Production device and production method of ganoderma lucidum fermentation liquor

Abstract

The invention belongs to the field of microbial fermentation, and particularly relates to a production device and a production method of ganoderma lucidum fermentation liquor. The invention provides a production device of ganoderma lucidum fermentation broth, the device comprises a control part, a fermentation part and a liquid storage part, the device has the characteristics of small size and integration, can automatically confirm the highest enzyme activity in-tank time, does not need manual sampling to measure the enzyme activity, can effectively save the complexity and time consumption of manual sampling, and has strong stability. And the ganoderma lucidum fermentation liquor extracted by adopting the device in cooperation with specific fermentation parameters has relatively high laccase activity, and a new thought is provided for large-scale production and intelligent management and control of a fermentation factory.

Description

Technical Field

[0001] The present application belongs to the field of microbial fermentation, and specifically relates to a production device and method for Ganoderma lucidum fermentation liquid. Background Art

[0002] Laccase, a member of the polyphenol oxidase family, has important applications in the food industry, removal of environmental pollutants, water purification, and biomass treatment due to its structural and functional properties. However, the industrial application of laccase still faces many challenges, including low production efficiency, low stability, short lifespan, and high price.

[0003] In recent years, two types of culture have been developed for Ganoderma lucidum: solid-state fermentation (SSF) and submerged fermentation (SmF). Numerous studies have been conducted on process optimization for each type of fermentation. Submerged fermentation is relatively simpler to regulate the physiological production of laccase than solid-state fermentation, and high laccase yields are generally easier to obtain using SmF. Previous research has focused on producing high-yield laccase using submerged fermentation in small laboratory fermenters (less than 50 L), and the resulting enzyme activity has been documented to be very high, reaching 400-4000 U / mL. However, most of the literature is about the research on culture medium formula, fermentation conditions, etc., and a small number of studies are about the fermentation endpoint. The current research on the fermentation endpoint is basically to determine the fermentation endpoint by measuring one or several substances. This method has the following technical defects: 1. Lack of online detection means and methods; 2. The measurement requires sampling time and measurement time. It takes a long time after the measurement results are obtained, and the results will be delayed, which may lead to delays in tank loading and have a certain impact on fermentation efficiency and yield; 3. Because fermentation is generally calculated on a daily basis, operators will have to do tedious work such as turning on steam, sampling, and measurement during night shifts.

[0004] In summary, although traditional technologies have been optimized for the submerged fermentation of the white rot fungus Ganoderma lucidum, the industrial production of Ganoderma lucidum-related laccase still faces problems such as low laccase activity, poor stability, and complex process. Summary of the Invention

[0005] Based on this, an embodiment of the present application provides a production device and a production method for Ganoderma lucidum fermentation liquid.

[0006] On one hand, the present application provides a production device for Ganoderma lucidum fermentation liquid, comprising: a control component, a fermentation component and a liquid storage component;

[0007] The control components include: a PLC control system and an air compressor;

[0008] The fermentation components include a first stirring motor, a fermentation tank, a valve component, a pH detection component and a fermentation liquid delivery pipeline;

[0009] The liquid storage component includes a second stirring motor and a liquid storage tank;

[0010] The PLC control system is connected to the air compressor, the first stirring motor, the valve component and the second stirring motor through lines, respectively, for receiving signals and controlling their operation;

[0011] The fermentation tank is equipped with the first stirring motor;

[0012] The valve component is installed at the bottom of the fermentation tank and is connected to the PLC control system through an air pipe. The valve component is controlled by the PLC control system to open and close for discharging the fermentation liquid;

[0013] The pH detection component is used to monitor the pH value of the fermentation liquid in real time and transmit the signal to the PLC control system;

[0014] One end of the fermentation liquid delivery pipeline is connected to the bottom of the fermentation tank, and the other end is connected to the top of the liquid storage tank, for delivering the fermented liquid to the liquid storage tank;

[0015] The second stirring motor is installed on the top of the liquid storage tank.

[0016] In one embodiment, the liquid storage component further includes a temperature detection component; the temperature detection component is installed on the inner wall of the liquid storage tank, and is used to monitor the temperature of the liquid in the liquid storage tank in real time and transmit the signal to the PLC control system.

[0017] In one embodiment, the liquid storage component further includes a jacket; the jacket is wrapped around the outside of the liquid storage tank;

[0018] In one embodiment, the jacket is provided with an inlet and an outlet, and the temperature of the liquid in the liquid storage tank is controlled by passing a medium.

[0019] In one embodiment, the air compressor is connected to the fermentation tank through a pipeline for supplying gas into the fermentation tank.

