0022] An electric push rod, wherein the outer shell of the electric push rod is fixedly connected to the upper surface of the cover plate, the telescopic portion of the electric push rod is fixedly connected to a connecting plate, one end of the connecting plate is fixedly connected to the side surface of the sleeve block, the other end of the connecting plate is fixedly connected to a spline shaft for rotation, an inner spline groove for the spline shaft to pass through and be slidably connected to the inner side of the rotating cylinder is provided, and a cam is fixedly connected to the end of the spline shaft.

[0023] Optionally, the auxiliary mechanism includes:

[0024] A pressure plate, a piston rod is fixedly connected to the side of the pressure plate, the end of the piston rod is fixedly connected to the piston head, the outer side of the piston head is slidably connected to a piston cylinder, the outer shell of the piston cylinder is fixedly connected to the inner wall of the mounting shell, the piston cylinder and the opposite side of the pressure plate are commonly and fixedly connected with a spring 2, an input pipe and an output pipe are fixedly connected to the outer surface of the piston cylinder, a one-way valve is arranged inside the input pipe and the output pipe, the end of the output pipe is connected to the input part, and the input part is rotatably connected to the central side of the tank body.

[0025] A method for preparing vitamin K2 comprises the following steps:

[0026] Activation of bacteria: Inoculate Bacillus natto into agar slant medium for activation;

[0027] Bacteria culture: The activated bacteria are inoculated into the activation culture medium and cultured in a shake flask;

[0028] Pretreatment of bacterial strains: pretreat the prepared bacterial cells under the conditions of liquid A;

[0029] The components of liquid A are: amino acid solution and nutrient solution, the amino acid concentration is 0.001-0.1%, and the nutrient solution components are: tryptone 0.2%, yeast powder 0.05%, sodium chloride 0.1%;

[0030] Seed liquid preparation: The pretreated bacteria are inoculated into the seed culture medium and cultured to the logarithmic growth phase, which is the fermentation seed liquid;

[0031] Fermentation in the tank: The prepared seed liquid is aseptically inoculated into the fermentation medium for aerobic fermentation in the tank 1, and liquid B is added during the fermentation process to promote the synthesis of vitamin K2;

[0032] The B solution contains: a physiologically active substance mixture of S-adenosylmethionine, glutathione, coenzyme Q10, and biotin, which is sterile filtered with a 0.22 μm sterile filter when used;

[0033] Fermentation broth centrifugation: centrifuge the fermentation broth obtained from the fermentation to collect the bacterial cells and the supernatant respectively;

[0034] Extraction: The obtained bacteria and supernatant were extracted separately, and the vitamin K2 production was measured by liquid HPLC. The sum of the two vitamin K2 productions was the total fermentation production.

[0035] Compared with the prior art, the present invention has the following beneficial effects:

[0036] 1. The vitamin K2 preparation device operates through a rotating mechanism in cooperation with a connecting mechanism, so that the stirring component rotates forward in the tank body and the tank body is reversed and turned over. The operating mechanism has:

[0037] This method can firstly mix the raw materials in the tank body evenly, and secondly can make the raw materials flow up and down in the tank body. The rotation of the stirring component can make the raw materials for preparing vitamin K mix more evenly to speed up the mixing speed, and avoid the situation where the raw materials are always piled up at the bottom of the tank body and cause poor mixing effect. The raw materials can also be rotated forwardly with the tank body as a whole, and the stirring component can disturb the raw materials to reverse, thereby greatly improving the contact frequency and contact area between the raw materials and the stirring component. With the cooperation of revolution and forward and reverse rotation, the raw materials for fermentation can be fully mixed together to ensure the fermentation effect.

[0038] 2. The vitamin K2 preparation device operates through a connecting mechanism to reciprocate and extend the stirring component under the action of the linkage component. The operating mechanism has:

[0039] This linkage component further improves and optimizes the movement form of the stirring component. Different from the transmission stirring process, this linkage component can make the stirring component extend and contract to a certain extent, which can increase the contact area between the stirring component and the material in the tank body, and the lateral reciprocating telescopic movement can also disturb the raw materials, so that the raw materials can further move irregularly, which can further increase the contact area and contact frequency between the raw materials, thereby ensuring the mixing effect of the raw materials during fermentation and the overall uniformity of heating of the raw materials. In the process of reciprocating extension and contraction of the stirring component, the raw materials can be continuously pushed laterally to improve the fluidity of the raw materials and thus achieve a better stirring effect.

[0040] 3. The vitamin K2 preparation device operates by switching the components to adjust the extension and contraction of the stirring components so that the stirring components are in contact with the inner wall of the tank, and operates the auxiliary mechanism during the movement of the switching components to introduce sterilizing gas into the tank. The operating mechanism has:

[0041] This method allows the vertical blocks to extend and contact the inner wall of the tank before and after fermentation, so as to facilitate cleaning and scraping of the inner wall of the tank. During the fermentation process, the vertical blocks are retracted to avoid the impact of continuous rotation on the inner wall of the tank and to ensure the stirring effect and efficiency, achieving multiple goals at one stroke. Before switching, the fermentation raw materials in the tank can be easily stirred and mixed, and after switching, the materials and impurities on the inner wall of the tank can be easily scraped. During the switching process, the components can also be moved to connect the auxiliary mechanism for operation. Therefore, this switching component is dedicated to the preparation of vitamin K2 and has practical effects before, after and during the switching process.

[0042] When it is necessary to clean the inside of the tank body, the switching component can adjust the stirring component to extend and achieve the effect of rotating and scraping the wall of the tank body. At the same time, the auxiliary mechanism can be operated during the switching process to introduce disinfecting gas into the tank body, so that the tank body can be sterilized and disinfected simultaneously while cleaning the residue inside the tank body. When the tank body is used for fermentation, the auxiliary mechanism will not operate and the stirring component will not extend, so as to avoid affecting the fermentation.

