CN112553262B - Pravastatin fermentation process - Google Patents
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
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Abstract
The invention belongs to the technical field of fermentation, and particularly discloses a fermentation process of pravastatin, which comprises two steps of seed culture and fermentation culture, wherein the components of a seed culture medium and a fermentation culture medium comprise glucose, yeast extract, soybean peptone and K 2 HPO 4 ·3H 2 O、MgSO 4 ·7H 2 O, trace element solution and defoaming agent. By optimizing the existing fermentation process, the hyphae can keep normal metabolic operation, the speed of the hyphae entering the decline period is delayed, the tendency of the hyphae towards the growth in the fermentation process is corrected, the product inhibition effect is removed, the hyphae can keep higher compactin conversion speed, the biotransformation period is prolonged, the fermentation unit of the product pravastatin is improved, the yield of the pravastatin is finally improved, the aim of efficiently and stably producing the pravastatin is fulfilled, and meanwhile, the raw materials are low in price and beneficial to large-scale industrial production.
Description
Technical Field
The invention belongs to the technical field of fermentation engineering, and particularly relates to a fermentation process of pravastatin.
Background
Coronary heart disease (coronary heart disease) is the most common cardiovascular disease in clinic at present, and the pathological essence of the coronary heart disease is that atherosclerosis lesion occurs in coronary artery, so that blood vessel stenosis or blockage is caused, and further insufficient blood supply and oxygen supply and necrosis of cardiac muscle are caused. Because the lipid in the blood of most coronary heart disease patients is deposited and increased on the vascular wall, atherosclerosis is easy to develop for a long time, and the coronary heart disease patients are easy to block the coronary vessel cavity to cause stenosis, influence the blood flow and form thrombus. Therefore, patients with coronary heart disease need to be treated by antiplatelet and lipid-lowering drugs in clinical treatment. Common antiplatelet drugs include clopidogrel, aspirin, and the like, and both clopidogrel and aspirin have the effect of inhibiting platelet aggregation. The lipid-lowering drug therapy is a basic drug for coronary heart disease, and recent clinical research shows that statins not only have good lipid-regulating effect, but also have good clinical effects in the aspects of anti-inflammation, antioxidation and the like, and the effect of statins in the treatment of cardiovascular and cerebrovascular diseases is concerned and accepted by more and more clinicians.
Pravastatin Sodium (Pravastatin Sodium) is a 3-hydroxy 3-methylglutaryl coenzyme A reductase (HMG-COA reductase) inhibitor, belongs to statins, can effectively reduce serum total cholesterol, low-density lipoprotein and triglyceride and increase high-density lipoprotein by inhibiting the synthesis of cholesterol in a human body, is initially used for treating hyperlipidemia and familial hypercholesterolemia, can slow down the development of atherosclerosis, reduce the occurrence of coronary atherosclerotic lesions and clinical cardiovascular events, improve the cardiac function and improve the left ventricular ejection fraction, and has very important effects on the aspects of retroversion left ventricle reconstruction, prevention and treatment of myocardial fibrosis and the like, and the application of the Pravastatin Sodium is a drug for treating the cardiovascular diseases; after the pravastatin is taken for a long time, the death rate caused by various reasons can be reduced no matter whether a patient has coronary heart disease or not, and the pravastatin becomes the only medicine which can be used for the first-level and second-level defense of the heart disease and the stroke of the patient with higher cholesterol level or coronary heart disease in the prior statin drugs. According to the existing research, pravastatin not only has the functions of lipid regulation and anti-inflammation, but also can reduce the incidence rate of diabetes and Alzheimer disease (senile dementia); can inhibit proliferation of hepatocarcinoma cell; can effectively treat ulcerative colitis, has little side effect and low half-year recurrence rate; can improve cardiac function and improve ventricular remodeling in the aspect of treating chronic heart failure and diastolic heart failure.
Production of pravastatin sodium is generally obtained by a two-step fermentation process, and reports or patents (WO 99/10499, WO2007/147827, US 6,274,360 and EP 1,266,967) describe that pravastatin can be produced by a one-step fermentation method, but due to technical complexity, production efficiency and the like, industrial production of pravastatin is mainly performed by two-step fermentation. The two-step fermentation process comprises the following steps: first, a secondary metabolite Compactin (also known as Mevastatin) is obtained by microbial fermentation of Penicillium citrinum or the like, and then, by the conversion of microbial enzymes, the C-6 carbonyl group on the hexahydronaphthalene ring of a Compactin salt (usually an alkali metal or ammonium salt) is hydroxylated to form pravastatin sodium. For example, patent document CN102757986a discloses a fermentation process for producing pravastatin by converting compactin with actinomadura, wherein a trace element solution is added into a seed culture medium to improve the conversion capability of actinomadura to compactin, so as to improve the yield of pravastatin; patent document CN108070623A discloses a method for improving pravastatin fermentation yield, which effectively improves the tolerance of actinomadura bacteria to mevastatin by adding bran extract in a fermentation medium, so as to improve the pravastatin yield.
