CN111996219B - Fermentation method of coenzyme Q10 - Google Patents

Fermentation method of coenzyme Q10 Download PDF

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CN111996219B
CN111996219B CN202011030403.2A CN202011030403A CN111996219B CN 111996219 B CN111996219 B CN 111996219B CN 202011030403 A CN202011030403 A CN 202011030403A CN 111996219 B CN111996219 B CN 111996219B
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段志钢
王平
尹贵超
王成
王淑琳
张军剪
王云
牛霄亮
李景
李默晗
高雨宁
丁宝娟
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Abstract

The invention discloses a fermentation method of coenzyme Q10, belonging to the field of microbial fermentation. The fermentation control process obviously enhances the growth and metabolic capacity of the coenzyme Q10 producing strain, obviously increases the strain amount, increases the fermentation unit of the coenzyme Q10 quickly, ensures that the tank-placing potency of the coenzyme Q10 reaches more than 4000mg/L, obviously prolongs the fermentation period, easily filters and collects thalli, greatly increases the batch yield of the coenzyme Q10, and obviously reduces the fermentation cost of the coenzyme Q10.

Description

Fermentation method of coenzyme Q10
Technical Field
The invention relates to a fermentation method of coenzyme Q10, belonging to the field of microbial fermentation.
Background
Coenzyme Q10(Ubiquinone-10) is a naturally occurring fat-soluble quinone compound, also known as decene quinone, Ubiquinone-10, and has the chemical name: 2, 3-dimethoxy-5-methyl-6-deca-isoprenyl-1, 4-benzoquinone (2,3-dimethoxy-5-methyl-6-decaprenyl-1,4-benzoquinone), the molecular formula is: c59H90O4The molecular weight is: 863.37, the structural formula is:
Figure BDA0002703457130000011
the compound is yellow to orange crystalline powder, is odorless and tasteless, is easily soluble in benzene, acetone and ether, is hardly soluble in water and methanol, is unstable under direct sunlight, is widely distributed in nature, mainly exists in yeast, plant leaves, seeds and animal heart, liver and kidney cells, particularly in mammal biomembranes rich in mitochondria, has higher content in human heart, liver, kidney and pancreas, and can activate cell respiration, accelerate the generation of ATP with high energy, strengthen the myocardial metabolism function, improve the heart beating efficiency, treat cardiovascular diseases, such as ischemic heart disease, rheumatic heart disease, myocarditis, angina pectoris, arrhythmia, hypertension and the like; the coenzyme Q10 can also inhibit the peroxidation of mitochondria, protect the structural integrity of a biological membrane, and has the functions of improving the immunity of a human body, delaying senility, enhancing the vitality of the human body and the like. The coenzyme Q10 has obvious curative effect on the aspects of comprehensively treating cardiovascular diseases and cancers, preventing dental diseases and the like. Meanwhile, effective applications in the fields of health products and cosmetics have shown its strong demand.
The present technique for industrially producing coenzyme Q10 at home and abroad is a microbial fermentation method. Rhodobacter sphaeroides (rhodobacter sphaeroides) is used as coenzyme Q10 zymogen, and the zymogen is inoculated by slant or shake flask bacteria liquid for three-stage fermentation. The culture medium is glucose, corn steep liquor powder, ammonium sulfate, magnesium sulfate, ferric chloride, sodium chloride, monosodium glutamate, potassium dihydrogen phosphate, trace elements, etc. commonly used, glucose, ammonia water and phosphorus elements are supplemented in the fermentation process, the temperature of the fermentation tank is 32-35 ℃, and the pH value of the fermentation tank is controlled to be 6.5-7.0 by using the ammonia water. For example, the industrial production method of coenzyme Q10 in the Chinese patent with the publication number CN102168115 is described as follows: the fermentation unit of coenzyme Q10 is improved by adding key storage factors, controlling the content of dissolved phosphorus, controlling the content of amino nitrogen, supplementing 3 times of concentrated fermentation base materials and the like, and the fermentation unit reaches 3400 mg/L; the fermentation process and the control strategy of coenzyme Q10 in the Chinese patent with the publication number of CN104561154 are described as follows: the strain shape transition period in the fermentation process is taken as a time point for adjusting process parameters, and the fermentation unit of coenzyme Q10 is improved by adjusting methods such as ventilation volume, tank pressure, stirring speed and the like of a fermentation tank, wherein the fermentation unit reaches 3500 mg/L; the Chinese patent with publication number CN105483171 describes a production method for improving the industrial yield of coenzyme Q10: the fermentation unit of coenzyme Q10 is increased by controlling the concentration of ferrous ions in a seed culture medium, the fermentation unit reaches 3382mg/L, and the fermentation technology of coenzyme Q10 is relatively mature, but the existing industrial production method for producing coenzyme Q10 by microbial fermentation has the problem of low fermentation unit.