[0043] 4. The preparation method of vitamin K2 can effectively shorten the fermentation cycle and increase the yield of vitamin K2 by pretreating the seed liquid, adding physiologically active substances and optimizing the fermentation process. The natto Bacillus subtilis used in the present invention is a food safety grade strain, the added physiologically active substances can be absorbed and utilized by the human body, and the culture medium formula does not contain heavy metals such as copper. The K2 obtained by fermentation is natural and safe. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1 This is a front view of the vitamin K2 preparation device;

[0045] Figure 2It is a schematic diagram of the structure of the tank body of the present invention;

[0046] Figure 3 It is a schematic diagram of the structure of the installation shell of the present invention;

[0047] Figure 4 It is a schematic diagram of the structure of the cover plate of the present invention;

[0048] Figure 5 It is a schematic diagram of the structure of the vertical block of the present invention;

[0049] Figure 6 It is a schematic diagram of the structure of the rotary drum block of the present invention;

[0050] Figure 7 It is a schematic diagram of the structure of the electric push rod of the present invention;

[0051] Figure 8 It is a schematic diagram of the structure of the set block of the present invention;

[0052] Fig. 9 For the present invention Figure 3 A magnified view of the structure at center A;

[0053] Fig.10 For the present invention Figure 5 A magnified view of the structure at B in the middle;

[0054] Fig.11 A process flow chart of a method for preparing vitamin K2;

[0055] Fig.12 This is a graph showing the effect of bacterial pretreatment on the fermentation yield of vitamin K2 in the present invention;

[0056] Fig.13 This is a graph showing the effect of the feed-batch process for increasing physiologically active substances on the fermentation yield of vitamin K2;

[0057] Fig.14 This is a graph showing the fermentation yield of vitamin K2 from different nitrogen sources in the present invention.

[0058] In the figure: 1, tank body; 2, feeding part; 3, discharging part; 4, input part; 5, output part; 6, support; 7, mounting shell; 8, motor 1; 9, rotating rod; 10, supporting frame; 11, fixed bevel gear ring; 12, rotating drum; 13, bevel gear; 14, pulley 1; 15, cover plate; 16, rotating drum; 17, connecting shaft; 18, pulley 2; 19, stirring member; 20, vertical block; 21, vertical rod; 22, belt transmission mechanism; 23, moving member; 24, insert rod; 25, abutment ring; 26, rod member; 27, hinged plate; 28, connecting block; 29, sleeve block; 30, spring 1; 31, electric push rod; 32, connecting plate; 33, spline shaft; 34, cam; 35, pressure plate; 36, piston rod; 37, piston cylinder; 38, spring 2; 39, protrusion; 40, output pipe. DETAILED DESCRIPTION

[0059] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0060] For example, see Figures 1 to 14 The present embodiment provides a vitamin K2 preparation device, including a tank body 1, a feed part 2, a discharge part 3, an input part 4, an output part 5, a support 6, a rotating mechanism, a connecting mechanism, a stirring component and a linkage component.

[0061] In this embodiment, in actual use, the fermentation medium and the seed liquid are put into the tank body 1 together through the feed part 2. At this time, the rotation mechanism is started, and the rotation mechanism drives the connection mechanism to operate, so that the stirring component can evenly mix and stir the raw materials in the tank body 1, and can make the raw materials turn upside down and disturb, so as to further improve the mixing uniformity of the raw materials and the heating uniformity of the raw materials. Then the linkage component can further change the operation characteristics of the stirring component, and can make the stirring component telescopic during rotation, so that it can play a pushing role on the raw materials to improve the fluidity of the raw materials, so as to greatly improve the mixing and stirring effect of the raw materials, and make the temperature of the raw materials in the tank body 1 37-40°C, the stirring speed 100-800rpm, the ventilation volume 1-2vvm, and control the dissolved oxygen 20%-30%, and ferment for a sufficient time. When the raw materials are mixed in this device, ventilation can be introduced through the input part 4, and the input part 4 can be introduced through the external gas pipe. Since the input part 4 is arranged at the rotation center of the tank body 1, it will not affect the rotation of the tank body 1.

[0062] This embodiment also includes a switching component and an auxiliary mechanism.

[0063] In the present embodiment, in actual use, when the inside of the tank body 1 needs to be cleaned and sterilized before and after fermentation, the switching component is immediately operated, which not only enables the stirring component to extend and rotate and scrape the inner wall of the tank body 1, but also enables the auxiliary mechanism to be connected and operated during the switching process to spray disinfectant gas into the tank body 1, so that cleaning and sterilization are carried out in coordination with each other, so as to improve the cleaning efficiency and effect inside the tank body 1, ensure the cleanliness of the tank body 1, and avoid the breeding of bacteria and affect the secondary use of the device.

[0064] See also Figure 1 , Figure 2 , Figure 3 , Figure 4 , Figure 5 , Figure 6 , Figure 7 and Fig. 9 , the rotating mechanism includes:

[0065] The mounting shell 7 is fixedly connected to the upper surface of the support 6 at the lower surface of the mounting shell 7, and a motor 8 is fixedly connected to the inner wall of the mounting shell 7. The rotating part of the motor 8 is fixedly connected to a rotating rod 9, and the end of the rotating rod 9 is fixedly connected to a support frame 10;

[0066] A fixed bevel gear ring 11 is provided, and the side of the fixed bevel gear ring 11 is fixedly connected to the side of the mounting shell 7. Two rotating drums 12 are rotatably connected to the side of the support frame 10. The opposite ends of the two rotating drums 12 are fixedly connected to bevel gears 13. The opposite ends of the two rotating drums 12 are fixedly connected to pulleys 14. A cover plate 15 is rotatably connected to the upper surface of the tank body 1. The upper surface of the cover plate 15 is fixedly connected to the inner side of the support frame 10. An opening 1 is provided in the center of the cover plate 15 for the rotating drum 16 to penetrate and be rotatably connected to the rotating drum 16. A connecting shaft 17 is fixedly connected to the bottom center of the rotating drum 12. The opposite ends of the connecting shaft 17 and the rotating drum 16 are fixedly connected to pulleys 18. The pulleys 14 and 18 are connected by belt transmission.