The existing pravastatin fermentation process generally has the following defects: carbon sources and nitrogen sources are all used as basic materials to be put into a culture medium, so that the concentrations of the carbon sources and the nitrogen sources are high in the early stage of fermentation, and the growth of hyphae is inhibited; at the later stage, hyphae lack of sufficient metabolic energy due to too low concentrations of the carbon source and the nitrogen source, so that the decay speed of the hyphae is higher, and the fermentation yield of pravastatin cannot be further improved. In addition, due to the product inhibition or strain growth characteristics, hypha growth is biased in the middle stage of fermentation, and the capacity of converting compactin into pravastatin is inhibited, so that more pravastatin products cannot be obtained. Therefore, it is especially important to develop a pravastatin production process that can remove product inhibition, improve the fermentation unit of the product pravastatin, and simultaneously can produce pravastatin efficiently and stably.
Disclosure of Invention
The invention aims to provide a fermentation process of pravastatin, and particularly relates to a process for producing pravastatin by converting compactin with actinomadura actinomycetes, which has the functions of correcting the tendency of fermentation to hypha growth, removing product inhibition and promoting microbial growth, so that hypha keeps higher conversion rate of the compactin, prolongs the biotransformation period, and finally improves the fermentation unit of the product pravastatin.
In order to achieve the purpose, the invention adopts the following technical scheme: a fermentation process of pravastatin comprises the following steps:
s1, seed culture: the seed culture medium is prepared according to the following formula: 25.0-35.0 g/L of glucose, 15.0-25.0 g/L of yeast extract and 3.0-7.0 g/L, K of soybean peptone 2 HPO 4 ·3H 2 O 1.0～3.0g/L、MgSO 4 ·7H 2 0.1-1.0 g/L of O and 1.0-5.0 ml/L of trace element solution A, and adjusting the pH value to 7.0-7.2; inoculating actinomadura bacteria to a seed culture medium, and culturing at 35-37 ℃ for 48-72 hours to obtain a seed solution;
s2, fermentation culture: the fermentation medium is prepared according to the following formula: 25.0-35.0 g/L of glucose, 15.0-25.0 g/L of yeast extract and 3.0-7.0 g/L, K of soybean peptone 2 HPO 4 ·3H 2 O 1.0～3.0g/L、MgSO 4 ·7H 2 O 0.1～1.0g/L、(NH 4 ) 2 SO 4 1.0-3.0 g/L, 1.0-5.0 ml/L of trace element solution A and 0.5-2.0 g/L of defoaming agent, adjusting the pH value to 7.0-7.2, inoculating the seed solution obtained in the step S1 into a fermentation culture medium, and fermenting and culturing at the temperature of 27-37 ℃ for 7-15 days to obtain fermentation liquor containing pravastatin.
Preferably, the trace element solution A is prepared according to the following formula: sodium molybdate (Na) 2 MoO 4 ·2H 2 0.1-0.2 g/L of O) and boric acid (H) 3 BO 3 ) 1.0-1.5 g/L of manganese chloride (MnCl) 2 ·H 2 0.1-0.2 g/L of O) and zinc sulfate (ZnSO) 4 ·7H 2 O) 5.0-5.5 g/L, ferrous sulfate (FeSO) 4 ·7H 2 O1.5-2.0 g/L, sodium nitrate (NaNO) 3 ) 3.0 to 3.3g/L of citric acid (C) 6 H 8 O 7 ) 1.5-2.0 g/L and copper sulfate (CuSO) 4 ·5H 2 O)0.2～0.3g/L。
Preferably, in the fermentation culture process in the step S2, a compactin sodium solution is supplemented to maintain the concentration of compactin sodium in the fermentation broth to be 0.3-0.7 g/L.