[0020] In one embodiment, the stirring shaft of the first stirring motor extends into the fermentation tank, driving the stirring blades to rotate, so that the materials in the fermentation tank are evenly mixed.

[0021] In one embodiment, the stirring shaft of the second stirring motor extends into the liquid storage tank, driving the stirring blades to rotate, so that the liquid in the liquid storage tank remains in a uniform state.

[0022] In one embodiment, the ratio of the tank height to the tank diameter of the fermentation tank body is (2.0-2.5):1.

[0023] On the other hand, the present application provides a method for producing Ganoderma lucidum fermentation liquid, which comprises the steps of using the above-mentioned Ganoderma lucidum fermentation liquid production device for production.

[0024] In one embodiment, the method includes providing Ganoderma lucidum, setting fermentation parameters of a production device for Ganoderma lucidum fermentation liquid, and determining the time to add the liquid to the tank when the pH starts to rise from the lowest point.

[0025] In one embodiment, the process further includes a culturing step before the Ganoderma lucidum fermentation.

[0026] In one embodiment, the culture medium comprises 2-4 parts yeast extract powder, 15-25 parts corn steep liquor, 30-32 parts bran, 8-12 parts tobacco stem powder, 0.8-2 parts MgSO4, 0.8-3 parts KH2PO4, and 0.08-0.12 parts vitamin B1;

[0027] In one embodiment, the incubation temperature is 118° C.-125° C., and the incubation time is 25 min-35 min; and

[0028] In one embodiment, the inoculation amount of Ganoderma lucidum is 5v / v%-15v / v%.

[0029] In one embodiment, the fermentation temperature is 28°C-32°C.

[0030] In one embodiment, the ventilation ratio is (0.5-0.75) V / V·min:1 V / V·min.

[0031] In one embodiment, the stirring blade rotates at a speed of 80 rpm-120 rpm.

[0032] In one embodiment, the fermentation time is 45h-108h.

[0033] In one embodiment, the temperature of the liquid in the liquid storage tank is controlled to be 4°C-11°C.

[0034] This application provides a production device for Ganoderma lucidum fermentation broth, comprising a control unit, a fermentation unit, and a liquid storage unit. This integrated device automatically determines the time to discharge the fermentation broth at the highest enzyme activity, eliminating the need for manual sampling and activity measurement. This significantly reduces the time and effort required for manual sampling and offers excellent stability. Furthermore, the slender structure of the tank body combined with the stirring unit enhances liquid turbulence, promotes oxygen dissolution and uniform nutrient distribution, and prevents localized hypoxia or accumulation of metabolic products. A dual-stage stirring control system is employed: the first stirring motor has adjustable speed (e.g., low speed in the early stages to prevent shear damage, high speed in the later stages to promote mass transfer), adapting to the needs of different fermentation stages. Furthermore, a pH sensor provides real-time monitoring of the fermentation broth's pH to prevent decreased bacterial activity due to metabolic acid / base production. Automatic opening and closing, controlled by a PLC, enables rapid and sterile discharge of the fermentation broth, reducing the risk of contamination. The closed conveying system's fermentation broth delivery pipeline is directly connected to the liquid storage tank, preventing contamination or loss of active ingredients caused by open transfer.

[0035] In summary, the Ganoderma lucidum fermentation liquid extracted using this device in combination with specific fermentation parameters has a high laccase activity, providing new ideas for large-scale production and intelligent management and control in fermentation plants. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] In order to more clearly illustrate the technical solutions in the embodiments of the present application and to more fully understand the present application and its beneficial effects, the following is a brief introduction to the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present application. Those skilled in the art can also derive other drawings based on these drawings without inventive effort.

[0037] Figure 1 A schematic diagram of the components of a production device for Ganoderma lucidum fermentation liquid provided in one embodiment of the present application;

[0038] Figure 2 A schematic diagram of a fermentation component and a liquid storage component provided in one embodiment of the present application;

[0039] Figure 3 The effect of storage temperature on the residual enzyme activity of laccase;

[0040] Figure 4 To verify the pH and enzyme activity of Ganoderma lucidum fermentation broth at different enzyme production levels.