Disclosure of Invention
In view of the above, the present invention provides a method for producing coenzyme Q10 by fermentation, comprising: (1) continuously supplementing ammonium ions in the fermentation process of coenzyme Q10, and controlling different ammonium ion concentrations in different fermentation periods; (2) adding lysine salt into a coenzyme Q10 fermentation medium, and supplementing sodium sulfate or potassium sulfate in the fermentation process of adding lysine salt 3 times and supplementing coenzyme Q10 3 times in the fermentation process. After the fermentation control process is used, the fermentation unit of the coenzyme Q10 is obviously improved, the dry weight of thalli is obviously increased, the fermentation period is obviously prolonged, the yield of the coenzyme Q10 is greatly increased, and the fermentation cost of the coenzyme Q10 is obviously reduced.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for fermenting coenzyme Q10,
1) mother bottle culture: diluting coenzyme Q10 production strains to 1200 strains/mL, inoculating 0.5mL to a slant culture medium, placing the slant culture medium in an incubator at the temperature of 30-34 ℃ and the relative humidity of 40-60% for culturing for 4-8 days, selecting 2-5 colonies, inoculating the colonies in a mother bottle culture medium, placing the mother bottle culture medium in a shaker at the temperature of 30-34 ℃, the relative humidity of 40-60% and the rotation speed of 200-250rpm for culturing for 24-32 hours to obtain mother bottle strains;
the formula of the slant culture medium is as follows: 1.0-4.0g/L glucose, 0.5-2.0g/L yeast powder, 0.2-1.0g/L peptone, 0.5-2.0g/L sodium chloride, 15-25g/L agar powder, pH6.8-7.2, sterilizing at 121 deg.C for 20-25 min;
the mother flask culture medium formula is as follows: 3.0-6.0g/L of glucose, 0.2-0.5g/L of yeast powder, 0.1-0.5g/L of peptone, 0.5-1.0g/L of corn steep liquor, 0.5-2.0g/L of ammonium sulfate, 0.4-2g/L of monosodium glutamate, 0.5-2.0g/L of sodium chloride, 1.0-3.0g/L of magnesium sulfate, 0.1-0.5g/L of potassium dihydrogen phosphate, 0.1-0.5g/L of dipotassium hydrogen phosphate, 0.1-0.5g/L of ferric chloride, 2.0-8.0g/L of calcium carbonate, 1.0-3.0mg/L of thiamine hydrochloride, 2-5mg/L of nicotinic acid, 0.05-0.2mg/L of biotin, 6.5-7.0 of pH value, and 20-25min of sterilization at 121 ℃.
2) First-level seeding tank culture: inoculating the strain in the mother bottle to a culture medium in a first-stage seed tank according to the inoculation amount of 1-5 per mill, culturing for 28-36h at 30-35 ℃, the tank pressure of 0.03-0.05MPa, the aeration ratio of 0.3-0.6 and the stirring rotation speed of 140 plus materials of 180rpm to obtain a first-stage seed solution with the glucose residual amount of 0.5-1.0g/L, uniform thallus shape and good sterility;
the formula of the culture medium in the first-level seeding tank is as follows: glucose 4.0-8.0g/L, yeast powder 0.3-0.6g/L, peptone 0.1-0.4g/L, corn steep liquor 0.6-1.2g/L, ammonium sulfate 0.8-2.5g/L, monosodium glutamate 0.4-2g/L, sodium chloride 0.5-2.0g/L, magnesium sulfate 1.5-4.0g/L, potassium dihydrogen phosphate 0.1-0.5g/L, dipotassium hydrogen phosphate 0.1-0.5g/L, ferric chloride 0.1-0.5g/L, calcium carbonate 2.0-8.0g/L, thiamine hydrochloride 1.0-3.0mg/L, nicotinic acid 2.0-5.0mg/L, biotin 0.05-0.2mg/L, pH6.5-7.0, and sterilizing at 121 ℃ for 25-30 min.
3) Secondary seeding tank culture: inoculating the first-stage seed liquid to a culture medium in a second-stage seed tank according to the inoculation amount of 8-15%, culturing for 16-24h at 30-35 ℃, the tank pressure of 0.03-0.05MPa, the aeration ratio of 0.4-0.7 and the stirring speed of 120-160rpm to obtain the second-stage seed liquid with the glucose residual amount of 0.4-0.8g/L, uniform thallus shape and good sterility;
the formula of the culture medium in the secondary seeding tank is as follows: 6.0-12g/L glucose, 0.3-0.6g/L yeast powder, 0.8-1.5g/L corn steep liquor, 1.0-3.5g/L ammonium sulfate, 0.8-3g/L monosodium glutamate, 0.5-2.0g/L sodium chloride, 1.5-4.0g/L magnesium sulfate, 0.1-0.5g/L potassium dihydrogen phosphate, 0.1-0.5g/L dipotassium hydrogen phosphate, 0.1-0.5g/L ferric chloride, 2.0-8.0g/L calcium carbonate, 1.0-3.0mg/L thiamine hydrochloride, 4.0-8.0mg/L nicotinic acid, 0.08-0.3mg/L biotin, 6.5-7.0 pH, and 25-30min sterilization at 121 ℃.
4) Culturing in a fermentation tank: inoculating the second-stage seed liquid into a culture medium in a fermentation tank according to the inoculation amount of 15-25%, culturing at 32-35 ℃, the tank pressure of 0.03-0.06MPa, the aeration ratio of 0.6-0.8 and the stirring speed of 110-140rpm until the glucose consumption rate of thalli is obviously reduced, and stopping fermentation when thalli are partially autolyzed;
the culture medium formula in the fermentation tank is as follows: 10-15g/L of glucose, 1.5-2.5g/L of corn steep liquor, 1.5-3.5g/L of ammonium sulfate, 1.0-3g/L of monosodium glutamate, 0.5-2.0g/L of sodium chloride, 3.0-6.0g/L of magnesium sulfate, 0.1-0.5g/L of potassium dihydrogen phosphate, 0.1-0.5g/L of dipotassium hydrogen phosphate, 0.2-1.0g/L of ferric chloride, 1.0-3.0g/L of calcium carbonate, 2.0-5.0mg/L of thiamine hydrochloride, 6.0-12mg/L of nicotinic acid and 0.1-0.5mg/L of biotin; lysine salt 0.001-5g/L, preferably 0.01-0.1 g/L; sterilizing at 121 deg.C for 25-30min at pH of 6.3-6.8.