Preferably, in the fermentation culture process of the step S2, the trace element solution B is supplemented when the culture time is up to 100 hours. Specifically, the supplement of a proper trace element solution can correct the tendency of hypha growth in the fermentation process, and can effectively remove the product inhibition effect, so that the hypha can keep a higher compactin conversion speed, the biotransformation period can be prolonged, and the fermentation unit of the product pravastatin can be finally improved.
Preferably, the trace element solution B is prepared according to the following formulaObtaining: sodium molybdate (Na) 2 MoO 4 ·2H 2 0.3-0.4 g/L of O) and boric acid (H) 3 BO 3 ) 1.0-1.5 g/L of manganese chloride (MnCl) 2 ·H 2 0.3-0.4 g/L of O and zinc sulfate (ZnSO) 4 ·7H 2 5.0-5.5 g/L of O) and ferrous sulfate (FeSO) 4 ·7H 2 1.5-2.0 g/L of O) and sodium nitrate (NaNO) 3 ) 3.0 to 3.5g/L of citric acid (C) 6 H 8 O 7 ) 1.5-2.0 g/L of valine (C) 5 H 11 NO 2 ) 1.0-3.0 g/L leucine (C) 6 H 13 NO 2 ) 1.0-3.0 g/L and copper sulfate (CuSO) 4 ·5H 2 O)0.4～0.5g/L。
Preferably, during the fermentation culture process in the step S2, soybean peptone solution with the concentration of 10g/L is supplemented to maintain the total nitrogen concentration during the culture process to be more than 15 mg/L. Specifically, the metabolism is vigorous in the early stage of fermentation, the consumption of the nitrogen source is rapid, and the proper amount of the nitrogen source is supplemented in order to keep the nitrogen source sufficient in the fermentation process, so that the normal metabolism can be ensured before the mycelium enters the stabilization stage.
Preferably, in the fermentation culture process of step S2, sucrose solution with a concentration of 30-80% is supplemented to maintain the total sugar concentration above 25g/L in the culture process. Specifically, when the sugar concentration in the middle and later stages of fermentation is low, the sucrose solution is timely supplemented to maintain the total sugar concentration of the fermentation liquor, so that the speed of hyphae entering the decline period can be reduced.
Preferably, the antifoaming agent is polyoxyethylene polyoxypropylene glyceryl ether.
Therefore, compared with the prior art, the invention has the following beneficial effects:
(1) The invention adds soybean peptone solution, sucrose solution and microelement solution with different formula from the culture medium in the middle fermentation period of the existing pravastatin fermentation technology, can keep the normal operation of hypha metabolism, delay the speed of hypha entering the death period, remove the inhibition of the compactin product, keep the hypha with higher conversion speed of the compactin, prolong the biotransformation period and finally improve the yield of the product pravastatin.
(2) The fermentation process has the characteristics of high fermentation level and stable fermentation process.
(3) The process has low production cost and is suitable for large-scale industrial production.
Detailed Description
The present invention will be further described below by way of specific embodiments, but the present invention is not limited to only the following examples. It will be apparent to those skilled in the art that the invention can be modified and replaced with other components having the same effects without departing from the spirit and scope of the invention, and all such modifications and substitutions are deemed to be within the scope of the invention.
The experimental procedures, in which specific conditions are not specified, in the following examples are generally conducted under conventional conditions or conditions recommended by manufacturers.
In the following examples of the present invention Actinomadura maduram belongs to the same genus as that of American Type Culture Collection (ATCC) No. 45678 (WO 96/40863).
In the following examples of the present invention, the contents of pravastatin and compactin were measured according to the method of Han Minghuo, etc. "content of relevant components in pravastatin production sample by HPLC method" pharmaceutical for today 2010, 20 (7).
In the examples of the present invention, the conversion of compactin was calculated as follows:
conversion = pravastatin yield ÷ compactin usage × 100%.