[0041] Description of Reference Numerals

[0042] 10. Production equipment for Ganoderma lucidum fermentation liquid;

[0043] 100. Control components; 101. PLC control system; 102. Air compressor;

[0044] 200, fermentation component; 201, first stirring motor; 202, fermentation tank; 203, valve component; 204, pH detection component; 205, fermentation liquid delivery pipeline;

[0045] 300, liquid storage component; 301, second stirring motor; 302, liquid storage tank; 303, temperature detection component; 304, jacket. DETAILED DESCRIPTION

[0046] Below in conjunction with embodiment and example, the application is described in further detail.Should be understood that these embodiment and example are only used to illustrate the application and are not used to limit the scope of the application, and the purpose of providing these embodiment and example is to make the understanding of the disclosure of the application more thorough and comprehensive.It should also be understood that the application can be implemented in many different forms, is not limited to the embodiment and example described herein, and those skilled in the art can make various changes or modifications without violating the connotation of the application, and the equivalent form obtained also falls within the protection scope of the application.In addition, in the description hereinafter, a large amount of specific details are given in order to provide a more complete understanding of the application, and it should be understood that the application can be implemented without one or more of these details.

[0047] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

[0048] the term

[0049] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0050] The terms "and / or", "or / and", and "and / or" used herein include any one of two or more related listed items, and also include any and all combinations of the related listed items, wherein the arbitrary and all combinations include any combination of two related listed items, any more related listed items, or all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in this application, the technical solution undoubtedly includes technical solutions connected by "logical and" and also undoubtedly includes technical solutions connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, the technical solution of all being connected by "logical OR"), and also includes any and all combinations of A, B, C, and D, that is, the combination of any two or any three of A, B, C, and D, and also includes the four-item combination of A, B, C, and D (that is, the technical solution of all being connected by "logical AND").

[0051] In this application, "plurality", "multiple", "multiple times", "multiples", etc., unless otherwise specified, refer to a quantity greater than or equal to 2. For example, "one or more" means one or more than or equal to two.

[0052] As used herein, "combination thereof", "any combination thereof", "any combination thereof" and the like include all suitable combinations of any two or more of the listed items.

[0053] Herein, the “suitable” mentioned in “suitable combination”, “suitable method”, “any suitable method”, etc. shall be based on the ability to implement the technical solution of this application, solve the technical problems of this application, and achieve the expected technical effects of this application.

[0054] In this application, "further", "further", "particularly" and the like are used for descriptive purposes to indicate differences in content, but should not be understood as limiting the scope of protection of this application.

[0055] In this application, the terms "optionally," "optional," and "optional" mean optional or dispensable, i.e., they refer to either option being selected from two parallel options: "with" or "without." If a technical solution contains multiple "optional" clauses, each "optional" clause is independent unless otherwise specified and there are no contradictions or constraints.

[0056] In this application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0057] In this application, when referring to a numerical interval (i.e., a numerical range), unless otherwise specified, the distribution of the optional values within the numerical interval is considered continuous and includes the two numerical endpoints of the numerical range (i.e., the minimum and maximum values), as well as every numerical value between these two numerical endpoints. Unless otherwise specified, when a numerical interval refers only to integers within the numerical interval, it includes the two numerical endpoints of the numerical range, as well as every integer between the two numerical endpoints. In this document, this is equivalent to directly listing each integer. For example, "t is an integer selected from 1 to 10" means that t is any integer selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. In addition, when multiple ranges are provided to describe a feature or characteristic, these ranges may be combined. In other words, unless otherwise specified, ranges disclosed herein should be understood to include any and all subranges subsumed therein.

[0058] Unless otherwise specified, the temperature parameters in this application allow for both constant temperature treatment and temperature fluctuations within a certain temperature range. It should be understood that the constant temperature treatment allows for temperature fluctuations within the accuracy range of instrument control. Fluctuations within ranges such as ±5°C, ±4°C, ±3°C, ±2°C, and ±1°C are permitted.

[0059] In this application, % (w / w) and wt% both refer to weight percentage, % (v / v) refers to volume percentage, and % (w / v) refers to mass volume percentage.

[0060] All documents mentioned in this application are cited as references in this application, just as each document is cited as a reference individually. Unless they conflict with the invention purpose and / or technical solution of this application, the cited documents involved in this application are cited in their entirety and for all purposes. When cited documents are involved in this application, the definitions of relevant technical features, terms, nouns, phrases, etc. in the cited documents are also cited. When cited documents are involved in this application, the examples and preferred embodiments of the cited relevant technical features may also be incorporated into this application as references, but are limited to the ability to implement this application. It should be understood that when the cited content conflicts with the description in this application, the present application shall prevail or be adaptively amended according to the description in this application.