Fermentation tank process control measures:
a) and (3) supplementing the ammonium salt aqueous solution in the following manner: when the fermentation period is 1-20h, the ammonium ion concentration is supplemented to the fermentation tank to be 0.8-1.2 g/L; when the fermentation period is 21-40h, the ammonium ion concentration is 1.2-1.6 g/L; when the fermentation period is 41-65h, the ammonium ion concentration is supplemented to the fermentation tank to be 1.6-2.0 g/L; when the fermentation period is 66-80h, the ammonium ion concentration is 1.2-1.6 g/L; the fermentation period is 81 h-when the fermentation tank is placed, the ammonium ion concentration is supplemented to the fermentation tank to be 0.6-1.2 g/L;
b) and (3) supplementing the lysine salt aqueous solution in the following way: when the fermentation period is 20-28h, the lysine salt concentration in the fermentation tank is 0.01-0.1 g/L; when the fermentation period is 40-48h, the lysine salt concentration in the fermentation tank is 0.04-0.12 g/L; when the fermentation period is 60-68h, the lysine salt concentration in the fermentation tank is 0.005-0.05 g/L;
c) and (3) adding the sulfate aqueous solution, wherein the adding mode of the sulfate aqueous solution is as follows: when the fermentation period is 36-70h, preferably 48-58h, the lysine salt sulfate concentration is 0.2-0.8g/L when the lysine salt sulfate is supplemented into the fermentation tank;
d) continuously supplementing 50-60% glucose solution in the fermentation tank during the culture process to control the residual glucose amount in the fermentation tank at 8-15 g/L; continuously supplementing 13-16% potassium dihydrogen phosphate solution during the culture process to control the phosphorus residue in the fermentation tank at 0.15-0.25 g/L; ammonia water with concentration of 18-22% is continuously added during the culture process, so that the pH value in the fermentation tank is controlled at 6.5-7.0.
Further, the aqueous ammonium salt solution is an aqueous ammonium sulfate, ammonium chloride, ammonium carbonate or ammonium bicarbonate solution, and the concentration of the aqueous ammonium salt solution is 5% -80%, preferably 20% -60%, more preferably 30% -50%, and most preferably 35% -45%;
the lysine salt aqueous solution is lysine hydrochloride, lysine sulfate or lysine phosphate aqueous solution, and the concentration of the lysine salt aqueous solution is 1-30%, preferably 2-20%, more preferably 3-10%;
the sulfate aqueous solution is sodium sulfate or potassium sulfate aqueous solution, and the concentration of the sulfate aqueous solution is 10% -60%, preferably 20% -50%, more preferably 30% -40%.
The invention has the beneficial effects that: in the test process, the requirement of the fermentation process of the coenzyme Q10 producing strain on ammonium ions is surprisingly found to be very obvious, and although ammonia water is supplemented in the fermentation process to control the pH of the fermentation liquor to be 6.5-7.0, the ammonium ions provided by the ammonia water are far from insufficient, so the method adds the operation of continuously supplementing ammonium sulfate in the fermentation process of the coenzyme Q10, and controls different ammonium ion concentrations in different fermentation periods to enable the growth and metabolism of the coenzyme Q10 producing strain to reach the optimal state.
Adding lysine salt into coenzyme Q10 fermentation medium, and supplementing lysine salt 3 times in the fermentation process; sodium sulfate is added once in the middle of the fermentation of the coenzyme Q10, so that the growth of the coenzyme Q10 producing bacteria is obviously promoted, and the bacteria are stout.
By the technical means, the growth and metabolic capacity of the coenzyme Q10 producing strain are obviously enhanced; the bacterial quantity is obviously increased by about 10 percent; the fermentation unit of the coenzyme Q10 is increased quickly, the tank-discharging titer of the coenzyme Q10 reaches more than 4000mg/L, and is increased by about 20 percent compared with the coenzyme Q10 before the process is used; the fermentation period is obviously prolonged, the batch yield of the coenzyme Q10 is greatly increased, the thalli are easy to filter and collect, and the fermentation cost of the coenzyme Q10 is obviously reduced.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the embodiment of the present invention, the method can be applied to any species that produces coenzyme Q10, and as a specific embodiment, the species used is rhodobacter sphaeroides (Rhodobacterphoroides).
Example 1
Seed culture: diluting coenzyme Q10 production strains in a glycerinum tube to a proper concentration, inoculating the strains to a slant culture medium of an eggplant bottle, and culturing the slant in an incubator at the temperature of 32 +/-0.5 ℃ and the relative humidity of 40-60% for 6 days. After the slant culture of the eggplant bottle is finished, 3 colonies are picked by an inoculating needle and inoculated in a 500mL triangular flask filled with 60mL culture medium, and a mother bottle is placed in a shaking table with the temperature of 32 +/-0.5 ℃, the relative humidity of 40-60% and the rotating speed of 220rpm for culture for 30 hours and then used as a mother bottle seed to inoculate a primary seed tank. The eggplant bottle slant culture medium comprises the following components: 2g/L glucose, 1.5g/L yeast powder, 0.5g/L peptone, 1.5g/L sodium chloride, 15g/L agar powder, pH6.85, and sterilizing at 121 + -0.5 deg.C for 20 min. The mother flask culture medium formula is as follows: 5g/L glucose, 0.4g/L yeast powder, 0.3g/L peptone, 0.7g/L corn steep liquor, 1.2g/L ammonium sulfate, 1.5g/L monosodium glutamate, 1.5g/L sodium chloride, 2.5g/L magnesium sulfate, 0.4g/L potassium dihydrogen phosphate, 0.4g/L dipotassium hydrogen phosphate, 0.2g/L ferric chloride, 5.0g/L calcium carbonate, 2.0mg/L thiamine hydrochloride, 4.0mg/L nicotinic acid, 0.1mg/L biotin, pH6.8, and sterilization at 121 +/-0.5 ℃ for 20 min.