Example 1
This example uses a 5L full-automatic fermentation tank to produce pravastatin
S1, seed culture: the seed culture medium is prepared according to the following formula: 30.0g/L glucose, 20.0g/L yeast extract, 5.0g/L, K soy peptone 2 HPO 4 ·3H 2 O 2.0g/L、MgSO 4 ·7H 2 0.5g/L of O and 2.0ml/L of trace element solution A, and adjusting the pH value to 7.0; inoculating actinomadura bacteria to a seed culture medium, and culturing at 35-37 ℃ for 48 hours to obtain a seed solution;
s2, fermentation culture: the fermentation medium is prepared according to the following formula: 35.0g/L glucose, 25.0g/L yeast extract, 5.0g/L, K soy peptone 2 HPO 4 ·3H 2 O 2.0g/L、MgSO 4 ·7H 2 O 0.5g/L、(NH 4 ) 2 SO 4 2.0g/L, 2.0mL/L of trace element solution A, 1.0g/L of polyoxyethylene polyoxypropylene glycerol ether, adjusting pH to 7.2, inoculating 300mL of seed solution obtained in the step S1 after sterilization of a fermentation medium, performing fermentation culture for 12 days under the conditions of 35-37 ℃, stirring speed of 300rpm, air flow of 0.5vvm and tank pressure of 0.05Mpa, supplementing compactin solution after culturing for 36 hours, detecting the concentration of the compactin by an HPLC method to maintain the concentration of the compactin at 0.3-0.7g/L, and supplementing 10g/L of soybean peptone solution after 80-150 hours of fermentation liquor to maintain the total nitrogen concentration of the culture process at more than 15 mg/L. The fermentation liquor is cultured for 100 hours, and the trace element solution B is added into the fermentation liquor at one time according to the volume ratio of 0.5 ml/L. And (3) supplementing 30-80% of sucrose solution after the fermentation broth is used for 150 hours, maintaining the total sugar concentration in the culture process at 25g/L, culturing until the consumption rate of compactin is less than 100mg/hr, stopping supplementing the compactin solution, and detecting the content of pravastatin in the fermentation broth after the fermentation is finished.
The trace element solution A is prepared according to the following formula (g/L):
sodium molybdate (Na) 2 MoO 4 ·2H 2 O) 0.1g/L, boric acid (H) 3 BO 3 ) 1.5g/L, manganese chloride (MnCl) 2 ·H 2 O) 0.1g/L, zinc sulfate (ZnSO) 4 ·7H 2 O) 5.0g/L, ferrous sulfate (FeSO) 4 ·7H 2 O) 2.0g/L, sodium nitrate (NaNO) 3 ) 3.0g/L of citric acid (C) 6 H 8 O 7 ) 1.5g/L and copper sulfate (CuSO) 4 ·5H 2 O)0.2g/L。
The trace element solution B is prepared according to the following formula (g/L):
sodium molybdate (Na) 2 MoO 4 ·2H 2 O) 0.3g/L, boric acid (H) 3 BO 3 ) 1.5g/L, manganese chloride (MnCl) 2 ·H 2 O) 0.3g/L, zinc sulfate (ZnSO) 4 ·7H 2 O) 5.5g/L, ferrous sulfate (FeSO) 4 ·7H 2 O) 2.0g/L, sodium nitrate (NaNO) 3 ) 3.0g/L of citric acid (C) 6 H 8 O 7 ) 1.5g/L, valine (C) 5 H 11 NO 2 ) 1.0g/L leucine (C) 6 H 13 NO 2 )1.0g/LAnd copper sulfate (CuSO 4 & 5H) 2 O)0.4g/L。
Example 2
This example uses a 50L full-automatic fermentation tank to produce pravastatin
S1, seed culture: the seed culture medium is prepared according to the following formula: 30.0g/L glucose, 20.0g/L yeast extract, 5.0g/L, K soy peptone 2 HPO 4 ·3H 2 O 2.0g/L、MgSO 4 ·7H 2 0.5g/L of O and 2.0ml/L of trace element solution A, and adjusting the pH value to 7.0; inoculating actinomadura in a seed culture medium, and culturing for 72 hours at 35-37 ℃ to obtain a seed solution;
s2, fermentation culture: A30L fermentation medium was prepared according to the following formulation: 35.0g/L glucose, 25.0g/L yeast extract, 5.0g/L soyabean peptone, K 2 HPO 4 ·3H 2 O 2.0g/L，MgSO 4 ·7H 2 O 0.5g/L，(NH 4 ) 2 SO 4 2.0g/L, 2.0ml/L of trace element solution A and 1.0g/L of polyoxyethylene polyoxypropylene glycerol ether, and adjusting the pH value to 7.2. Inoculating 2.5L of seed liquid obtained in the step S1 after a fermentation culture medium is sterilized, fermenting and culturing for 15 days under the conditions of 35-37 ℃, stirring speed of 300rpm, air flow of 0.5vvm and tank pressure of 0.05Mpa, supplementing a compactin solution after culturing for 36h, detecting the compactin concentration by an HPLC method to maintain the compactin concentration at 0.3-0.7g/L, and supplementing a soybean peptone solution with the concentration of 10g/L into a fermentation liquid for 80-150 h to maintain the total nitrogen concentration in the culture process at more than 15 mg/L. The fermentation liquor is cultured for 100 hours, and the trace element solution B is added into the fermentation liquor at one time according to the volume ratio of 0.5 ml/L. And (3) supplementing 30-80% of sucrose solution after the fermentation broth is used for 150 hours, maintaining the total sugar concentration in the culture process at 25g/L, culturing until the consumption rate of compactin is less than 100mg/hr, stopping supplementing the compactin solution, and detecting the content of pravastatin in the fermentation broth after the fermentation is finished.