[0061] Previous research has also explored industrial production. Liu et al. (2016) achieved a laccase yield of up to 80 U / mL using a 5-ton high-density culture. While this yield was significantly lower than the 240 U / mL achieved in a 5-L small-scale fermenter under the same conditions, the fermenter was still too small to be suitable for industrial production. While researchers have optimized submerged fermentation of the white-rot fungus Ganoderma lucidum, industrial production of laccase still faces numerous challenges, including low efficiency, low stability, and high cost (Khatami et al., 2022).

[0062] Based on this, the purpose of this application is to provide a method and device for fully automatic production of Ganoderma lucidum fermentation liquid, which can maximize fermentation efficiency and enzyme production and optimize personnel operation through online judgment.

[0063] like Figure 1 The present application provides a production device 10 for a Ganoderma lucidum fermentation liquid, comprising: a control component 100, a fermentation component 200, and a liquid storage component 300;

[0064] The control component 100 includes: a PLC control system 101 and an air compressor 102;

[0065] The fermentation component 200 includes a first stirring motor 201, a fermentation tank 202, a valve component 203, a pH detection component 204 and a fermentation liquid delivery pipeline 205;

[0066] The liquid storage component 300 includes a second stirring motor 301 and a liquid storage tank 302;

[0067] The PLC control system 101 is connected to the air compressor 102, the first stirring motor 201, the valve component 203 and the second stirring motor 301 through lines, respectively, for receiving signals and controlling their operation;

[0068] The air compressor 102 is connected to the fermentation tank 202 via a pipeline, and is used to pass gas into the fermentation tank 202;

[0069] The first stirring motor 201 is installed on the top of the fermentation tank 202. The stirring shaft of the first stirring motor 201 extends into the fermentation tank 202, driving the stirring blade to rotate, so that the materials in the fermentation tank 202 are evenly mixed;

[0070] The valve component 203 is installed at the bottom of the fermentation tank 202 and is connected to the PLC control system 101 through an air pipe. The valve component 203 is controlled by the PLC control system 101 to open and close the valve component for discharging the fermentation liquid.

[0071] The pH detection component 204 is used to monitor the pH value of the fermentation liquid in real time and transmit the signal to the PLC control system 101;

[0072] One end of the fermentation liquid delivery pipe 205 is connected to the bottom of the fermentation tank 202, and the other end is connected to the top of the liquid storage tank 302, for delivering the fermented liquid to the liquid storage tank 302;

[0073] The second stirring motor 301 is installed on the top of the liquid storage tank 302. The stirring shaft of the second stirring motor 301 extends into the liquid storage tank 302 to drive the stirring blades to rotate, so that the liquid in the liquid storage tank 302 remains in a uniform state.

[0074] In one embodiment, the liquid storage component 300 further includes a temperature detection component 303 ; the temperature detection component 303 is installed on the inner wall of the liquid storage tank 302 , and is used to monitor the temperature of the liquid in the liquid storage tank 302 in real time and transmit the signal to the PLC control system 101 .

[0075] In one embodiment, the liquid storage component 300 further includes a jacket 304 ; the jacket 304 is wrapped around the outside of the liquid storage tank 302 .

[0076] In one embodiment, the valve component 203 includes a pneumatic electromagnetic tank bottom valve.

[0077] In one embodiment, the jacket 304 is provided with an inlet and an outlet, and the temperature of the liquid in the liquid storage tank 302 is controlled by introducing a heating or cooling medium.

[0078] In one embodiment, the fermentation tank 202 has a tank height to tank diameter ratio of (2.0-2.5):1. For example, the tank height to tank diameter ratio is 2:1, 2.1:1, 2.2:1, 2.3:1, 2.4:1, or 2.5:1, and any values therebetween.

[0079] Another aspect of the present application provides a method for producing a Ganoderma lucidum fermentation liquid, comprising the steps of using the above-mentioned Ganoderma lucidum fermentation liquid production device 10. The method can automatically determine the time for tanking when the enzyme activity reaches the highest level and automatically tank-store the liquid.

[0080] In one embodiment, the method includes providing Ganoderma lucidum, setting fermentation parameters of a production device 10 for Ganoderma lucidum fermentation liquid, and determining the time to add the liquid to the tank when the pH value starts to rise from the lowest point.

