CN117887776A - Fermentation method of vitamin k2 - Google Patents
Fermentation method of vitamin k2 Download PDFInfo
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- CN117887776A CN117887776A CN202311701556.9A CN202311701556A CN117887776A CN 117887776 A CN117887776 A CN 117887776A CN 202311701556 A CN202311701556 A CN 202311701556A CN 117887776 A CN117887776 A CN 117887776A
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- 230000004151 fermentation Effects 0.000 title claims abstract description 76
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- 238000000034 method Methods 0.000 title claims abstract description 26
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- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 claims description 7
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The application relates to the technical field of microbial fermentation, and provides a fermentation method of vitamin K2, which comprises the following steps of firstly selecting a proper strain, namely bacillus subtilis, secondly preparing a fermentation culture medium, wherein a carbon source selects 20-25 g/L of glucose, a nitrogen source selects 10-12 g/L of peptone, and a proper amount of trace elements and vitamins are added for promoting the growth of the bacillus subtilis and the production of vitamin K2, thirdly, performing fermentation, wherein the temperature is maintained between 35 ℃ and 40 ℃, the pH value is maintained between 6.8 and 7.2, the humidity is controlled between 65% and 75%, and the oxygen supply is controlled between 0.2vvm and 0.4vvm through the gas flow rate. The bacillus subtilis is combined with a culture medium containing glucose, peptone, trace elements and vitamins, so that the growth of bacillus natto and the production of vitamin K2 are promoted, the production capacity of the vitamin K2 is enhanced, and the yield and the production efficiency of the vitamin K2 can be further improved.
Description
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to a fermentation method of vitamin k 2.
Background
Vitamin K2 exerts its biological activity in humans by activating some key proteins, such as bone morphogenic proteins and clotting factors, vitamin K2 is mainly derived from microorganisms, also known as menaquinones, which are known as "platinum vitamins" in food in very small amounts, which are critical for the normal functioning of skeletal health, cardiovascular system and nervous system, and as the awareness of health increases, the demand for vitamin K2 increases gradually,
at present, the preparation method of vitamin K2 mainly comprises a chemical synthesis method and a microbial fermentation method, and the production of vitamin K2 by microbial fermentation mainly comprises the use of bacillus natto, flavobacterium, lactobacillus and the like, but the whole process is more complicated, and the methods have the problems of more waste discharge, high energy consumption, low product purity, lower production efficiency and the like.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a fermentation method of vitamin k2, which solves the problems of more waste emission, high energy consumption, low product purity and lower production efficiency of the whole process.
In order to achieve the purpose, the invention is realized by the following technical scheme: a fermentation method of vitamin k2, comprising the steps of;
step one, selecting a proper strain, wherein the strain is bacillus subtilis;
step two, preparing a fermentation culture medium, wherein a carbon source selects 20 to 25g/L of glucose, a nitrogen source selects 10 to 12g/L of peptone, and a proper amount of trace elements and vitamins are added for promoting the growth of bacillus natto and the production of vitamin K2;
step three, fermentation is carried out, wherein the temperature is maintained between 35 ℃ and 40 ℃, the pH value is maintained between 6.8 and 7.2, the humidity is controlled between 65% and 75%, the oxygen supply is controlled between 0.2 and 0.4vvm through the gas flow rate, and the optimal growth and enzyme producing environment is ensured through a real-time monitoring technology;
step four, adding proper auxiliary nutrients such as coenzyme Q10 and vitamin D in the fermentation process, wherein the auxiliary nutrients are used for promoting the growth of the strain and the synthesis of vitamin K2;
step five, extracting by using dimethylbenzene as a solvent, wherein the soaking time is controlled to be 18 to 30 hours, and the extracting time is controlled to be 3 to 5 hours;
step six, using column chromatography technology, carrying out high-efficiency separation by using a C18 reverse phase column with the diameter of 10mm and the length of 300mm, gradually increasing the solvent gradient from 20% acetonitrile to 80% acetonitrile, and enabling the flow rate to be 1.0mL/min so as to remove impurities and improve the purification effect;
step seven, using 0.1 to 0.3 percent of natural flavonoid compounds, 0.1 to 0.2 percent of vitamin E as an antioxidant and 0.05 to 0.1 percent of benzoic acid as a preservative for enhancing the stability of the extracting solution;
and step eight, carrying out comprehensive quality detection on the vitamin K2 by using a high-resolution analysis technology.
