CN114045313A - Method for regulating and controlling citric acid fermentation by controlling aspergillus niger bacteria ball form - Google Patents
Method for regulating and controlling citric acid fermentation by controlling aspergillus niger bacteria ball form Download PDFInfo
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Abstract
The invention relates to the field of citric acid production by Aspergillus niger fermentation, and particularly relates to a method for regulating and controlling citric acid fermentation by controlling the shape of Aspergillus niger bacteria balls. In the process of Aspergillus niger seed culture or fermentation, the Aspergillus niger in the process of seed culture or fermentation is enabled to form a multi-branch shape with compact and regular mycelium, small diameter and short and strong mycelium by reasonably controlling the flow rate of a carbon source or a nitrogen source and limiting the concentrations of phosphorus and manganese elements in a culture medium. The Aspergillus niger shape is beneficial to the transfer of nutrient substances, oxygen and metabolic target product citric acid, accelerates the fermentation to produce acid and shortens the fermentation period; meanwhile, hypha shortening is beneficial to reducing the viscosity of fermentation liquor and reducing the power consumption of stirring; the method is beneficial to filtering the fermentation liquor to form a bridging effect, improves the filtering effect, greatly improves the extraction yield and the filtrate quality, reduces the water content of filter residues, and improves the comprehensive economic benefit of enterprises.
Description
Technical Field
The invention relates to the field of citric acid production by Aspergillus niger fermentation, and particularly relates to a method for regulating and controlling citric acid fermentation by controlling the shape of Aspergillus niger bacteria balls.
Background
The citric acid can be used in beverage, food, and pharmaceutical industry. Its use depends on three characteristics: acidity, taste and salt formation. Citric acid can form a wide range of salts which can be used in industry as masking and anticoagulant protectants, also in fats and oils, to act as an antioxidant; citric acid can also form esters with many alcohols, such as butyl, triethyl and acetyl tributyl esters, which are useful as plastic film plasticizers.
Aspergillus niger is in liquid submerged culture, and exhibits different morphology depending on culture conditions, and the morphology of Aspergillus niger is critical for the fermentation yield of citric acid. At present, the aspergillus niger morphology mainly presents two forms: filamentous bacteria, smooth compact bacteria balls. The filamentous fungi have the main disadvantages that: filamentous aspergillus niger increases the viscosity of fermentation liquor, and the subsequent influence is that the stirring and mass transfer difficulty is increased; the defects of the smooth and compact bacterium ball are as follows: the growth and acid production of cells can only occur on the surface of the compact mycelium pellet, enough nutrients and oxygen can be contacted on the surface, and the production of citric acid is limited by the transmission limitation of nutrients, oxygen and the like of the cells growing in the mycelium pellet. Therefore, the culture of the Aspergillus niger form which is beneficial to the transfer of nutrient substances and oxygen and simultaneously reduces the viscosity of fermentation liquor is important, so that the acid production speed is accelerated, the fermentation period is shortened, and the power consumption is reduced.
Disclosure of Invention
Aiming at the defects of the existing Aspergillus niger culture method, the invention provides a method for controlling the Aspergillus niger morphology to regulate and control citric acid fermentation. The main technical scheme is that in the Aspergillus niger seed culture or fermentation process, the Aspergillus niger in the seed culture or fermentation process forms a compact, regular and small-diameter mycelium with short and strong mycelium and a multi-branch shape by reasonably controlling the flow rate of a carbon source or a nitrogen source and limiting the concentrations of phosphorus element and manganese element in a culture medium. The Aspergillus niger shape is beneficial to the transfer of nutrient substances, oxygen and metabolic target product citric acid, accelerates the fermentation to produce acid and shortens the fermentation period; meanwhile, hypha shortening is beneficial to reducing the viscosity of fermentation liquor and reducing the power consumption of stirring; the method is beneficial to filtering the fermentation liquor to form a bridging effect, improves the filtering effect, greatly improves the extraction yield and the filtrate quality, reduces the water content of filter residues, and improves the comprehensive economic benefit of enterprises.