[0067] In this embodiment, when the raw materials need to be fermented, when the raw materials are introduced into the interior of the device, the control panel will operate the rotating part of the motor 8, and the rotating part of the motor 8 will drive the rotating rod 9 and the support frame 10 to rotate, so that the tank body 1 can be driven to rotate with the rotating rod 9 as the center, and the raw materials in the tank body 1 can be turned up and down to improve the fluidity of the raw materials. At the same time, the rotating drum 12 and the bevel gear 13 will revolve around the rotating rod 9 as the center, so that under the meshing action with the fixed bevel gear ring 11, the bevel gear 13, the rotating drum 12 and the pulley 14 will rotate. Since the two bevel gears 13 are symmetrically arranged on both sides of the fixed bevel gear ring 11, the upper and lower pulleys 14 rotate in opposite directions. Therefore, under the transmission of the belt and the pulley 2 18, the connecting shaft 17 and the rotating drum 16 will be driven to rotate synchronously in the opposite direction, and the connecting shaft 17 will cause the tank body 1 to rotate forward, and the rotating drum 16 will cause the stirring component to rotate in the reverse direction. This method has the following characteristics:

[0068] In the existing single fermentation equipment without stirring function, the fusion between raw materials is relatively slow when fermenting and preparing vitamins, and it will also affect the actual heating uniformity of the raw materials, which is not conducive to ensuring the quality of the finished vitamins after fermentation. In this method, firstly, the raw materials in the tank body 1 can be evenly mixed, and secondly, the raw materials can be turned upside down and flowed in the tank body 1, so that the raw materials for preparing vitamin K2 can be mixed more evenly with the rotation of the stirring component, so as to speed up the mixing speed and avoid the situation where the raw materials are always piled up at the bottom of the tank body 1 and cause poor mixing effect. In addition, the raw materials can be made to rotate forward along with the tank body 1 as a whole, and the stirring component can disturb the raw materials to reverse, so as to greatly increase the contact frequency and contact area between the raw materials and the stirring component. Under the cooperation of revolution and forward and reverse rotation, the raw materials for fermentation can be fully mixed together to ensure the fermentation effect.

[0069] See also Figure 1 , Figure 2 , Figure 3 , Figure 4 , Figure 5 , Figure 6 , Figure 7 , Fig. 9 and Fig.10 , the rotating mechanism includes:

[0070] The stirring member 19 has its inner side fixedly connected to the outer side of the rotary drum 16 , and a vertical block 20 is slidably connected to the side of the stirring member 19 . A scraper is provided at one end of the vertical block 20 facing the inner wall of the tank body 1 .

[0071] In this embodiment, during actual use, the rotation of the drum 16 will drive the stirring member 19 and the vertical block 20 to rotate, so as to achieve the purpose of stirring and mixing the raw materials in the tank body 1, so that the mixing between the fermentation raw materials is more sufficient and the heating of the raw materials is more uniform, so as to ensure the actual fermentation effect.

[0072] See also Figure 1 , Figure 2 , Figure 3 , Figure 4 , Figure 5 , Figure 6 , Figure 7 , Figure 8 and Fig. 9 , the linkage components include:

[0073] A vertical rod 21, the bottom of which is connected to the upper surface of the cover plate 15 by a fixed axis, the vertical rod 21 is connected to the rotary drum 16 by a belt transmission mechanism 22, a reciprocating screw groove is provided at the upper end of the vertical rod 21, and a moving member 23 is connected to the vertical rod 21 by a reciprocating screw groove thread, a plug rod 24 is fixedly connected to the upper surface of the cover plate 15, a sliding opening is provided on the lower surface of the moving member 23 for the plug rod 24 to pass through and be slidably connected thereto, and the end of the moving member 23 is fixedly connected to The ring 25 and the center of the rotary cylinder 16 are slidably connected with a rod 26, the lower end of the rod 26 is hinged with a hinge plate 27 through a bracket, the end of the hinge plate 27 is hinged with the side of the vertical block 20, the upper end of the rod 26 passes through the rotary cylinder 16 and is connected with a connecting block 28 for fixed axis rotation, the upper end of the connecting block 28 is slidably connected with a sleeve block 29, the inner wall of the sleeve block 29 and the opposite side of the connecting block 28 are fixedly connected with a spring 30, and the side of the rod 26 is fixedly connected with a protrusion 39.

[0074] In this embodiment, in actual use, the rotation of the rod 26 drives the vertical rod 21 to rotate under the transmission of the belt transmission mechanism 22. With the continuous rotation of the vertical rod 21, and the sliding restriction of the moving member 23 by the threaded fit and the insertion rod 24, the moving member 23 and the ring 25 are vertically reciprocating. When the ring 25 moves downward, it squeezes the protrusion 39 and the rod 26 to move downward, and pulls up the spring 1 30. When the ring 25 moves upward, the elastic potential energy of the spring 1 30 is immediately released, so that the rod 26 moves up and resets. Therefore, the rod 26 can move vertically reciprocatingly, and then with the cooperation of the stirring component, the vertical block 20 is reciprocated to extend. In the rotation form of the vertical block 20 and the stirring member 19, the stirring member can continuously extend when rotating to achieve the effect of efficiently stirring the material. This method has the following advantages:

[0075] This linkage component is a further form of the rotating mechanism, so as to further improve and optimize the movement form of the stirring component. Different from the stirring process of the transmission, when the fermentation preparation device stirs the fermentation raw materials, it not only has the stirring effect but also has the effect of flipping and mixing the whole raw materials. When stirring, the stirring component and the tank body 1 itself rotate synchronously in the opposite direction to further improve the stirring. The linkage component can make the stirring component extend and contract to a certain extent without touching the inner wall of the tank body 1, so as to increase the contact area between the stirring component and the material in the tank body 1, and the lateral reciprocating telescopic motion can also disturb the raw materials, so that the raw materials further move irregularly, and the irregular motion can further increase the contact area and contact frequency between the raw materials, so as to ensure the mixing effect of the raw materials during fermentation and the overall uniformity of heating of the raw materials. In the process of reciprocating extension and contraction of the stirring component, the raw materials can be continuously pushed laterally to improve the fluidity of the raw materials, so as to achieve a better stirring effect.

[0076] See also Figure 1 , Figure 3 , Figure 4 , Figure 5 , Figure 6 , Figure 7 , Figure 8 , Fig. 9 and Fig.10 , the switching components include:

[0077] The electric push rod 31 has an outer shell that is fixedly connected to the upper surface of the cover plate 15, and a connecting plate 32 is fixedly connected to the telescopic portion of the electric push rod 31. One end of the connecting plate 32 is fixedly connected to the side surface of the sleeve block 29, and the other end of the connecting plate 32 is rotatably connected to a spline shaft 33. An inner spline groove is provided on the inner side of the rotating drum 12 for the spline shaft 33 to pass through and is slidably connected thereto, and a cam 34 is fixedly connected to the end of the spline shaft 33.

[0078] In this embodiment, before and after fermentation, the control panel automatically controls the telescopic portion of the electric push rod 31 to operate, and the telescopic portion of the electric push rod 31 moves downward and drives the connecting plate 32 and the sleeve block 29 to move downward. As the sleeve block 29 moves downward, it presses and drives the spring 30 and the rod 26 to move downward. Since the linkage component is not rigidly connected to the rod 26, it will not affect the normal downward movement of the rod 26. Fig.10 As shown, under the hinged relationship and the sliding restriction relationship between the stirring member 19 and the vertical block 20, the vertical block 20 moves toward the inner wall of the tank body 1, and finally the scraper block contacts the inner wall of the tank body 1. At the same time, the connecting plate 32 also drives the spline shaft 33 to move downward, and the cam 34 moves downward with the downward movement, and finally the cam 34 faces the pressure plate 35. Therefore, the auxiliary mechanism can be turned on and operated during the switching process. This operating mechanism has the following features:

[0079] During normal fermentation in the tank body 1, the stirring parameter is usually 100-800rpm. When the raw materials are continuously stirred and discharged, some of the raw materials will inevitably adhere to the inner wall of the tank body 1, causing adhesion to the wall, which affects the amount of discharge and the subsequent fermentation. If a scraper is used to always contact the inner wall of the tank body 1, because the stirring component needs to rotate continuously and has a certain speed, when the scraper rotates continuously, although it can avoid adhesion and facilitate cleaning, long-term use will inevitably cause wear of the components or the inner wall, which is not conducive to the service life of the structure and components. Therefore, this invention The vertical block 20 is extended to contact the inner wall of the tank body 1 before and after fermentation to facilitate cleaning and scraping of the inner wall of the tank body 1. During the fermentation process, the vertical block 20 is contracted to avoid the impact of continuous rotation on the inner wall of the tank body 1 and to ensure the stirring effect and efficiency, achieving multiple goals at one stroke. Before switching, the fermentation raw materials in the tank body 1 can be easily stirred and mixed, and after switching, the materials and impurities on the inner wall of the tank body 1 can be easily scraped. During the switching process, the components can be moved to connect the auxiliary mechanisms for operation. Therefore, this switching component is dedicated to the preparation of vitamin K2 and has practical effects before, after and during the switching process.

[0080] For example 2, please refer to Figure 1 , Figure 2 , Figure 3 , Figure 4 , Figure 5 , Figure 6 and Fig. 9 , supporting institutions include:

[0081] The pressure plate 35 has a piston rod 36 fixedly connected to the side of the pressure plate 35, and a piston head fixedly connected to the end of the piston rod 36. The outer side of the piston head is slidably connected to a piston cylinder 37. The outer shell of the piston cylinder 37 is fixedly connected to the inner wall of the mounting shell 7. The piston cylinder 37 and the opposite side of the pressure plate 35 are commonly and fixedly connected with a spring 2 38. The outer surface of the piston cylinder 37 is fixedly connected with an input pipe and an output pipe 40. Both the input pipe and the output pipe 40 are provided with a one-way valve. The end of the output pipe 40 is connected to the input part 4, and the input part 4 is rotatably connected to the central side of the tank body 1.