The trace element solution A is prepared according to the following formula (g/L):
sodium molybdate (Na) 2 MoO 4 ·2H 2 O) 0.1g/L, boric acid (H) 3 BO 3 ) 1.5g/L, manganese chloride (MnCl) 2 ·H 2 O) 0.1g/L, zinc sulfate (ZnSO) 4 ·7H 2 O) 5.0g/L, ferrous sulfate (FeSO) 4 ·7H 2 O) 2.0g/L, sodium nitrate (NaNO) 3 ) 3.0g/L of citric acid (C) 6 H 8 O 7 ) 1.5g/L and copper sulfate (CuSO) 4 ·5H 2 O)0.2g/L。
The trace element solution B is prepared according to the following formula (g/L):
sodium molybdate (Na) 2 MoO 4 ·2H 2 O) 0.3g/L, boric acid (H) 3 BO 3 ) 1.5g/L, manganese chloride (MnCl) 2 ·H 2 O) 0.3g/L, zinc sulfate (ZnSO) 4 ·7H 2 O) 5.5g/L, ferrous sulfate (FeSO) 4 ·7H 2 O) 2.0g/L, sodium nitrate (NaNO) 3 ) 3.0g/L of citric acid (C) 6 H 8 O 7 ) 1.5g/L, valine (C) 5 H 11 NO 2 ) 1.0g/L leucine (C) 6 H 13 NO 2 ) 1.0g/L and copper sulfate (CuSO4.5H) 2 O)0.4g/L。
Comparative example 1
The comparative example differs from example 2 only in that: the fermentation process does not supplement any fermentation liquor.
Comparative example 2
The comparative example differs from example 2 only in that: the trace element solution B is not supplemented in the fermentation process.
Comparative example 3
The comparative example differs from example 2 only in that: no soybean peptone solution and sucrose solution were supplemented.
Comparative example 4
The comparative example differs from example 2 only in that: the trace element solution formula supplemented in the middle stage of fermentation is not added with leucine and valine, and correspondingly the content of sodium nitrate is increased to 4.0g/L, and the content of copper sulfate is increased to 1.4g/L.
Comparative example 5
The comparative example differs from example 2 only in that: the formula of the microelement solution supplemented in the middle stage of fermentation is not added with leucine, and correspondingly, the content of copper sulfate is increased to 1.4g/L.
Test example I. Compactin conversion test
1. Experimental samples: examples 1-2, comparative examples 1-5 fermentation broths.
2. Results of the experiment
The experimental data of the fermentation broths of examples 1 to 2 and comparative examples 1 to 5 are shown in Table 1.
TABLE 1 results of Pravastatin conversion data
As can be seen from the experimental data in Table 1, the conversion and fermentation levels of compactin are higher in examples 1-2 of the present invention than in comparative examples 1-3. From the experimental data of the examples 1-2 and the comparative examples 1-2, it can be known that the addition of the soybean peptone solution and the sucrose solution in the fermentation process can effectively improve the conversion capability of the fermented mycelia on the compactin, keep the normal operation of the mycelia metabolism, and delay the speed of the fermented mycelia entering the decline period, so as to improve the fermentation unit of the pravastatin.
According to the data of the comparative examples 1 and 3, the trace element solution is supplemented in the fermentation culture period of 100 hours, so that the tendency of hypha growth in the fermentation process can be effectively corrected, the product inhibition effect is removed, the hypha keeps higher compactin conversion speed, the biotransformation period is prolonged, and the fermentation unit of the product pravastatin is finally improved, namely the total yield of pravastatin in the fermentation period is improved.
It can be concluded from the data in examples 1-2 and comparative examples 4-5 that examples 1-2 of the present invention have the effect of significantly enhancing the fermentation unit of pravastatin and prolonging the fermentation period of microorganisms, i.e., significantly increasing the total amount of pravastatin produced during the fermentation period. In comparative examples 4 to 5, due to the lack of valine and leucine components, no synergistic effect between the components could be formed, resulting in a lower pravastatin content in the final fermentation broth than in examples 1 to 2 of the present invention.