Preferably, in the first step, the bacillus subtilis is a bacterium with stronger heat resistance, has good fermentation capacity, is adaptive to different environmental conditions, and provides a certain stability for the production of vitamin K2.
Preferably, in the second step, the trace elements are 0.3 to 0.5 g of calcium, 0.01 to 0.02 g of zinc and 0.1 to 0.2 g of magnesium, and the vitamin is vitamin D, so that the utilization efficiency of the strain on nutrient components in the culture medium is improved, and the production capacity of vitamin K2 is enhanced.
Preferably, in the third step, the reaction engineering principle is used during fermentation, and each parameter in the fermentation process is precisely controlled through mathematical modeling and optimization algorithm, so that the yield of vitamin K2 is improved to the maximum extent.
Preferably, in the third step, renewable energy sources such as solar energy and wind energy are used in the fermentation process, so that the dependence on the traditional energy sources is reduced, and the environmental impact is reduced.
Preferably, in the eighth step, the high-resolution analysis technology includes high performance liquid chromatography or mass spectrometry, which can perform comprehensive quality detection on the vitamin K2, so as to ensure that the vitamin K2 meets national and international relevant regulations and standards.
Preferably, in the second step, the biodegradable material is used for preparing the fermentation medium and the production equipment, so that the dependence of the whole production process on plastics and other non-degradable materials is reduced.
Preferably, in the third step, a water circulation system is used, so that the water consumption is reduced to the greatest extent, and the water is recycled through water recovery and purification, so that the consumption of water resources is reduced.
Preferably, the PET/AL/PE composite film is used as a packaging material for prolonging the stability of the product, and the PET/AL/PE composite film packaging materials are all degradable biological materials for improving the environmental protection of the product.
The invention provides a fermentation method of vitamin k 2. The beneficial effects are as follows:
1. according to the invention, the bacillus subtilis is combined with the culture medium containing glucose, peptone, trace elements and vitamins, so that the growth of bacillus natto and the production of vitamin K2 are promoted, the production capacity of the vitamin K2 is enhanced, and the yield and the production efficiency of the vitamin K2 can be further improved.
2. According to the invention, by using a reaction engineering principle, a mathematical modeling and optimizing algorithm accurately controls each parameter, the yield of vitamin K2 is improved to the maximum extent, and then coenzyme Q10 and vitamin D are added to promote the growth of the strain and the synthesis of vitamin K2, so that the yield and the quality can be improved.
3. According to the invention, dimethylbenzene is used as a solvent, soaking and extracting time is controlled, separation and concentration are carried out by utilizing a centrifugal technology and an ultrafiltration membrane, so that energy consumption is reduced, extraction efficiency is improved, and natural flavonoid compounds are used as antioxidants, so that stability of an extracting solution is enhanced, and product quality is maintained.
4. The invention adopts a recycling mode to recycle and reuse waste, reduces environmental burden, selects biodegradable materials to prepare culture medium and equipment, adopts a water circulation system, reduces consumption of plastic and water resources, and improves environmental protection.