[0082] In this embodiment, under the action of the switching component, the spline shaft 33 and the cam 34 will move downward, and then the cam 34 will move downward to a position in contact with the pressure plate 35. Therefore, with the self-rotation of the spline shaft 33 and the flipping of the spline shaft 33 driven by the flipping of the tank body 1 itself, the spline shaft 33 will continuously contact and press against the pressure plate 35 during the self-rotation, and under the elastic reset of the spring 2 38, the pressure plate 35, the piston rod 36 and the piston head will reciprocate, so that the pressure in the piston cylinder 37 changes, so that the sterilizing gas can be extracted through the input pipe and input into the tank body 1 through the output pipe 40 from the input part 4. Therefore, after the component switches the form of the stirring component, the auxiliary mechanism can be connected during the switching process, so as to achieve the purpose of introducing sterilizing gas into the tank body 1. This method has the following advantages:

[0083] When it is necessary to clean the inside of the tank body 1, the stirring component can be adjusted to extend under the action of the switching component and achieve the effect of rotating and scraping the wall of the tank body 1. At the same time, the auxiliary mechanism can be operated during the switching process to introduce disinfectant gas into the tank body 1, so that the tank body 1 can be sterilized and disinfected simultaneously when the residue inside the tank body 1 is cleaned. When the tank body 1 is used for fermentation, the auxiliary mechanism will not operate and the stirring component will not extend, thereby avoiding the situation that affects the fermentation. Because in the tank fermentation step in the preparation process of vitamin K2, it is necessary to clean and disinfect the used tank body 1 before and after a single fermentation to avoid impurities and germs in the tank body 1 and affect the fermentation effect. Therefore, this method can quickly clean and disinfect the tank body 1 under the action of the switching component before and after the use of the tank body 1, and can also focus on rotating and scraping the inner wall of the tank body 1, which helps to ensure the effect and efficiency of cleaning and disinfection, so that the disinfectant gas introduced can be steam or ozone gas to achieve the purpose of sterilization and disinfection.

[0084] The present invention also provides a method for preparing vitamin K2, comprising the following steps:

[0085] Step S1: Activation of bacteria: Inoculate Bacillus natto into agar slant medium for activation;

[0086] In step S1 of the present invention, Bacillus natto is deposited in China Center for Type Culture Collection (CCTCC) with the accession number of CCTCCM2020246;

[0087] In step S1 of the present invention, the components of the agar slant culture medium include 1% tryptone, 0.5% yeast extract powder, 1% sodium chloride, 1.5% agar powder, and pH 7.0;

[0088] Step S2: bacterial strain culture: the bacterial strain activated in step S1 is inoculated into the activation culture medium and cultured in a shake flask;

[0089] In step S2 of the present invention, the activation culture medium components include 1% tryptone, 0.5% yeast extract powder, 1% sodium chloride, and pH 7.0;

[0090] Step S3: pretreatment of bacterial strains: pretreatment of the bacterial strains prepared in step S2 under the conditions of liquid A;

[0091] The components of liquid A are: amino acid solution and nutrient solution, the amino acid concentration is 0.001-0.1%, and the nutrient solution components are: tryptone 0.2%, yeast powder 0.05%, sodium chloride 0.1%;

[0092] Step S4: preparing seed solution: inoculating the bacteria pretreated in step S3 into a seed culture medium and culturing them to the logarithmic growth phase, which is the fermentation seed solution;

[0093] The seed culture medium in step S4 of the present invention comprises 1%-5% nitrogen source, 0.5%-5% yeast powder, 0.1%-5% glucose, 1%-2% sodium chloride, and 0.05%-1% dipotassium hydrogen phosphate;

[0094] The culture conditions of step S1, step S2 and step S4 are cultured at a temperature of 30-40°C and 100-300rpm;

[0095] Step S5: tank fermentation: the seed liquid prepared in step S4 is aseptically inoculated into the fermentation medium for aerobic fermentation in tank 1, and liquid B is added during the fermentation process to promote the synthesis of vitamin K2;

[0096] Liquid B contains a mixture of physiologically active substances including S-adenosylmethionine, glutathione, coenzyme Q10, and biotin, which is sterile filtered with a 0.22 μm sterile filter when used.

[0097] In step S5 of the present invention, the fermentation medium components include 1%-20% nitrogen source, 1-10% yeast powder, 1-20% glycerol, 0.1%-5% glucose, 0.05%-1% dipotassium hydrogen phosphate, 0.05%-1% potassium dihydrogen phosphate, and 0.1%-1% magnesium sulfate heptahydrate;

[0098] The fermentation culture conditions in step S5 are to adjust the pH to 6-9 with alkaline solution, culture at a temperature of 30-40° C. and a stirring speed of 100-800 rpm for 48-96 hours, and the alkaline solution is one or more of potassium hydroxide, sodium hydroxide, and ammonia water;

[0099] In the present invention, in step S5, the fermentation process starts to feed the feed medium after 12-48 hours of fermentation, and the feed medium formula is: 10%-30% glycerol, 0.01-0.1% B solution;

[0100] The nitrogen source in the culture medium of the present invention is preferably one or more of soy peptone, soybean meal hydrolyzate, soybean cake powder, and corn flour;

[0101] For each component in solution B:

[0102] S-adenosylmethionine: It has the functions of transmethylation, transsulfuration, and transaminopropylation. It participates in more than 40 biochemical reactions in the body and is closely related to the synthesis of proteins, nucleic acids, neurotransmitters, phospholipids, and vitamins.

[0103] Glutathione: It plays a direct or indirect role in biological functions such as protein and DNA synthesis, material transport, enzyme activity, metabolism and cell protection. The cofactor of many enzyme reactions is also GSH. It is mainly involved in the tricarboxylic acid cycle and sugar metabolism of organisms.

[0104] Coenzyme Q10: Similar in structure to vitamin K, it plays a key role in generating ATP within cells and plays an important role in a series of biological processes such as regulating gene expression, signal transduction, protein transport, and regulating metabolism;

[0105] Biotin: It binds to enzymes and participates in the fixation and carboxylation of carbon dioxide in the body. It is related to important metabolic processes in the body, such as the carboxylation of pyruvate to oxaloacetate, the carboxylation of acetyl-CoA to malonyl-CoA, and other major biochemical reactions in sugar and fat metabolism.

[0106] Step S6: centrifugation of the fermentation broth: centrifuging the fermentation broth obtained in step S5 to collect the bacterial cells and the supernatant respectively;

[0107] Step S7: Extraction: The bacterial cells and supernatant obtained in step S6 are extracted separately, and the vitamin K2 production is measured by liquid HPLC. The sum of the two vitamin K2 productions is the total fermentation production.