[0081] The specific control procedures for determining the timing of tank placement by pH are as follows:

[0082] def get_current_ph():

[0083] #Sensors should be connected here to obtain real-time data

[0084] # Function to open the tank bottom valve

[0085] def open_valve():

[0086] print("The tank bottom valve is open")

[0087] #Function to pressurize fermentation tank 202

[0088] def apply_pressure(pressure):

[0089] if pressure >= 0.1 and pressure <= 0.3:

[0090] print(f"The pressure in fermentation tank 202 has been increased to {pressure:.2f}MPa")

[0091] else:

[0092] print("Pressure value out of range, operation not performed")

[0093] During the fermentation process, the pH value is monitored to confirm that the time when the pH starts to rise from the lowest point is the most suitable time for Ganoderma lucidum to ferment and produce laccase. The control system controls the pneumatic valve at the bottom of the fermentation tank, controls the valve to open, and sets the air pressure in the tank to 0.1 MPa ~ 0.2 MPa, driving the fermentation liquid to a low-temperature temporary storage tank. Under low-temperature conditions, the fermentation liquid can be temporarily stored for 8 hours and retain more than 90% of the laccase activity.

[0094] This method automatically confirms the time for removing the product from the tank when the highest enzyme activity is reached. There is no need for manual sampling to measure enzyme activity, which completely saves the tedious and time-consuming manual sampling. It can quickly and swiftly confirm the time for removing the product from the tank, greatly improving fermentation efficiency and saving water, electricity and gas consumption. It also realizes automatic storage in the tank, providing new ideas for large-scale production and intelligent management and control in fermentation plants.

[0095] In one embodiment, the process further includes a culturing step before the Ganoderma lucidum fermentation.

[0096] In one embodiment, the culture medium comprises 2-4 parts yeast extract, 15-25 parts corn steep liquor, 30-32 parts bran, 8-12 parts tobacco stem powder, 0.8-1.2 parts MgSO4, 0.8-1.3 parts KH2PO4, and 0.08-0.12 parts vitamin B1; for example, 2 parts, 3 parts, or 4 parts of yeast extract, and any values therebetween; and 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 parts of corn steep liquor, and any values therebetween.

[0097] 30 parts, 31 parts or 32 parts of bran and any values therebetween; 8 parts, 9 parts, 10 parts, 11 parts or 12 parts of tobacco stem powder and any values therebetween;

[0098] For example, MgSO4 0.8 parts, 0.9 parts, 1.0 parts, 1.1 parts or 1.2 parts and any values therebetween; for example, vitamin B1 0.08 parts, 0.09 parts, 0.10 parts, 0.11 parts or 0.12 parts and any values therebetween.

[0099] In one embodiment, the culture conditions include a culture temperature of 118°C to 125°C for 25 min to 35 min, such as 118°C, 119°C, 120°C, 121°C, 122°C, 123°C, 124°C, or 125°C, and any values therebetween.

[0100] For example, the time is 25 min, 26 min, 27 min, 28 min, 29 min, 30 min, 31 min, 32 min, 33 min, 34 min or 35 min, and any value in between.

[0101] In one embodiment, the culture conditions include a Ganoderma lucidum inoculation rate of 5%-15%.

[0102] In one embodiment, the fermentation parameters include a fermentation temperature of 28°C-32°C; 28°C, 29°C, 30°C, 31°C or 32°C and any values therebetween are listed.

[0103] In one embodiment, the fermentation parameters include a ventilation ratio of (0.5-0.75) V / V·min:1 V / V·min; for example, the ventilation ratio is 0.5:1, 0.51:1, 0.52:1, 0.53:1, 0.54:1, 0.55:1, 0.56:1, 0.57:1, 0.58:1, 0.59:1, 0.6:1, 0.61:1, 0.62:1, 0.63:1, 0.64:1, 0.65:1, 0.66:1, 0.67:1, 0.68:1, 0.69:1, 0.7:1, 0.71:1, 0.72:1, 0.73:1, 0.74:1 or 0.75:1 and any value therebetween.

[0104] In one embodiment, the fermentation parameters include a stirring blade rotation speed of 80 rpm-120 rpm; for example, the rotation speed is 80 rpm, 85 rpm, 90 rpm, 95 rpm, 100 rpm, 105 rpm, 110 rpm, 115 rpm or 120 rpm and any value therebetween.

[0105] In one embodiment, the fermentation parameters include a fermentation time of 45h-108h; for example, the fermentation time is 45h, 50h, 55h, 60h, 65h, 70h, 75h, 80h, 85h, 90h, 95h, 100h, 105h or 108h and any value therebetween.

[0106] In one embodiment, the fermentation parameters include controlling the temperature of the liquid in the liquid storage tank 302 to be 4°C-11°C, for example, 4°C, 5°C, 6°C, 7°C, 8°C, 9°C, 10°C, or 11°C.

[0107] The method comprises a fermentation tank, a pH detection component, a temporary storage tank and a control system. During the fermentation process, the inoculation amount is set to 5-15%, the fermentation temperature is set to 28-32°C, the ventilation ratio is set to 0.5-0.75, and the rotation speed is set to 100 rpm. At this time, the maximum laccase activity of the fermentation can reach 214185.2 U / L.