Detailed Description
The technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The embodiment of the invention provides a fermentation method of vitamin k2, which comprises the following steps of;
step one, selecting a proper strain, wherein the strain is bacillus subtilis which is a bacterium with stronger heat resistance, has good fermentation capacity, is adaptive to different environmental conditions, and provides a certain stability for the production of vitamin K2;
step two, preparing a fermentation medium, wherein a carbon source selects 20 to 25g/L of glucose, a nitrogen source selects 10 to 12g/L of peptone, and a proper amount of microelements and vitamins are added for promoting the growth of bacillus natto and the production of vitamin K2, the design of the fermentation medium is critical to the growth and metabolic activity of microorganisms, the carbon source and the nitrogen source meet the comprehensive nutrition requirements of the microorganisms in the fermentation process, the utilization efficiency of the strains on the nutrition components in the medium is improved, and the design of the comprehensive medium is beneficial to improving the production efficiency of enzyme-producing strains and maintaining the healthy state of cells;
and thirdly, fermenting, wherein the temperature is maintained between 35 ℃ and 40 ℃, the pH value is maintained between 6.8 and 7.2, the humidity is controlled between 65% and 75%, the oxygen supply is controlled between 0.2vvm and 0.4vvm through the gas flow rate, the optimal growth and enzyme production environment is ensured through a real-time monitoring technology, key parameters in the fermentation process can be accurately regulated and controlled through the real-time monitoring technology, including but not limited to the thallus growth rate, the enzyme production rate, the waste accumulation and the like, and the real-time monitoring technology is to connect various devices and sensors through the technology of the Internet of things, so that the devices and the sensors can communicate and share data with each other. The real-time data flow can be used for monitoring the state of equipment, collecting environmental data and realizing more intelligent decision and control, and each parameter in the fermentation process is accurately controlled by using a reaction engineering principle and a mathematical modeling and optimizing algorithm when fermentation is carried out, so that the biological reaction condition is optimized to the greatest extent and the enzyme production efficiency is improved;
step four, adding proper auxiliary nutrition components such as coenzyme Q10 and vitamin D in the fermentation process for promoting the growth of strains and the synthesis of vitamin K2, wherein the use of the components such as coenzyme Q10 and vitamin D not only helps to increase the concentration of vitamin K2 in fermentation broth, but also can positively influence the metabolic pathway of microorganisms, so that the overall fermentation effect is improved;
and fifthly, extracting by taking dimethylbenzene as a solvent, wherein the soaking time is controlled to be 18-30 hours, the extracting time is controlled to be 3-5 hours, and the dimethylbenzene has high extracting efficiency and selectivity and is relatively environment-friendly. In addition, in the selection of the extraction time, too long or too short time may affect the extraction effect of vitamin K2;
and step six, a column chromatography technology is used, a C18 reverse phase column with the diameter of 10mm and the length of 300mm is used for high-efficiency separation, the solvent gradient is gradually increased from 20% acetonitrile to 80% acetonitrile, the flow rate is 1.0mL/min, impurities are removed, the purification effect is improved, the energy consumption is reduced, the extraction efficiency is improved, and the combination technology can not only effectively reduce the energy consumption, but also improve the purity of the product. The aperture of the ultrafiltration membrane is adjusted according to the size of the target molecule, so as to ensure the efficient concentration of vitamin K2;
step seven, 0.1 to 0.3 percent of natural flavonoid compounds, 0.1 to 0.2 percent of vitamin E as an antioxidant and 0.05 to 0.1 percent of benzoic acid as a preservative are used for enhancing the stability of the extracting solution, the natural flavonoid compounds are used for enhancing the stability of the extracting solution and possibly have the protection effect on vitamin K2, and the compounds have oxidation resistance and can be used for effectively reducing the risk of oxidation of the vitamin K2 in the production process, and the oxidation can lead to the degradation of the vitamin K2, so that the quality and the efficacy of the vitamin K2 are reduced;
and step eight, performing comprehensive quality detection on the vitamin K2 by using a high-resolution analysis technology, including but not limited to a liquid chromatography-mass spectrometry technology, nuclear magnetic resonance and the like, so as to ensure that the purity, structure and content of the product meet the expected requirements. The detailed quality detection is helpful for guaranteeing the compliance and stability of vitamin K2, and simultaneously provides data support for further optimization of products;
in the first step, a proper strain is selected, wherein the strain is bacillus subtilis;
specifically, bacillus subtilis with strong heat resistance, excellent fermentation capability and high adaptability to environmental conditions is selected as a microbial basis of the experiment, so that the repeatability and controllability of the experiment are improved, and favorable conditions are provided for efficient production of vitamin K2.
In the second step, the trace elements are 0.3 to 0.5 g of calcium, 0.01 to 0.02 g of zinc and 0.1 to 0.2 g of magnesium, and the vitamin is vitamin D, so that the utilization efficiency of the strain on nutrient components in a culture medium is improved, and the production capacity of vitamin K2 is enhanced;
specifically, the production capacity of vitamin K2 is further enhanced by optimizing the cell metabolism way and improving the utilization efficiency of nutrient components in the culture medium, the important roles of microelements and vitamins in cell biology are fully utilized, and a solid scientific basis is provided for the application of the strain in industrial production.