[0108] Problems with existing technologies:

[0109] MK-7 can be obtained through direct extraction, fermentation or chemical synthesis. Natto is currently the main raw material for the direct extraction method. Every 100g of natto contains about 800-900μg of vitamin K2, which is a very low content and leads to high production costs. The biologically active MK-7 is an all-trans structure, while the chemical synthesis method will produce a low-activity cis isomer, and there are problems such as limited sources of chemical precursor raw materials, many by-products, low yield, and environmental pollution. The microbial fermentation method can obtain more active natural vitamin K2, but the original wild-type strain screened has problems such as low yield and long fermentation cycle.

[0110] Patent CN107475312B discloses a method for improving cell membrane permeability through the synergistic effect of non-ionic surfactant and ultrasonic irradiation, thereby further effectively increasing the production of vitamin K2. Although the K2 production is greatly improved before and after the treatment, the fermentation liquid needs to be ultrasonically treated every 24 hours and the fermentation time is as long as more than 200 hours. The operation is cumbersome and the fermentation cycle is long. In addition, the ultrasonic treatment of the fermentation liquid is not suitable for industrial production.

[0111] Patent CN110699272A discloses a natto Bacillus subtilis and a method for producing MK-7, wherein a high-yield MK-7 strain is obtained by ultraviolet mutagenesis, and organic or inorganic substances are added to the culture medium to promote the growth of microorganisms and increase the yield of vitamin K2, because the added germination-promoting substances contain heavy metals such as copper, which are potentially harmful to the human body;

[0112] Therefore, it is still a difficult problem to find a method to increase the fermentation yield of MK-7 which is safe, effective, easy to operate, has a short fermentation cycle and is suitable for industrial production to reduce production costs.

[0113] In view of some defects in the prior art, the present invention mainly utilizes the pretreatment of the bacteria to enhance the tolerance of the bacteria, promote the growth of the strain and increase the yield of MK-7;

[0114] In addition, safe, human-usable, physiologically active substances are added to the culture medium and the fermentation process is optimized to promote the synthesis of MK-7, further increase the yield of MK-7, and shorten the fermentation cycle. The strain used in the present invention is Bacillus natto, which does not produce endotoxins and is a food safety grade strain. The strain is suitable for industrial production. The present invention is simple to operate, safe and environmentally friendly, and has a short fermentation cycle, and is suitable for industrial production.

[0115] In summary, the advantages of the present invention over the prior art are as follows: the present invention can effectively shorten the fermentation cycle and increase the yield of vitamin K2, namely MK-7, by pretreating the seed liquid, adding physiologically active substances and optimizing the fermentation process, and provide a new idea and method for industrial large-scale production of K2. In addition, the natto Bacillus subtilis used in the present invention is a food safety grade strain, the added physiologically active substances can be absorbed and utilized by the human body, and the culture medium formula does not contain heavy metals such as copper. The K2 obtained by fermentation is natural and safe. The final yield of vitamin K2 of the present invention is increased by 107%-261%.

[0116] Implementation three, based on the above embodiment:

[0117] Seed liquid pretreatment process:

[0118] First: culture medium preparation;

[0119] LB medium: 1% tryptone, 0.5% yeast extract powder, 1% sodium chloride; sterilize at 115℃ for 30min;

[0120] Seed culture medium: 1% tryptone, 0.5% yeast extract powder, 1% sodium chloride, 1% soy peptone, 1% glycerol, sterilize at 115°C for 30 minutes before use;

[0121] Secondly: pretreatment of bacterial strains;

[0122] The activated strain was inoculated into LB liquid medium, cultured at 37°C, 200 rpm, until the logarithmic phase was about 6-8 hours, 20 ml of the fermentation liquid was drawn, centrifuged at 10000 r / min for 2 minutes, the supernatant was removed, and the precipitate was mixed and washed with 0.9% saline; the centrifugation was repeated once, the supernatant was removed, and the precipitate was suspended in one or a mixed solution of arginine or lysine solution (see Table 1 for details), and treated at 37°C, 200 rpm for 1-5 hours;

[0123] Then: seed solution preparation;

[0124] The pretreated strains were inoculated in a seed culture medium shake flask at pH = 7.0 at 5% inoculation amount, cultured at 37°C, 200 rpm until the logarithmic growth phase (about 6-8 hours) and transferred to the fermentation tank as seed liquid;

[0125] The seed culture medium is 1% soy peptone, 0.5% yeast powder, 1% glucose, 1% sodium chloride, and 0.05% dipotassium hydrogen phosphate;

[0126] After: Fermentation medium;

[0127] 10% soy peptone, 5% yeast powder, 10% glycerol, 1% glucose, 0.05% dipotassium hydrogen phosphate, 0.05% potassium dihydrogen phosphate, 0.1% magnesium sulfate heptahydrate, add purified water to 2L, adjust pH to 7.0 with ammonia water, sterilize at 115℃ for 30min, connect cooling water to cool to 30-40℃ after sterilization;

[0128] And: tank fermentation;

[0129] The seed liquid was inoculated into the fermentation tank with a flame at a 10% inoculation rate, and the fermentation parameters were set at 37-40°C, stirring at 100-800rpm, ventilation at 1-2vvm, and dissolved oxygen at 20%-30%. The fermentation was cultured for 96 hours. After the fermentation liquid was extracted, the vitamin K2 (MK-7) yield was detected. The results were as follows: Fig.12 As shown;

[0130] Should Fig.12 It is the effect of bacterial pretreatment on the fermentation yield of vitamin K2, also known as MK-7.