[0108] The embodiments of the present application will be described in detail below with reference to the examples. It should be understood that these examples are intended to illustrate the present application only and are not intended to limit the scope of the present application. The experimental methods for which specific conditions are not specified in the following examples are preferably referred to the guidance provided in the present application, and can also be based on the experimental manuals or conventional conditions in this area, or according to the conditions recommended by the manufacturer, or with reference to experimental methods known in the art.

[0109] In the following specific examples, the measured parameters of raw material components may have slight deviations within the range of weighing accuracy unless otherwise specified. For temperature and time parameters, acceptable deviations caused by instrument testing accuracy or operational accuracy are allowed.

[0110] It should be understood that in the various embodiments of the present application, the size of the serial numbers of the above-mentioned processes does not mean the order of execution. The execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

[0111] Example 1

[0112] like Figure 2 The embodiment shown provides a production device 10 for a Ganoderma lucidum fermentation liquid, comprising: a control component 100 , a fermentation component 200 and a liquid storage component 300 .

[0113] 1. Control Component 100

[0114] The control component 100 includes a PLC control system and an air compressor 102; the PLC control system 101 is connected to the air compressor 102, the first stirring motor 201, the pneumatic electromagnetic tank bottom valve 203 and the second stirring motor 301 through lines, for receiving signals and controlling their operation.

[0115] The air compressor 102 is connected to the fermentation tank 202 through a pipeline and is used to supply gas into the fermentation tank 202.

[0116] 2. Fermentation Components 200

[0117] The fermentation component 200 includes a first stirring motor 201 , a fermentation tank 202 , a valve component 203 , a pH detection component 204 , and a fermentation liquid delivery pipeline 205 .

[0118] A first stirring motor 201 is installed on the top of the fermentation tank 202. The stirring shaft of the first stirring motor 201 extends into the fermentation tank 202, driving the stirring blades to rotate, so that the materials in the fermentation tank 202 are evenly mixed. The ratio of the tank height to the tank diameter of the fermentation tank 202 is 2.0:1.

[0119] The pneumatic electromagnetic tank bottom valve is installed at the bottom of the fermentation tank 202 and is connected to the PLC control system 101 through an air pipe. The PLC control system 101 controls its opening and closing for discharging the fermentation liquid.

[0120] The pH detection component 204 is used to monitor the pH value of the fermentation liquid in real time and transmit the signal to the PLC control system 101.

[0121] One end of the fermentation liquid delivery pipeline 205 is connected to the bottom of the fermentation tank 202 , and the other end is connected to the top of the liquid storage tank 302 , so as to deliver the fermented liquid to the liquid storage tank 302 .

[0122] 3. Liquid storage component 300

[0123] The liquid storage component 300 includes a second stirring motor, a liquid storage tank 302, a temperature detection component 303 and a jacket 304. The temperature detection component 303 is installed on the inner wall of the liquid storage tank 302 and is used to monitor the temperature of the liquid in the liquid storage tank 302 in real time and transmit the signal to the PLC control system 101.

[0124] The jacket is wrapped around the outside of the liquid storage tank 302 and is provided with an inlet and an outlet. The temperature of the liquid in the liquid storage tank 302 is controlled by passing a heating or cooling medium.

[0125] A second stirring motor 301 is installed on the top of the liquid storage tank 302. The stirring shaft of the second stirring motor 301 extends into the liquid storage tank 302 to drive the stirring blades to rotate, so that the liquid in the liquid storage tank 302 remains in a uniform state.

[0126] Example 2

[0127] This embodiment provides a method for producing a Ganoderma lucidum fermentation liquid, which is carried out using the Ganoderma lucidum fermentation tank 202 of Example 1 and includes the following steps:

[0128] 1. Cultivate Ganoderma lucidum

[0129] Culture medium formula: yeast extract powder 3%, corn steep liquor 20.0%, bran 31.6%, tobacco stem powder 10.0%, MgSO4·7H2O 1.0%, KH2PO4 1.1%, vitamin B1 0.1%, sterilization at 121℃ for 30 min, Ganoderma lucidum inoculation amount 10%.

[0130] 2. Set the fermentation parameters of the production device of Ganoderma lucidum fermentation liquid

[0131] The fermentation tank 202 is set with a fermentation temperature of 30°C, a ventilation ratio of 0.60, a rotation speed of 100 rpm, and a culture time of 60 hours. The specific culture time is determined by the control system, and the time to unload the tank is determined when the pH starts to rise from the lowest point. This process is determined and automatically operated by the system.