In the third step, the reaction engineering principle is used during fermentation, and each parameter in the fermentation process is precisely controlled through mathematical modeling and optimization algorithm, so that the yield of vitamin K2 is improved to the maximum extent;
specifically, by establishing a mathematical model for the growth and metabolism of microorganisms, key influencing factors of the biological processes can be more comprehensively known, so that the reaction conditions can be pertinently adjusted, the mathematical modeling and optimizing algorithm can accurately control the key parameters such as temperature, pH value, oxygen concentration and the like in the fermentation process, the growth and the product generation of microorganisms in the most appropriate environment are ensured, a scientific and intelligent method is provided for the production of vitamin K2, the fermentation process is more controllable and efficient, and the yield is improved to the maximum extent.
In the third step, renewable energy sources such as solar energy and wind energy are used in the fermentation process, so that the dependence on the traditional energy sources is reduced, and the environmental impact is reduced;
specifically, replacing traditional energy sources, such as fossil fuels, reduces greenhouse gas emissions and over-exploitation of natural resources. This helps to reduce the overall environmental impact of the fermentation process, making the production process more environmentally friendly and sustainable, and the renewable nature of solar and wind energy makes it relatively stable, not being limited to geographical and climatic conditions as some traditional energy sources. This ensures a reliable supply of energy, which contributes to an improved stability of the fermentation process and to a continuity of production.
In the eighth step, the high-resolution analysis technology includes high performance liquid chromatography and mass spectrometry analysis technology, which can perform comprehensive mass detection on the vitamin K2;
specifically, the high performance liquid chromatography and the mass spectrometry are widely applied to industries such as medicines, foods and the like, and can realize comprehensive quality detection of the vitamin K2 by combining analysis of the high performance liquid chromatography and the mass spectrometry. This includes accurate determination of the various components in the product, including possible impurities, reaction products and other organic or inorganic compounds. Through high-resolution analysis, the purity and consistency of the product can be ensured, the quality standard can be met, the analysis technologies are not only used for quality detection of vitamin K2, but also can be used for real-time monitoring in the production process, and through timely detection of possible problems, a producer can conduct real-time process control and improvement, and the stability and controllability of the quality of the product are ensured.
In the second step, a biodegradable material is used for preparing a fermentation medium and production equipment, so that the dependence of the whole production process on plastics and other non-degradable materials is reduced;
in particular, the use of biodegradable materials allows for a higher reproducibility of the fermentation medium. This means that the raw materials used in the production process come from renewable resources, which helps to reduce the dependence on limited resources and reduce the impact of production on the environment.
In the third step, a water circulation system is used, so that the water consumption is reduced to the greatest extent, and the consumption of water resources is reduced through the recovery, purification and reutilization of water;
specifically, the waste generated in the fermentation process is recovered and reused, the burden on the environment is reduced, the biodegradable material is used for preparing the fermentation medium and the production equipment, the dependence of the whole production process on plastics and other non-degradable materials is reduced, and adverse effects on the environment in the production process can be furthest reduced through effective environmental protection measures such as composting of organic waste, treatment and recovery of waste water and the like.
The PET/AL/PE composite film is used as a packaging material for prolonging the stability of the product, and the PET/AL/PE composite film packaging materials are degradable biological materials for improving the environmental protection property of the product and prolonging the stability of the product;
specifically, the degradable packaging material, the recyclable material and the optimization of the packaging process not only accord with the concept of sustainable development, but also help to prolong the stability of the product, and help to lighten the burden of the whole supply chain on the environment by reducing the influence of the packaging material on the product.
The following description is made in connection with the specific implementation steps:
embodiment one;
a fermentation method of vitamin k2, comprising the steps of;
step one, selecting a proper strain, wherein the strain is bacillus subtilis;
step two, selecting 25g/L of glucose as a carbon source, and selecting 12g/L of peptone as a nitrogen source, wherein the microelements comprise 0.5 g of calcium, 0.02 g of zinc and 0.1 g of magnesium;
maintaining the temperature at 35 ℃, the pH value at 6.8, the humidity at 65%, and the oxygen supply at 0.2vvm through the gas flow rate;
step four, adding proper auxiliary nutrients such as coenzyme Q10 and vitamin D in the fermentation process, wherein the auxiliary nutrients are used for promoting the growth of the strain and the synthesis of vitamin K2;
step five, extracting by using dimethylbenzene as a solvent, wherein the soaking time is controlled to be 30 hours, and the extracting time is controlled to be 5 hours;
step six, using column chromatography technology for high-efficiency separation of C18 reverse phase column with diameter of 10mm and length of 300mm, gradually increasing solvent gradient from 20% acetonitrile to 80% acetonitrile, and flow rate of 1.0mL/min for removing impurities and improving purification effect;
in the seventh step, 0.3% of natural flavonoid, 0.1% of vitamin E as an antioxidant and 0.05% of benzoic acid as a preservative are used for enhancing the stability of the extracting solution;
and step eight, carrying out comprehensive quality detection on the vitamin K2 by using a high-resolution analysis technology.