[0131] Embodiment 4, based on Embodiment 2 and Embodiment 3:

[0132] This example specifically describes the addition of a feed-batch process containing physiologically active substances to promote the synthesis of MK-7, further increase the yield of MK-7 and shorten the fermentation cycle;

[0133] The difference from the second and third embodiments is that:

[0134] Pretreatment of bacterial strains: The activated bacterial strains were inoculated into LB liquid medium, cultured at 37°C, 200rpm, and cultured to the logarithmic phase for about 6-8h, 20ml of fermentation liquid was aspirated, centrifuged at 10000r / min for 2min, the supernatant was removed, and the precipitate was mixed and rinsed with 0.9% saline; the centrifugation was repeated once, the supernatant was removed, and the precipitate was suspended in a mixed solution of 0.020% arginine + 0.020% lysine, and treated at 37°C, 200rpm for 3h;

[0135] Feeding process for adding physiologically active substances: After 15-20 hours of fermentation, when pH and dissolved oxygen rise rapidly, feed medium is started at a rate of 20-40 mL / h until the end of fermentation. The formula of feed medium containing physiologically active substances is as follows:

[0136] Feed medium 1: 30% glycerol, 0.01% S-adenosylmethionine;

[0137] Feed medium 2: 30% glycerol, 0.01% glutathione;

[0138] Feed medium 3: 30% glycerol, 0.01% coenzyme Q10;

[0139] Feed medium 4: 30% glycerol, 0.01% biotin;

[0140] Feed medium 5: 10% glycerol, S-adenosylmethionine, glutathione, coenzyme Q10 and biotin mixed solution at a concentration of 0.01%;

[0141] Feed medium 6: 30% glycerol, S-adenosylmethionine, glutathione, coenzyme Q10 and biotin mixed solution at a concentration of 0.05%;

[0142] Feed medium 7: 10% glycerol, S-adenosylmethionine, glutathione, coenzyme Q10 and biotin mixed solution at a concentration of 0.1%;

[0143] After 48 hours of fermentation, the vitamin K2 (MK-7) production results are as follows Fig.13 As shown;

[0144] Should Fig.13 The effect of the feeding process of increasing physiologically active substances on the fermentation yield of vitamin K2.

[0145] Embodiment 5, based on embodiment 4:

[0146] Seed culture medium: 5% soy peptone, 5% yeast powder, 5% glucose, 2% sodium chloride, 1% dipotassium hydrogen phosphate;

[0147] Fermentation medium: 1% soy peptone, 10% yeast powder, 20% glycerol, 5% glucose, 1% dipotassium hydrogen phosphate, 1% potassium dihydrogen phosphate, 1% magnesium sulfate heptahydrate, add purified water to 2L, adjust pH to 8.0 with potassium hydroxide, sterilize at 115℃ for 30min, connect cooling water and cool to 30-40℃ after sterilization;

[0148] Feed medium: 30% glycerol, 0.025% S-adenosylmethionine, 0.025% biotin;

[0149] After 48 hours of fermentation, the vitamin K2 (MK-7) production was 60.61 mg / L.

[0150] Embodiment 6, based on embodiment 4:

[0151] Seed culture medium: 5% soy peptone, 0.5% yeast powder, 0.1% glucose, 1% sodium chloride, 0.5% dipotassium hydrogen phosphate.

[0152] Fermentation medium: 20% soy peptone, 1% yeast powder, 1% glycerol, 0.1% glucose, 0.05% dipotassium hydrogen phosphate, 0.05% potassium dihydrogen phosphate, 0.1% magnesium sulfate heptahydrate, prepare to 2L with purified water, adjust pH to 9.0 with sodium hydroxide, sterilize at 115℃ for 30min, connect cooling water and cool to 30-40℃ after sterilization.

[0153] Feed medium: 30% glycerol, 0.025% glutathione, 0.025% coenzyme Q10.

[0154] After 48 hours of fermentation, the vitamin K2 (MK-7) production was 48.75 mg / L.

[0155] Embodiment 7, based on embodiment 4:

[0156] Soy peptone in the culture medium was replaced with the nitrogen sources listed in Table 3 ;

[0157] Feed medium: 20% glycerol, 0.025% S-adenosylmethionine, 0.025% biotin.

[0158] After 48 hours of fermentation, the production of vitamin K2 (MK-7) was as follows: Fig.14 shown.

[0159] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A vitamin K2 preparation device, characterized in that: include: Tank body (1); A feeding portion (2) and a discharging portion (3), wherein the feeding portion (2) and the discharging portion (3) are respectively arranged at the upper and lower parts of the tank body (1); An input portion (4) and an output portion (5), wherein the input portion (4) and the output portion (5) are respectively arranged at the center of the side of the tank body (1) and at the bottom of the tank body (1); A support (6), wherein the support (6) is arranged below the tank body (1); A rotating mechanism, a connecting mechanism, a stirring component, a linkage component and a switching component are arranged above the support (6); Through the rotating mechanism, in cooperation with the connecting mechanism, the stirring component is caused to rotate forward in the tank body (1) and the tank body (1) is reversed and turned over, and the stirring component is reciprocated and extended under the action of the linkage component; By operating the switching component, the extension and retraction of the stirring component is adjusted so that the stirring component is in contact with the inner wall of the tank body (1), and the auxiliary mechanism is operated during the movement of the switching component to introduce sterilizing gas into the tank body (1).

2. The vitamin K2 preparation device according to claim 1, characterized in that: The rotating mechanism comprises: A mounting shell (7), wherein the lower surface of the mounting shell (7) is fixedly connected to the upper surface of the support (6), a motor (8) is fixedly connected to the inner wall of the mounting shell (7), a rotating part of the motor (8) is fixedly connected to a rotating rod (9), and the end of the rotating rod (9) is fixedly connected to a support frame (10).