The invention provides a method for regulating and controlling citric acid fermentation by controlling the shape of an Aspergillus niger strain ball, which comprises 1) Aspergillus niger seed culture and 2) citric acid fermentation process.
Wherein, in the step 1) aspergillus niger seed culture process:
the phosphorus element concentration in the seed basic culture medium is as follows: 2.5-3.5g/L, the concentration of manganese element is as follows: 0.06-0.10 mg/L;
the Aspergillus niger seed culture stage controls the flow rate of a carbon source and a nitrogen source to regulate the Aspergillus niger form, and specifically comprises the following steps: controlling the carbon source flow acceleration rate to be 0.5-0.7g sugar/(L.h) and the nitrogen source flow acceleration rate to be 55-66 mg/(L.h), wherein the ratio of inorganic nitrogen to organic nitrogen is 0.5:1-0.8:1, and the C/N ratio is 3.0:1-5.2: 1; the carbon source flow rate is controlled to be 1.1-1.3g sugar/(L.h) in 9h-16h, and the nitrogen source flow rate is controlled to be 100-120 mg/(L.h), wherein the ratio of inorganic nitrogen to organic nitrogen is 0.4:1-0.65:1, and the C/N is 3.6:1-5.2: 1; 17h-24h, the carbon source flow rate is controlled to be 1.8-2.0g sugar/(L.h), the nitrogen source flow rate is controlled to be 100-120 mg/(L.h), wherein the nitrogen sources are all organic nitrogen sources, and the C/N is 6.0:1-7.8: 1.
The specific process comprises the following steps: inoculating the aspergillus niger spore suspension into a sterilized seed basal culture medium in an aseptic mode, and controlling seed culture conditions to culture aspergillus niger seeds, wherein the seed culture conditions are as follows: the pressure in the tank is 0.06-0.12Mpa, the temperature in the tank is 32-40 ℃, the ventilation ratio is 0.15-0.23vvm, and the stirring speed is 250-350 rpm.
Preferably, in the step 1) Aspergillus niger seed culture stage, the Aspergillus niger spore inoculation concentration is 25-50 ten thousand/mL.
Preferably, in the step 1) aspergillus niger seed culture process, the seed basal medium comprises the following components:
(1) nutrient salt: KCl 4-6g/L, phosphorus-containing nutrient salt (phosphorus concentration: 2.5-3.5g/L), MgSO4·7H2O 0.25-0.4g/L、Na2SO42g/L of anhydrous CaCl2 4-6g/L；
(2) Trace elements: FeSO4·7H2O 1.5-2.5mg/L、ZnSO4·7H20.4-0.6mg/L of O, manganese-containing nutrient salt (the concentration of manganese element is controlled to be 0.06-0.10mg/L), and CuSO4·5H2O 0.15-0.25mg/L、CoCl2·6H2O0.10-0.20 mg/L; and (5) sterilizing for later use.
2) In the citric acid fermentation process:
in the fermentation basic culture medium, the concentration of phosphorus element is as follows: 0.55-0.80g/L, manganese element concentration is as follows: 0.04-0.06 mg/L;
controlling the feeding speed of the nitrogen source according to different fermentation stages to regulate the shape of the Aspergillus niger: controlling the nitrogen source flow acceleration rate to be 15-17 mg/(L.h) for 0h-20h, wherein the nitrogen source is an organic nitrogen source; and (3) 21 h-ending fermentation, wherein the feeding acceleration of a nitrogen source is controlled to be 13-15 mg/(L.h), and the nitrogen source is an organic nitrogen source.
The specific process comprises the following steps: after the Aspergillus niger seed liquid is cultured, transferring the seed liquid into a sterilized and cooled fermentation basal culture medium, and controlling fermentation conditions to start fermentation; the fermentation culture conditions are as follows: the pressure in the tank is 0.06-0.12Mpa, the temperature in the tank is 35-40 ℃, the ventilation ratio is 0.18-0.25vvm, and the stirring speed is 300-400 rpm.