Embodiment two;
the difference between this embodiment and the first embodiment is that:
in the second step, 30g/L of glucose is selected as a carbon source, 15g/L of peptone is selected as a nitrogen source, and 0.4 g of calcium, 0.03 g of zinc and 0.1 g of magnesium are used as microelements.
In step three, the temperature was maintained at 40 degrees celsius, the pH was maintained at 7.3, the humidity was controlled at 75%, and the oxygen supply was controlled at 0.2vvm by gas flow rate.
In the fifth step, xylene is selected as a solvent for extraction, the soaking time is controlled to be 24 hours, and the extraction time is controlled to be 4 hours.
The specific experiment:
the purpose of the experiment is as follows: the influence of different fermentation conditions on the yield and quality of vitamin K2 is explored, and a reference is provided for optimizing a fermentation process.
Experimental materials: bacillus subtilis strain, fermentation medium, xylene, C18 reverse phase column, acetonitrile, natural flavonoid, vitamin E, benzoic acid, high performance liquid chromatograph, mass spectrometer, etc.
The experimental method comprises the following steps:
1. the Bacillus subtilis strain was precultured in a fermentation medium containing glucose, peptone, calcium, zinc, magnesium, vitamin D, etc., with an inoculum size of 2%, an initial pH of 7.2, and a liquid loading of 16mL/50mL.
2. Inoculating the preculture solution into four different fermentation tanks A, B, C, D respectively, wherein the capacity of each fermentation tank is 5L, the fermentation time is 96h, coenzyme Q10 and vitamin D are added as auxiliary nutritional components in the fermentation process, and the fermentation conditions of the different fermentation tanks are shown in the following table:
| fermentation tank | Temperature (. Degree. C.) | pH | Humidity (%) | Oxygen flow rate (vvm) |
| A | 35 | 6.8 | 65 | 0.2 |
| B | 35 | 7.2 | 75 | 0.4 |
| C | 40 | 6.8 | 75 | 0.2 |
| D | 40 | 7.2 | 65 | 0.4 |
After fermentation, extracting vitamin K2 in the fermentation liquid by using dimethylbenzene respectively, wherein the extraction conditions of different fermentation tanks are shown in the following table;
| fermentation tank | Soaking time (h) | Extraction time (h) |
| A | 18 | 3 |
| B | 24 | 4 |
| C | 30 | 5 |
| D | 24 | 4 |
Performing column chromatography with C18 reverse phase column, gradually increasing solvent gradient from 20% acetonitrile to 80% acetonitrile at flow rate of 1.0mL/min, removing impurities and improving purification effect, and concentrating with ultrafiltration membrane to obtain vitamin K2 extractive solution.
Natural flavonoid compounds, vitamin E and benzoic acid are added into the extracting solution to serve as antioxidants and preservatives, stability of the extracting solution is enhanced, and the adding amounts of different fermentation tanks are shown in the following table:
| fermentation tank | Natural flavonoid (%) | Vitamin E (%) | Benzoic acid (%) |
| A | 0.1 | 0.1 | 0.05 |
| B | 0.2 | 0.2 | 0.1 |
| C | 0.3 | 0.1 | 0.05 |
| D | 0.2 | 0.1 | 0.1 |
And (3) measuring the content of vitamin K2 in the extracting solution by using a high performance liquid chromatography and a mass spectrometry, and identifying the structure of the vitamin K2 in the extracting solution by using a nuclear magnetic resonance spectrometer to obtain a quality detection result of a final product.