On the background that the conversion rate of the existing pravastatin fermentation technology is difficult to improve, a trace element solution, a soybean peptone solution and a sucrose solution with a new formula are added into a fermentation medium for 100 hours, so that product inhibition can be effectively eliminated, hypha deviation to the hypha growth direction is corrected, the intermediate product inhibition effect is removed, the growth of actinomadura bacteria is promoted, the action of the hypha entering the decay period speed is delayed, the hypha keeps higher conversion rate of the compactin, the biotransformation period is prolonged, and the purpose of obviously improving the pravastatin fermentation yield is finally achieved.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.
Claims (2)
1. The fermentation process of pravastatin is characterized by comprising the following steps:
s1, seed culture: the seed culture medium is prepared according to the following formula: 30.0g/L glucose, 20.0g/L yeast extract, 5.0g/L, K soy peptone 2 HPO 4 ·3H 2 O2.0 g/L、MgSO 4 ·7H 2 O0.5 g/L and a trace element solution A2.0 ml/L, and adjusting the pH value to 7.0-7.2; inoculating actinomadura into a seed culture medium, and culturing at 35-37 ℃ for 48-72 hours to obtain a seed solution;
s2, fermentation culture: the fermentation medium is prepared according to the following formula: 35.0g/L glucose, 25.0g/L yeast extract, 5.0g/L, K soytone 2 HPO 4 ·3H 2 O 2.0 g/L、MgSO 4 ·7H 2 O0.5 g/L、(NH 4 ) 2 SO 4 2.0g/L, 2.0ml/L of the trace element solution A and 1.0g/L of an antifoaming agent, adjusting the pH to 7.0-7.2, inoculating the seed solution obtained in the step S1 into a fermentation culture medium, and performing fermentation culture at 35-37 ℃ for 7-15 days to obtain a fermentation liquid containing pravastatin;
in the fermentation process of the step S2, after culturing for 36 hours, supplementing a compactin solution to maintain the concentration of the compactin in the fermentation liquor to be 0.3-0.7g/L, and supplementing a soybean peptone solution with the concentration of 10g/L to maintain the total nitrogen concentration of the fermentation liquor to be more than 15mg/L in 80-150 hours; the fermentation liquor is cultured for 100 hours, and the trace element solution B is supplemented into the fermentation liquor at one time according to 0.5 ml/L; after 150 hours, adding 30-80% of sucrose solution into the fermentation liquor to maintain the total sugar concentration at more than 25g/L in the culture process;
the trace element solution A is prepared according to the following formula: na (Na) 2 MoO 4 ·2H 2 O0.1g/L, boric acid (H) 3 BO 3 )1.5 g/L、MnCl 2 ·H 2 O0.1 g/L、ZnSO 4 ·7H 2 O5.0 g/L、FeSO 4 ·7H 2 O2.0 g/L, sodium nitrate (NaNO) 3 ) 3.0g/L, citric acid (C) 6 H 8 O 7 ) 1.5g/L and CuSO 4 ·5H 2 O0.2 g/L；
The trace element solution B is prepared according to the following formula: na (Na) 2 MoO 4 ·2H 2 O0.3g/L, boric acid (H) 3 BO 3 )1.5 g/L、MnCl 2 ·H 2 O0.3 g/L、ZnSO 4 ·7H 2 O5.5 g/L、FeSO 4 ·7H 2 O2.0 g/L, sodium nitrate (NaNO) 3 ) 3.0g/L, citric acid (C) 6 H 8 O 7 ) 1.5g/L, valine (C) 5 H 11 NO 2 ) 1.0g/L leucine (C) 6 H 13 NO 2 ) 1.0g/L and CuSO 4 ·5H 2 O0.4g/L。
2. The pravastatin fermentation process of claim 1, wherein the antifoaming agent is a polyoxyethylene polyoxypropylene glyceryl ether.
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| CN109913509A (en) * | 2018-04-28 | 2019-06-21 | 湖北大学 | A kind of fermentation medium and zymotechnique of efficient production short-chain branch fatty acid |
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| US6682913B1 (en) * | 1999-02-03 | 2004-01-27 | Institute For Drug Research Ltd. | Microbial process for preparing pravastatin |
| EP2383260A3 (en) * | 2010-04-30 | 2011-12-28 | Dipharma Francis S.r.l. | Process for the preparation of statins |
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