First-stage seeding tank expanded culture: inoculating the cultured mother bottle seeds into a first-stage seed tank according to the inoculation amount of 2 thousandth, wherein the volume of a culture medium of the first-stage seed tank is 800L, and the culture conditions of the first-stage seed tank are as follows: the temperature of the tank is 32 plus or minus 0.5 ℃, the pressure of the tank is 0.04 plus or minus 0.005MPa, the aeration ratio is 0.5, the stirring speed is 160rpm, after culturing for 30 hours, the residual glucose content in the seed liquid is reduced to 0.5g/L, the thallus shape is uniform, the sterility is good, namely, the second-stage seed tank is inoculated according to 10 percent of inoculation amount, the volume of the feed liquid in the second-stage seed tank after inoculation is 8m3The culture conditions of the secondary seeding tank are as follows: the temperature of the tank is 32 plus or minus 0.5 ℃, the pressure of the tank is 0.04 plus or minus 0.005MPa, the aeration ratio is 0.55, the stirring speed is 140rpm, the residual glucose content in the secondary seed liquid is reduced to 0.6g/L after 20 hours of culture, the shape of the thalli is uniform, the bacteria is good, and the fermentation tank can be transplanted. The formula of the first-level seeding tank culture medium is as follows: 5g/L glucose, 0.5g/L yeast powder, 0.2g/L peptone, 0.8g/L corn steep liquor, 1.3g/L ammonium sulfate, 1.5g/L monosodium glutamate, 1.8g/L sodium chloride, 3.0g/L magnesium sulfate, 0.4g/L potassium dihydrogen phosphate, 0.4g/L dipotassium hydrogen phosphate, 0.2g/L ferric chloride, 6g/L calcium carbonate, 1.5mg/L thiamine hydrochloride, 3.5mg/L nicotinic acid, 0.05mg/L biotin, 6.6 pH6, and sterilization is carried out at 121 +/-0.5 ℃ for 30 min. The formula of the culture medium of the secondary seeding tank is as follows: 10g/L of glucose, 0.4g/L of yeast powder, 1.2g/L of corn steep liquor, 2.0g/L of ammonium sulfate, 2.5g/L of monosodium glutamate, 1.8g/L of sodium chloride, 3.5g/L of magnesium sulfate, 0.4g/L of monopotassium phosphate, 0.4g/L of dipotassium phosphate, 0.3g/L of ferric chloride, 4g/L of calcium carbonate, 2.5mg/L of thiamine hydrochloride, 5.0mg/L of nicotinic acid, 0.15mg/L of biotin, pH6.7, and sterilization is carried out at the temperature of 121 +/-0.5 ℃ for 30 min.
Culturing in a fermentation tank: inoculating the cultured seed liquid of the second-stage seed tank into a fermentation tank according to the inoculation amount of 20%, wherein the volume of the liquid in the fermentation tank after inoculation is 40m3The culture conditions of the fermentation tank are as follows: the tank temperature is 33 plus or minus 0.5 ℃, the tank pressure is 0.05 plus or minus 0.005MPa, the aeration ratio is 0.65, and the stirring speed is 120 rpm. The formula of the fermentation tank culture medium is as follows: 12g/L glucose, 2.0g/L corn steep liquor, 2.5g/L ammonium sulfate, 2.5g/L monosodium glutamate, 2.0g/L sodium chloride, 5.5g/L magnesium sulfate, 0.3g/L potassium dihydrogen phosphate, 0.3g/L dipotassium hydrogen phosphate, 0.8g/L ferric chloride, 1.5g/L calcium carbonate, 4.0mg/L thiamine hydrochloride, 8.0mg/L nicotinic acid and 0.25mg/L biotin; lysine hydrochloride 0.02g/L, pH6.4, 121 + -0.5 deg.C sterilization for 30 min.
Fermentation tank process control measures: continuously supplementing 55% glucose solution in the fermentation tank during the culture process, controlling the residual glucose amount in the fermentation tank at 10 + -1 g/L, and stopping supplementing glucose 2 hr before tank placement; continuously supplementing 15% potassium dihydrogen phosphate solution during the culture process, and controlling the phosphorus residue in the fermentation tank at 0.2 + -0.05 g/L; ammonia water with the concentration of 20% is continuously supplemented in the culture process, and the pH value of the fermentation tank is controlled to be 6.7 +/-0.1. Ammonium sulfate solution with the concentration of 35% is continuously supplemented in the fermentation process, and the concentration of ammonium ions in the fermentation tank is controlled as follows: the fermentation period is 1-20h, and the concentration of ammonium ions is 1.1 +/-0.05 g/L; the fermentation period is 21-40h, and the concentration of ammonium ions is 1.4 +/-0.05 g/L; the fermentation period is 41-65h, and the concentration of ammonium ions is 1.8 +/-0.05 g/L; the fermentation period is 66-80h, and the concentration of ammonium ions is 1.3 +/-0.05 g/L; the fermentation period is 81 h-the tank is placed, and the concentration of ammonium ions is 0.8 +/-0.05 g/L.
In the fermentation process, a lysine salt solution with the concentration of 5% is supplemented for 3 times, and the supplementing mode is as follows: supplementing lysine hydrochloride with final concentration of 0.02g/L in a fermentation period of 25 h; supplementing lysine hydrochloride with final concentration of 0.06g/L at fermentation period of 44 h; supplementing lysine hydrochloride with final concentration of 0.01g/L in a fermentation period of 62 h; and supplementing a sodium sulfate solution with the concentration of 35% in the fermentation process, wherein the supplementing mode is as follows: sodium sulfate with final concentration of 0.3g/L is supplemented during the fermentation period of 55 h.
After the fermentation tank is cultured for 98 hours, the glucose consumption rate of thalli is obviously reduced, the thalli is partially autolyzed, the fermentation tank is stopped and put into the tank, the titer of put into the tank of Q10 is 4035mg/L, and the dry weight of the thalli is 103 g/L.