Experimental results: according to the experimental method, comparative data of the yield and quality of vitamin K2 under different fermentation conditions are obtained as shown in the following table:
experimental analysis: from the experimental results, the following conclusions can be drawn: different fermentation conditions have certain influence on the yield and quality of the vitamin K2, but the influence is not great, so that the bacillus subtilis strain has strong adaptability to the fermentation conditions, a certain stability is provided for the production of the vitamin K2, the structures of the vitamin K2 obtained by all fermentation tanks are consistent with those of standard substances, the structure of the vitamin K2 is not changed in the extraction and purification processes, the activity and the efficacy of the product are ensured, the yield and the production efficiency of the vitamin K2 can be further improved, and the quality of the product is maintained.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations may be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (9)
1. A fermentation method of vitamin k2, comprising the steps of;
step one, selecting a proper strain, wherein the strain is bacillus subtilis;
step two, preparing a fermentation culture medium, wherein a carbon source selects 20 to 25g/L of glucose, a nitrogen source selects 10 to 12g/L of peptone, and a proper amount of trace elements and vitamins are added for promoting the growth of bacillus subtilis and the production of vitamin K2;
step three, fermentation is carried out, wherein the temperature is maintained between 35 ℃ and 40 ℃, the pH value is maintained between 6.8 and 7.2, the humidity is controlled between 65% and 75%, the oxygen supply is controlled between 0.2 and 0.4vvm through the gas flow rate, and the optimal growth and enzyme producing environment is ensured through a real-time monitoring technology;
step four, adding proper auxiliary nutrients such as coenzyme Q10 and vitamin D in the fermentation process, wherein the auxiliary nutrients are used for promoting the growth of the strain and the synthesis of vitamin K2;
step five, extracting by using dimethylbenzene as a solvent, wherein the soaking time is controlled to be 18 to 30 hours, and the extracting time is controlled to be 3 to 5 hours;
step six, using column chromatography technology, carrying out high-efficiency separation by using a C18 reverse phase column with the diameter of 10mm and the length of 300mm, gradually increasing the solvent gradient from 20% acetonitrile to 80% acetonitrile, and enabling the flow rate to be 1.0mL/min so as to remove impurities and improve the purification effect;
step seven, using 0.1 to 0.3 percent of natural flavonoid compounds, 0.1 to 0.2 percent of vitamin E as an antioxidant and 0.05 to 0.1 percent of benzoic acid as a preservative for enhancing the stability of the extracting solution;
and step eight, carrying out comprehensive quality detection on the vitamin K2 by using a high-resolution analysis technology.
2. The method for fermenting vitamin K2 according to claim 1, wherein in the first step, the bacillus subtilis is a bacterium with strong heat resistance, has good fermentation capacity, is adaptive to different environmental conditions, and provides a certain stability for the production of vitamin K2.
3. The method for fermenting vitamin K2 according to claim 1, wherein in the second step, the trace elements are 0.3 to 0.5 g of calcium, 0.01 to 0.02 g of zinc and 0.1 to 0.2 g of magnesium, and the vitamin is vitamin D, so as to improve the utilization rate of the strain on nutrient components in the culture medium and enhance the production capacity of vitamin K2.
4. The method for fermenting vitamin K2 according to claim 1, wherein in the third step, the reaction engineering principle is used in the fermentation process, and each parameter in the fermentation process is precisely controlled by mathematical modeling and optimization algorithm, so that the yield of vitamin K2 is improved to the maximum extent.
5. The method for fermenting vitamin k2 according to claim 1, wherein in the third step, renewable energy sources such as solar energy and wind energy are used in the fermentation process, so that the dependence on traditional energy sources is reduced, and the environmental impact is reduced.
6. The method of claim 1, wherein in step eight, the high resolution analysis technique comprises high performance liquid chromatography and mass spectrometry analysis techniques that provide comprehensive mass detection of vitamin K2.
7. The method of claim 1, wherein in the second step, the biodegradable material is used to prepare the fermentation medium and the production equipment, so that the dependence of the whole production process on plastics and other non-degradable materials is reduced.
8. The method for fermenting vitamin k2 according to claim 1, wherein in the third step, a water circulation system is used to minimize water consumption, and water resource consumption is reduced by recovering and purifying water for reuse.
9. The method for fermenting vitamin k2 according to claim 1, wherein the PET/AL/PE composite film is used as a packaging material for prolonging the stability of the product, and the PET/AL/PE composite film packaging materials are degradable biological materials for improving the environmental protection property of the product.
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