[0132] The specific control procedures are as follows:

[0133] def get_current_ph():

[0134] #Sensors should be connected here to obtain real-time data

[0135] # Function to open the tank bottom valve

[0136] def open_valve():

[0137] print("The tank bottom valve is open")

[0138] #Function to pressurize fermentation tank 202

[0139] def apply_pressure(pressure):

[0140] if pressure >= 0.1 and pressure <= 0.3:

[0141] print(f"The pressure in fermentation tank 202 has been increased to {pressure:.2f}MPa")

[0142] else:

[0143] print("Pressure value out of range, operation not performed")

[0144] After automatic operation, the tank can be removed when the pH drops to the lowest point or increases by 0.03~0.05.

[0145] 3. Storage in tanks

[0146] The bottom valve of fermentation tank 202 is opened, and the pressure inside the tank rises to 0.2 MPa. The pressure drives the fermentation liquid through the pipeline to the temporary storage tank. The temporary storage tank has a jacket temperature control function, which can control the liquid temperature in the tank to 8°C. Under this condition, the fermentation liquid can be stored for 8 hours and retain more than 90% of the laccase activity.

[0147] The detection steps of laccase activity are as follows: add 2 mL of sodium hydrogen phosphate-citrate buffer with a pH of 3 into a 5 mL EP tube, then add 0.5 mL of appropriately diluted fermentation broth supernatant, then add 0.5 mL of 1 mmol / L ABTS solution, and quickly place in a constant temperature water bath at 45°C for 5 minutes. After the reaction is completed, quickly measure the OD 420 .

[0148] Enzyme activity calculation formula: Laccase (U / L) =

[0149] Where, ε = 3.6 × 10 4 , L / (mol / cm); Vtotal-total volume of reaction system (mL); Venzyme-volume of added fermentation broth (mL); Δt-reaction time (min).

[0150] The dilution factor is determined according to the enzyme activity to ensure that the absorbance is between 0.2 and 0.8.

[0151] Example 3: Effect of storage temperature on residual laccase activity

[0152] In this example, the residual enzyme activity was investigated by adjusting the storage time. Figure 3 As shown, Figure 3 The results show the effect of temperature on the preservation rate.

[0153] At 4°C, the preservation rate decreases most slowly over time, indicating that low temperatures significantly slow down the degradation of the indicator. For example, a 70% preservation rate may require a longer time (e.g., over 80 hours). At 18°C, the preservation rate decreases most rapidly, indicating that high temperatures accelerate quality loss. The preservation rate may drop to 70% in a relatively short period of time (e.g., 30 hours). At 11°C, the temperature falls between the two, reflecting an intermediate temperature effect.

[0154] Example 4: Verification of pH and enzyme activity of Ganoderma lucidum fermentation broth at different enzyme production levels

[0155] This example investigates the pH and enzyme activity of Ganoderma lucidum fermentation broth at different enzyme production levels. Figure 4 shown.

[0156] Depend on Figure 4 The results show that the initial pH of different batches of fermentation broth is different, which is caused by the amount of condensed water during the sterilization process and the error of the calibration pH. In addition, the higher the enzyme activity, the longer the fermentation time corresponding to the lowest pH point. This may be because the pH stable stage is the growth and reproduction stage of Ganoderma lucidum. This period is long, indicating that Ganoderma lucidum grows well. The pH begins to decrease when Ganoderma lucidum rapidly produces laccase. This may be because Ganoderma lucidum consumes carbon and nitrogen sources, and the bacteria metabolize and utilize H + When the pH in the culture medium begins to rise, the enzyme activity is basically stable and even shows a downward trend. This shows that after the pH begins to rise, the Ganoderma lucidum no longer produces laccase. The corresponding decrease in enzyme activity is caused by the instability of laccase to storage temperature and time.

[0157] Therefore, the corresponding laccase has the highest enzyme activity from the lowest pH point to the beginning of the rise, which is suitable for tanking.

[0158] The embodiments described above only express several implementation methods of the present application, which are convenient for understanding the technical solutions of the present application in a specific and detailed manner, but they cannot be understood as limiting the scope of protection of the patent application. It should be pointed out that for ordinary technicians in this field, without departing from the concept of the present application, several variations and improvements can be made, which all fall within the scope of protection of the present application. In addition, it should be understood that after reading the above-mentioned teaching content of the present application, those skilled in the art can make various changes or modifications to the present application, and the equivalent forms obtained also fall within the scope of protection of the present application. It should also be understood that the technical solutions obtained by those skilled in the art through logical analysis, reasoning or limited experiments on the basis of the technical solutions provided in the present application are all within the scope of protection of the claims attached to the present application. Therefore, the scope of protection of the patent application of this application shall be based on the content of the attached claims, and the description can be used to interpret the content of the claims.