Example 2
Seed culture: diluting coenzyme Q10 production strains in a glycerinum pipe to a proper concentration, inoculating the strains to a slant culture medium of an eggplant bottle, and culturing the slant in an incubator at the temperature of 32 +/-0.5 ℃ and the relative humidity of 40-60% for 5 days. After the slant culture of the eggplant bottle is finished, 4 colonies are picked by an inoculating needle and inoculated in a 500mL triangular flask filled with 60mL culture medium, and a mother bottle is placed in a shaking table with the temperature of 32 +/-0.5 ℃, the relative humidity of 40-60% and the rotating speed of 220rpm for culture for 28h and then used as a mother bottle seed to inoculate a primary seed tank. The eggplant bottle slant culture medium comprises the following components: 3g/L of glucose, 1.6g/L of yeast powder, 0.4g/L of peptone, 1.5g/L of sodium chloride, 15g/L of agar powder, pH6.80 and sterilization at the temperature of 121 +/-0.5 ℃ for 20 min. The mother flask culture medium formula is as follows: 4g/L of glucose, 0.5g/L of yeast powder, 0.2g/L of peptone, 0.6g/L of corn steep liquor, 1.3g/L of ammonium sulfate, 1.6g/L of monosodium glutamate, 1.5g/L of sodium chloride, 2.8g/L of magnesium sulfate, 0.4g/L of potassium dihydrogen phosphate, 0.4g/L of dipotassium hydrogen phosphate, 0.2g/L of ferric chloride, 5.0g/L of calcium carbonate, 2.0mg/L of thiamine hydrochloride, 4.0mg/L of nicotinic acid, 0.1mg/L of biotin, pH6.85, and sterilization at the temperature of 121 +/-0.5 ℃ for 20 min.
First-stage seeding tank expanded culture: inoculating the cultured mother bottle seeds into a first-stage seed tank according to the inoculation amount of 1 thousandth, wherein the volume of a culture medium in the first-stage seed tank is 800L, and the culture conditions of the first-stage seed tank are as follows: the temperature of the tank is 32 plus or minus 0.5 ℃, the pressure of the tank is 0.04 plus or minus 0.005MPa, the aeration ratio is 0.5, the stirring speed is 160rpm, after culturing for 32 hours, the residual glucose content in the seed liquid is reduced to 0.4g/L, the thallus shape is uniform, the sterility is good, namely, the second-stage seed tank is inoculated according to 10 percent of inoculation amount, the volume of the feed liquid in the second-stage seed tank after inoculation is 8m3The culture conditions of the secondary seeding tank are as follows: the temperature of the tank is 32 plus or minus 0.5 ℃, the pressure of the tank is 0.04 plus or minus 0.005MPa, the aeration ratio is 0.55, the stirring speed is 140rpm, the residual glucose content in the secondary seed liquid is reduced to 0.5g/L after 18 hours of culture, the shape of the thalli is uniform, the bacteria is good, and the fermentation tank can be transplanted. The formula of the first-level seeding tank culture medium is as follows: 4g/L of glucose, 0.6g/L of yeast powder, 0.2g/L of peptone, 0.8g/L of corn steep liquor, 1.5g/L of ammonium sulfate, 1.7g/L of monosodium glutamate, 1.8g/L of sodium chloride, 3.0g/L of magnesium sulfate, 0.4g/L of potassium dihydrogen phosphate, 0.4g/L of dipotassium hydrogen phosphate, 0.2g/L of ferric chloride, 6g/L of calcium carbonate, 1.8mg/L of thiamine hydrochloride, 3.8mg/L of nicotinic acid, 0.05mg/L of biotin, 6.65 of pH value and 30min of sterilization at the temperature of 121 +/-0.5 ℃. The formula of the culture medium of the secondary seeding tank is as follows: 12g/L of glucose, 0.5g/L of yeast powder, 1.2g/L of corn steep liquor, 2.2g/L of ammonium sulfate, 2.5g/L of monosodium glutamate, 1.8g/L of sodium chloride, 3.8g/L of magnesium sulfate, 0.4g/L of monopotassium phosphate, 0.4g/L of dipotassium phosphate, 0.3g/L of ferric chloride, 4g/L of calcium carbonate, 2.8mg/L of thiamine hydrochloride, 5.5mg/L of nicotinic acid, 0.18mg/L of biotin, pH6.65, and sterilization is carried out at the temperature of 121 +/-0.5 ℃ for 30 min.
Culturing in a fermentation tank: inoculating the cultured seed liquid of the second-stage seed tank into a fermentation tank according to the inoculation amount of 20%, wherein the volume of the liquid in the fermentation tank after inoculation is 40m3The culture conditions of the fermentation tank are as follows: the temperature of the tank is 34 plus or minus 0At 5 ℃, the tank pressure is 0.05 plus or minus 0.005MPa, the aeration ratio is 0.60, and the stirring speed is 120 rpm. The formula of the fermentation tank culture medium is as follows: 13g/L glucose, 2.5g/L corn steep liquor, 2.2g/L ammonium sulfate, 2.8g/L monosodium glutamate, 1.8g/L sodium chloride, 6.5g/L magnesium sulfate, 0.3g/L potassium dihydrogen phosphate, 0.3g/L dipotassium hydrogen phosphate, 0.9g/L ferric chloride, 1.5g/L calcium carbonate, 4.5mg/L thiamine hydrochloride, 8.5mg/L nicotinic acid and 0.20mg/L biotin; lysine hydrochloride 0.03g/L, pH6.45, 121 + -0.5 deg.C sterilization for 30 min.
Fermentation tank process control measures: continuously supplementing 54% glucose solution in the fermentation tank during the culture process, controlling the residual glucose amount in the fermentation tank at 10 + -1 g/L, and stopping supplementing glucose 2 hr before tank placement; continuously supplementing 12% potassium dihydrogen phosphate solution during the culture process, and controlling the phosphorus residue in the fermentation tank at 0.2 + -0.05 g/L; ammonia water with a concentration of 18% is continuously added in the culture process, and the pH value of the fermentation tank is controlled to be 6.7 +/-0.1. Ammonium sulfate solution with the concentration of 30% is continuously supplemented in the fermentation process, and the concentration of ammonium ions in the fermentation tank is controlled as follows: the fermentation period is 1-20h, and the concentration of ammonium ions is 1.2 +/-0.05 g/L; the fermentation period is 21-40h, and the concentration of ammonium ions is 1.5 +/-0.05 g/L; the fermentation period is 41-65h, and the concentration of ammonium ions is 1.9 +/-0.05 g/L; the fermentation period is 66-80h, and the concentration of ammonium ions is 1.4 +/-0.05 g/L; the fermentation period is 81 h-the tank is placed, and the concentration of ammonium ions is 0.9 +/-0.05 g/L.