3. The vitamin K2 preparation device according to claim 2, characterized in that: The connecting mechanism comprises: A fixed bevel gear ring (11), the side of the fixed bevel gear ring (11) is fixedly connected to the side of the mounting shell (7), the side of the support frame (10) is connected to two rotating drums (12) in a fixed-axis rotational manner, the opposite ends of the two rotating drums (12) are fixedly connected to bevel gears (13), the opposite ends of the two rotating drums (12) are fixedly connected to pulleys (14), the upper surface of the tank body (1) is connected to a cover plate (15) in a fixed-axis rotational manner, the cover plate The upper surface of the cover plate (15) is fixedly connected to the inner side of the support frame (10), the center of the cover plate (15) is provided with an opening for the rotary drum (16) to penetrate and be connected to the rotary drum (16) for fixed axis rotation, the bottom center of the rotary drum (12) is fixedly connected with a connecting shaft (17), the opposite ends of the connecting shaft (17) and the rotary drum (16) are fixedly connected with a second pulley (18), and the first pulley (14) and the second pulley (18) are connected by a belt transmission.

4. The vitamin K2 preparation device according to claim 3, characterized in that: The stirring member comprises: A stirring member (19), the inner side of which is fixedly connected to the outer side of the rotary drum (16), a vertical block (20) is slidably connected to the side of the stirring member (19), and a scraper is provided at one end of the vertical block (20) facing the inner wall of the tank body (1).

5. The vitamin K2 preparation device according to claim 4, characterized in that: The linkage component comprises: A vertical rod (21), the bottom of the vertical rod (21) is connected to the upper surface of the cover plate (15) by a fixed axis rotation, the vertical rod (21) is connected to the rotary drum (16) by a belt transmission mechanism (22), a reciprocating screw groove is provided at the upper end of the vertical rod (21), a moving member (23) is threadedly connected to the vertical rod (21) through the reciprocating screw groove, an insertion rod (24) is fixedly connected to the upper surface of the cover plate (15), a sliding opening for the insertion rod (24) to pass through and be slidably connected to the moving member (23) is provided at the lower surface of the moving member (23), and a stop ring is fixedly connected to the end of the moving member (23) (25), a rod (26) is slidably connected to the center of the rotary cylinder (16), the lower end of the rod (26) is hinged to a hinge plate (27) through a bracket, the end of the hinge plate (27) is hinged to the side of the vertical block (20), the upper end of the rod (26) passes through the rotary cylinder (16) and is connected to a connecting block (28) for fixed axis rotation, the upper end of the connecting block (28) is slidably connected to a sleeve block (29), the inner wall of the sleeve block (29) and the opposite side of the connecting block (28) are fixedly connected to a spring 1 (30), and a protrusion (39) is fixedly connected to the side of the rod (26).

6. The vitamin K2 preparation device according to claim 5, characterized in that: The switching component comprises: An electric push rod (31), wherein the outer shell of the electric push rod (31) is fixedly connected to the upper surface of the cover plate (15), the telescopic portion of the electric push rod (31) is fixedly connected to a connecting plate (32), one end of the connecting plate (32) is fixedly connected to the side surface of the sleeve block (29), the other end of the connecting plate (32) is fixedly connected to a spline shaft (33) for fixed axis rotation, the inner side of the rotating drum (12) is provided with an inner spline groove for the spline shaft (33) to pass through and to be slidably connected thereto, and the end of the spline shaft (33) is fixedly connected to a cam (34).

7. The vitamin K2 preparation device according to claim 6, characterized in that: The auxiliary mechanism includes: A pressure plate (35), a piston rod (36) is fixedly connected to the side of the pressure plate (35), an end of the piston rod (36) is fixedly connected to a piston head, a piston cylinder (37) is slidably connected to the outer side of the piston head, an outer shell of the piston cylinder (37) is fixedly connected to the inner wall of the mounting shell (7), a spring 2 (38) is fixedly connected to the piston cylinder (37) and the opposite side of the pressure plate (35), an input pipe and an output pipe (40) are fixedly connected to the outer surface of the piston cylinder (37), a one-way valve is provided inside the input pipe and the output pipe (40), an end of the output pipe (40) is connected to the input part (4), and the input part (4) is rotatably connected to the central side of the tank body (1).

8. A method for preparing vitamin K2, characterized in that: The following steps are involved: Step S1: Activation of bacteria: Inoculate Bacillus natto into agar slant medium for activation; Step S2: bacterial strain culture: the bacterial strain activated in step S1 is inoculated into the activation culture medium and cultured in a shake flask; Step S3: pretreatment of bacterial strains: pretreatment of the bacterial strains prepared in step S2 under the conditions of liquid A; The components of liquid A are: amino acid solution and nutrient solution, the amino acid concentration is 0.001-0.1%, and the nutrient solution components are: tryptone 0.2%, yeast powder 0.05%, sodium chloride 0.1%; Step S4: preparing seed solution: inoculating the bacteria pretreated in step S3 into a seed culture medium and culturing them to the logarithmic growth phase, which is the fermentation seed solution; Step S5: fermentation in a tank: the seed liquid prepared in step S4 is aseptically inoculated into the fermentation medium for aerobic fermentation in the tank (1), and liquid B is added during the fermentation process to promote the synthesis of vitamin K2; The B solution contains: a physiologically active substance mixture of S-adenosylmethionine, glutathione, coenzyme Q10, and biotin, which is sterile filtered with a 0.22 μm sterile filter when used; Step S6: centrifugation of the fermentation broth: centrifuging the fermentation broth obtained in step S5 to collect the bacterial cells and the supernatant respectively; Step S7: Extraction: The bacterial cells and supernatant obtained in step S6 are extracted separately, and the vitamin K2 production is measured by liquid HPLC. The sum of the two vitamin K2 productions is the total fermentation production.

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