Claims

1. A production device for Ganoderma lucidum fermentation liquid, characterized in that: include: Control components, fermentation components and liquid storage components; The control components include: a PLC control system and an air compressor; The fermentation components include a first stirring motor, a fermentation tank, a valve component, a pH detection component and a fermentation liquid delivery pipeline; The liquid storage component includes a second stirring motor and a liquid storage tank; The PLC control system is connected to the air compressor, the first stirring motor, the valve component and the second stirring motor through lines, respectively, for receiving signals and controlling their operation; The fermentation tank is equipped with the first stirring motor; The valve component is installed at the bottom of the fermentation tank and is connected to the PLC control system through an air pipe. The valve component is controlled by the PLC control system to open and close for discharging the fermentation liquid; The pH detection component is used to monitor the pH value of the fermentation liquid in real time and transmit the signal to the PLC control system; One end of the fermentation liquid delivery pipeline is connected to the bottom of the fermentation tank, and the other end is connected to the top of the liquid storage tank, for delivering the fermented liquid to the liquid storage tank; The second stirring motor is installed on the top of the liquid storage tank.

2. The production device of Ganoderma lucidum fermentation liquid according to claim 1, characterized in that: The liquid storage component also includes a temperature detection component; the temperature detection component is installed on the inner wall of the liquid storage tank, and is used to monitor the temperature of the liquid in the liquid storage tank in real time and transmit the signal to the PLC control system.

3. The production device of Ganoderma lucidum fermentation liquid according to claim 1, characterized in that: The liquid storage component further includes a jacket; the jacket is wrapped around the outside of the liquid storage tank; Optionally, the jacket is provided with an inlet and an outlet, and the temperature of the liquid in the liquid storage tank is controlled by passing a medium.

4. The production device of Ganoderma lucidum fermentation liquid according to claim 1, characterized in that: The air compressor is connected to the fermentation tank through a pipeline and is used to introduce gas into the fermentation tank; Optionally, a stirring shaft of the first stirring motor extends into the fermentation tank to drive the stirring blade to rotate, so that the materials in the fermentation tank are evenly mixed; Optionally, the stirring shaft of the second stirring motor extends into the liquid storage tank to drive the stirring blades to rotate, so that the liquid in the liquid storage tank remains in a uniform state.

5. The production device of Ganoderma lucidum fermentation liquid according to claim 1, characterized in that: The ratio of the tank height to the tank diameter of the fermentation tank body is (2.0-2.5):

1.

6. A method for producing a Ganoderma lucidum fermentation liquid, characterized in that: The production method comprises the steps of using the production device for Ganoderma lucidum fermentation liquid according to any one of claims 1 to 5 to carry out production.

7. The method for producing Ganoderma lucidum fermentation liquid according to claim 6, characterized in that: include: Provide Ganoderma lucidum, set the fermentation parameters of the production equipment of Ganoderma lucidum fermentation liquid, and determine the time to add the liquid to the tank when the pH starts to rise from the lowest point.

8. The method for producing Ganoderma lucidum fermentation liquid according to claim 7, characterized in that: The fermentation of Ganoderma lucidum also includes a cultivation step.

9. The method for producing Ganoderma lucidum fermentation liquid according to claim 8, characterized in that: Cultivate one or more of the following conditions: (1) The culture medium includes 2-4 parts yeast extract powder, 15-25 parts corn steep liquor, 30-32 parts bran, 8-12 parts tobacco stem powder, 0.8-1.2 parts MgSO4, 0.8-1.3 parts KH2PO4, and 0.08-0.12 parts vitamin B1; (2) The incubation temperature is 118°C-125°C and the incubation time is 25 min-35 min; and (3) The inoculation amount of Ganoderma lucidum is 5v / v%-15 v / v%.

10. The method for producing a Ganoderma lucidum fermentation liquid according to any one of claims 6 to 9, characterized in that: Fermentation parameters meet one or more of the following conditions: (1) Fermentation temperature is 28℃-32℃; (2) Ventilation ratio is (0.5-0.75) V / V·min:1 V / V·min; (3) The stirring blade rotates at a speed of 80 rpm-120 rpm; (4) Fermentation time is 45h-108h; and (5) Control the temperature of the liquid in the storage tank to 4℃-11℃.

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