In the fermentation process, a lysine salt solution with the concentration of 6% is supplemented for 3 times, and the supplementing mode is as follows: supplementing lysine hydrochloride with final concentration of 0.03g/L at 24h of fermentation period; supplementing lysine hydrochloride with final concentration of 0.06g/L at 42h of fermentation period; supplementing lysine hydrochloride with final concentration of 0.01g/L in a fermentation period of 65; adding a sodium sulfate solution with the concentration of 30% in the fermentation process, wherein the adding mode is as follows: sodium sulfate with a final concentration of 0.2g/L is supplemented during the fermentation period of 58 h.
After the cells are cultured for 95 hours in the fermentation tank, the glucose consumption rate of the cells is obviously reduced, the cells are partially autolyzed, the culture in the fermentation tank is stopped and put in the tank, the titer of Q10 put in the tank is 4120mg/L, and the dry weight of the cells is 105 g/L.
Comparative example 1
Seed culture: diluting coenzyme Q10 production strains in a glycerinum tube to a proper concentration, inoculating the strains to a slant culture medium of an eggplant bottle, and culturing the slant in an incubator at the temperature of 32 +/-0.5 ℃ and the relative humidity of 40-60% for 6 days. After the slant culture of the eggplant bottle is finished, 3 colonies are picked by an inoculating needle and inoculated in a 500mL triangular flask filled with 60mL culture medium, and a mother bottle is placed in a shaking table with the temperature of 32 +/-0.5 ℃, the relative humidity of 40-60% and the rotating speed of 220rpm for culture for 30 hours and then used as a mother bottle seed to inoculate a primary seed tank. The eggplant bottle slant culture medium comprises the following components: 2g/L glucose, 1.5g/L yeast powder, 0.5g/L peptone, 1.5g/L sodium chloride, 15g/L agar powder, pH6.75, and sterilizing at 121 + -0.5 deg.C for 20 min. The mother flask culture medium formula is as follows: 5g/L glucose, 0.4g/L yeast powder, 0.3g/L peptone, 0.7g/L corn steep liquor, 1.2g/L ammonium sulfate, 1.5g/L monosodium glutamate, 1.5g/L sodium chloride, 2.5g/L magnesium sulfate, 0.4g/L potassium dihydrogen phosphate, 0.4g/L dipotassium hydrogen phosphate, 0.2g/L ferric chloride, 5.0g/L calcium carbonate, 2.0mg/L thiamine hydrochloride, 4.0mg/L nicotinic acid, 0.1mg/L biotin, pH6.85, and sterilization at 121 +/-0.5 ℃ for 20 min.
First-stage seeding tank expanded culture: inoculating the cultured mother bottle seeds into a first-stage seed tank according to the inoculation amount of 2 thousandth, wherein the volume of a culture medium of the first-stage seed tank is 800L, and the culture conditions of the first-stage seed tank are as follows: the temperature of the tank is 32 plus or minus 0.5 ℃, the pressure of the tank is 0.04 plus or minus 0.005MPa, the aeration ratio is 0.5, the stirring speed is 160rpm, after culturing for 31h, the residual glucose content in the seed liquid is reduced to 0.4g/L, the thallus shape is uniform, the sterility is good, namely, the second-stage seed tank is inoculated according to 10 percent of inoculation amount, the volume of the feed liquid in the second-stage seed tank after inoculation is 8m3The culture conditions of the secondary seeding tank are as follows: the temperature of the tank is 32 plus or minus 0.5 ℃, the pressure of the tank is 0.04 plus or minus 0.005MPa, the aeration ratio is 0.55, the stirring speed is 140rpm, the residual glucose content in the secondary seed liquid is reduced to 0.6g/L after 22 hours of culture, the shape of the thalli is uniform, the bacteria is good, and the fermentation tank can be transplanted. The formula of the first-level seeding tank culture medium is as follows: 5g/L glucose, 0.5g/L yeast powder, 0.2g/L peptone, 0.8g/L corn steep liquor, 1.3g/L ammonium sulfate, 1.5g/L monosodium glutamate, 1.8g/L sodium chloride, 3.0g/L magnesium sulfate, 0.4g/L potassium dihydrogen phosphate, 0.4g/L dipotassium hydrogen phosphate, 0.2g/L ferric chloride, 6g/L calcium carbonate, 1.5mg/L thiamine hydrochloride, 3.5mg/L nicotinic acid, 0.05mg/L biotin, 6.6 pH6, and sterilization is carried out at 121 +/-0.5 ℃ for 30 min. The formula of the culture medium of the secondary seeding tank is as follows: 10g/L of glucose, 0.4g/L of yeast powder, 1.2g/L of corn steep liquor, 2.0g/L of ammonium sulfate and 2.5g/L of monosodium glutamateg/L, sodium chloride 1.8g/L, magnesium sulfate 3.5g/L, potassium dihydrogen phosphate 0.4g/L, dipotassium hydrogen phosphate 0.4g/L, ferric chloride 0.3g/L, calcium carbonate 4g/L, thiamine hydrochloride 2.5mg/L, nicotinic acid 5.0mg/L, biotin 0.15mg/L, pH6.7, sterilization at 121 +/-0.5 ℃ for 30 min.
Culturing in a fermentation tank: inoculating the cultured seed liquid of the second-stage seed tank into a fermentation tank according to the inoculation amount of 20%, wherein the volume of the liquid in the fermentation tank after inoculation is 40m3The culture conditions of the fermentation tank are as follows: the tank temperature is 33 plus or minus 0.5 ℃, the tank pressure is 0.05 plus or minus 0.005MPa, the aeration ratio is 0.65, and the stirring speed is 120 rpm. The formula of the fermentation tank culture medium is as follows: 12g/L glucose, 2.0g/L corn steep liquor, 2.5g/L ammonium sulfate, 2.5g/L monosodium glutamate, 2.0g/L sodium chloride, 5.5g/L magnesium sulfate, 0.3g/L potassium dihydrogen phosphate, 0.3g/L dipotassium hydrogen phosphate, 0.8g/L ferric chloride, 1.5g/L calcium carbonate, 4.0mg/L thiamine hydrochloride, 8.0mg/L nicotinic acid and 0.25mg/L biotin; sterilizing at pH6.45 and 121 + -0.5 deg.C for 30 min.
Fermentation tank process control measures: continuously supplementing 56% glucose solution in the fermentation tank during the culture process, controlling the residual glucose amount in the fermentation tank at 10 + -1 g/L, and stopping supplementing glucose 2 hr before tank placement; continuously supplementing 16% potassium dihydrogen phosphate solution during the culture process, and controlling the phosphorus residue in the fermentation tank at 0.2 + -0.05 g/L; ammonia water with a concentration of 19% is continuously added in the culture process, and the pH value of the fermentation tank is controlled to be 6.7 +/-0.1.
After 85 hours of fermentation tank culture, the glucose consumption rate of thalli is obviously reduced, the thalli is partially autolyzed, the fermentation tank culture is stopped and put into a tank, the Q10 put-into-tank titer is 3075mg/L, and the dry weight of the thalli is 88 g/L.
In comparative example 1, the conditions of the other strains, the formula of the culture medium, the equipment and the like were the same except that the fermentation process control measures proposed by the present invention were not adopted. The results show that the coenzyme Q10 in the comparative example 1 has obviously lower tank-placing titer, dry cell weight and fermentation period, and the batch yield of the coenzyme Q10 is lower. The fermentation process control measures provided by the invention are adopted in the examples 1 and 2, the tank release titer, the thallus dry weight and the fermentation period of the coenzyme Q10 are obviously higher than those of the coenzyme Q10 in the comparative example 1, the batch yield of the coenzyme Q10 is greatly increased, the fermentation cost is obviously reduced, and the brought technical effect is obvious.

Claims (6)

1. A method for fermenting coenzyme Q10 is characterized by comprising the following steps:
1) mother bottle culture: diluting coenzyme Q10 production strains to 1200 strains/mL, inoculating 0.5mL to a slant culture medium, placing the slant culture medium in an incubator at the temperature of 30-34 ℃ and the relative humidity of 40-60% for culturing for 4-8 days, selecting 2-5 colonies, inoculating the colonies in a mother bottle culture medium, placing the mother bottle culture medium in a shaker at the temperature of 30-34 ℃, the relative humidity of 40-60% and the rotation speed of 200-250rpm for culturing for 24-32 hours to obtain mother bottle strains;
2) first-level seeding tank culture: inoculating the strain in the mother bottle to a culture medium in a first-stage seed tank according to the inoculation amount of 1-5 per mill, culturing for 28-36h at 30-35 ℃, the tank pressure of 0.03-0.05MPa, the aeration ratio of 0.3-0.6 and the stirring rotation speed of 140 plus materials of 180rpm to obtain a first-stage seed solution with the glucose residual amount of 0.5-1.0g/L, uniform thallus shape and good sterility;
3) secondary seeding tank culture: inoculating the first-stage seed liquid to a culture medium in a second-stage seed tank according to the inoculation amount of 8-15%, culturing for 16-24h at 30-35 ℃, the tank pressure of 0.03-0.05MPa, the aeration ratio of 0.4-0.7 and the stirring speed of 120-160rpm to obtain the second-stage seed liquid with the glucose residual amount of 0.4-0.8g/L, uniform thallus shape and good sterility;
4) culturing in a fermentation tank: inoculating the second-stage seed liquid into a culture medium in a fermentation tank according to the inoculation amount of 15-25%, culturing at 32-35 ℃, the tank pressure of 0.03-0.06MPa, the aeration ratio of 0.6-0.8 and the stirring speed of 110-140rpm until the glucose consumption rate of thalli is obviously reduced, and stopping fermentation when thalli are partially autolyzed;
continuously supplementing 50-60% glucose solution in the fermentation tank during the culture process to control the residual glucose amount in the fermentation tank at 8-15 g/L; continuously supplementing 13-16% potassium dihydrogen phosphate solution during the culture process to control the phosphorus residue in the fermentation tank at 0.15-0.25 g/L; continuously supplying ammonia water with concentration of 18-22% during the culture process to control pH value in the fermentation tank at 6.5-7.0;
the operation of the fermentation tank culture also comprises the operation of supplementing ammonium salt aqueous solution; the ammonium salt aqueous solution is an aqueous solution of ammonium sulfate, ammonium chloride, ammonium carbonate or ammonium bicarbonate, and the concentration of the ammonium salt aqueous solution is 5-80%; the replenishing mode of the ammonium salt aqueous solution is as follows: when the fermentation period is 1-20h, the ammonium ion concentration is supplemented to the fermentation tank to be 0.8-1.2 g/L; when the fermentation period is 21-40h, the ammonium ion concentration is 1.2-1.6 g/L; when the fermentation period is 41-65h, the ammonium ion concentration is supplemented to the fermentation tank to be 1.6-2.0 g/L; when the fermentation period is 66-80h, the ammonium ion concentration is 1.2-1.6 g/L; the fermentation period is 81 h-when the fermentation tank is placed, the ammonium ion concentration is supplemented to the fermentation tank to be 0.6-1.2 g/L;
the operation of the fermentation tank culture also comprises the operation of supplementing lysine salt aqueous solution; the lysine salt aqueous solution is an aqueous solution of lysine hydrochloride, lysine sulfate or lysine phosphate, and the concentration of the lysine salt aqueous solution is 1-30%; the lysine salt aqueous solution is supplemented in the following mode: when the fermentation period is 20-28h, the lysine salt concentration in the fermentation tank is 0.01-0.1 g/L; when the fermentation period is 40-48h, the lysine salt concentration in the fermentation tank is 0.04-0.12 g/L; when the fermentation period is 60-68h, the lysine salt concentration in the fermentation tank is 0.005-0.05 g/L;
the operation of the fermentation tank culture also comprises the operation of supplementing sulfate aqueous solution; the sulfate aqueous solution is sodium sulfate or potassium sulfate aqueous solution, and the concentration of the sulfate aqueous solution is 10% -60%; the supplementing mode of the sulfate aqueous solution is as follows: when the fermentation period is 36-70h, the lysine salt sulfate concentration in the fermentation tank is 0.2-0.8 g/L.
2. The method of fermenting coenzyme Q10 according to claim 1, wherein the slant medium formulation is: 1.0-4.0g/L of glucose, 0.5-2.0g/L of yeast powder, 0.2-1.0g/L of peptone, 0.5-2.0g/L of sodium chloride, 15-25g/L of agar powder, pH6.8-7.2 and sterilization at 121 ℃ for 20-25 min.
3. The fermentation method of coenzyme Q10 according to claim 1, wherein the mother flask culture medium formula is: 3.0-6.0g/L of glucose, 0.2-0.5g/L of yeast powder, 0.1-0.5g/L of peptone, 0.5-1.0g/L of corn steep liquor, 0.5-2.0g/L of ammonium sulfate, 0.4-2g/L of monosodium glutamate, 0.5-2.0g/L of sodium chloride, 1.0-3.0g/L of magnesium sulfate, 0.1-0.5g/L of potassium dihydrogen phosphate, 0.1-0.5g/L of dipotassium hydrogen phosphate, 0.1-0.5g/L of ferric chloride, 2.0-8.0g/L of calcium carbonate, 1.0-3.0mg/L of thiamine hydrochloride, 2-5mg/L of nicotinic acid, 0.05-0.2mg/L of biotin, 6.5-7.0 of pH value, and 20-25min of sterilization at 121 ℃.
4. The method of claim 1, wherein the medium in the primary seed tank is formulated as: glucose 4.0-8.0g/L, yeast powder 0.3-0.6g/L, peptone 0.1-0.4g/L, corn steep liquor 0.6-1.2g/L, ammonium sulfate 0.8-2.5g/L, monosodium glutamate 0.4-2g/L, sodium chloride 0.5-2.0g/L, magnesium sulfate 1.5-4.0g/L, potassium dihydrogen phosphate 0.1-0.5g/L, dipotassium hydrogen phosphate 0.1-0.5g/L, ferric chloride 0.1-0.5g/L, calcium carbonate 2.0-8.0g/L, thiamine hydrochloride 1.0-3.0mg/L, nicotinic acid 2.0-5.0mg/L, biotin 0.05-0.2mg/L, pH6.5-7.0, and sterilizing at 121 ℃ for 25-30 min.
5. The fermentation method of coenzyme Q10 according to claim 1, wherein the culture medium formulation in the secondary seed tank is: 6.0-12g/L glucose, 0.3-0.6g/L yeast powder, 0.8-1.5g/L corn steep liquor, 1.0-3.5g/L ammonium sulfate, 0.8-3g/L monosodium glutamate, 0.5-2.0g/L sodium chloride, 1.5-4.0g/L magnesium sulfate, 0.1-0.5g/L potassium dihydrogen phosphate, 0.1-0.5g/L dipotassium hydrogen phosphate, 0.1-0.5g/L ferric chloride, 2.0-8.0g/L calcium carbonate, 1.0-3.0mg/L thiamine hydrochloride, 4.0-8.0mg/L nicotinic acid, 0.08-0.3mg/L biotin, 6.5-7.0 pH, and 25-30min sterilization at 121 ℃.
6. The fermentation method of coenzyme Q10 according to claim 1, characterized in that the culture medium formulation in the fermenter is: 10-15g/L of glucose, 1.5-2.5g/L of corn steep liquor, 1.5-3.5g/L of ammonium sulfate, 1.0-3g/L of monosodium glutamate, 0.5-2.0g/L of sodium chloride, 3.0-6.0g/L of magnesium sulfate, 0.1-0.5g/L of potassium dihydrogen phosphate, 0.1-0.5g/L of dipotassium hydrogen phosphate, 0.2-1.0g/L of ferric chloride, 1.0-3.0g/L of calcium carbonate, 2.0-5.0mg/L of thiamine hydrochloride, 6.0-12mg/L of nicotinic acid and 0.1-0.5mg/L of biotin; lysine salt 0.001-5 g/L.
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102168115A (en) * 2010-12-23 2011-08-31 内蒙古金达威药业有限公司 Industrialized production method of coenzyme Q10
CN104561154A (en) * 2014-12-30 2015-04-29 内蒙古金达威药业有限公司 Coenzyme Q10 fermentation process and control strategy
CN105483171A (en) * 2015-12-31 2016-04-13 上虞新和成生物化工有限公司 Production method for increasing industrial output of coenzyme Q10

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102168115A (en) * 2010-12-23 2011-08-31 内蒙古金达威药业有限公司 Industrialized production method of coenzyme Q10
CN104561154A (en) * 2014-12-30 2015-04-29 内蒙古金达威药业有限公司 Coenzyme Q10 fermentation process and control strategy
CN105483171A (en) * 2015-12-31 2016-04-13 上虞新和成生物化工有限公司 Production method for increasing industrial output of coenzyme Q10

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Effects of the Amino Acids of Aspartate Family on the Biosynthesis of CoQ10 in Rhodopseudomonas palustris J001;Jiang Shiyun et al.;《Agricultural Science & Technology》;20081231;第9卷(第4期);摘要，表2 *
产辅酶Q10菌株选育及发酵过程优化硏究;钱大伟;《中国优秀博硕士学位论文全文数据库(硕士)工程科技Ⅰ辑》;20170215(第02期);摘要，第49页第5.2.4节 *

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