Preferably, the inoculation concentration of the bacteria balls in the fermentation stage is controlled to be 2.5-5.0 ten thousand/mL.
Preferably, the fermentation base medium consists of:
(1) carbon source: total sugar (175-;
(2) nutrient salt: KCl 2.5-4.0g/L, phosphorus-containing nutrient salt (the concentration of phosphorus element is controlled to be 0.55-0.80g/L), MgSO4·7H2O 0.15-0.35g/L、Na2SO40.40-0.55g/L of anhydrous CaCl2 6.0-8.0g/L；
(3) Trace elements: FeSO4·7H2O 4.0-6.0mg/L、ZnSO4·7H2O2.0-3.0 mg/L, manganese-containing nutrient salt (the concentration of manganese element is controlled to be 0.04-0.06mg/L), CuSO4·5H2O 0.08-0.12mg/L、CoCl2·6H2O0.07-0.11 mg/L, biotin 2.5-4.0mg/L, Vb 11.8-2.5 mg/L;
1) in the processes of Aspergillus niger seed culture and 2) citric acid fermentation, the carbon source refers to one or more of starch (corn, potato, cassava), glucose and the like; the inorganic nitrogen refers to one or more of ammonia water, ammonium salt or nitrate (ammonium sulfate, ammonium nitrate, sodium nitrate, etc.), urea, etc., and the organic nitrogen refers to one or more of corn steep liquor, bean cake powder, peanut powder, yeast extract, etc.; the phosphorus-containing nutrient salt refers to one or more of potassium dihydrogen phosphate, dipotassium hydrogen phosphate and the like; the manganese-containing nutritive salt refers to one or more of manganese sulfate, manganese chloride and the like.
In the application, the growth of the aspergillus niger is limited by controlling the concentrations of a carbon source and a nitrogen source (the concentration unit of the sugar in the measured concentration: sugar/(L.h) refers to glucose, and the concentration of the carbon source is measured according to the glucose) in a culture medium in the aspergillus niger seed culture stage, so that the specific growth rate of the aspergillus niger is maintained in a proper range, and the hypha branching frequency is reduced, so that the aspergillus niger forms a shape with compact, regular and small hyphae, short and thick hyphae and more branching degree. However, the lack of the nitrogen source can affect the growth of aspergillus niger, and the increase of the concentration of the nitrogen source can cause the formation of larger and firmer bacteria balls, which is not beneficial to the transfer of nutrient substances and oxygen, and simultaneously consumes additional carbon source and affects the conversion rate of saccharic acid.
In the aspergillus niger seed culture stage, the initial concentration of phosphorus element is controlled as follows: 2.5-3.5g/L, and controlling the initial concentration of manganese element as follows: 0.06-0.10 mg/L. Even if a nitrogen source and a carbon source are added during the culture process, the concentrations of phosphorus and manganese elements are still in a proper range, and the concentrations of the two elements play an important role in the hypha morphology. At higher phosphorus content, the mycelium loosens, resulting in increased viscosity, reduced dissolved oxygen, decreased yield, and lower phosphorus content affecting thallus growth. Manganese ions participate in processes such as cell wall synthesis and spore formation, and lack of manganese ions can cause abnormal aspergillus niger morphology, spore expansion, formation of nodular hyphae, glycoprotein turnover inhibition and reduction of polar growth of hyphae. Too high manganese ions can change the shape of the aspergillus niger, the mycelium does not agglomerate, and the diameter of the mycelium is reduced.
On the basis that aspergillus niger is enabled to obtain corresponding forms by the aspergillus niger seed culture, the initial concentration of phosphorus is controlled to be as follows in the citric acid fermentation process: 0.55-0.80g/L, and the initial concentration of manganese element is as follows: 0.04-0.06mg/L, and further controlling the fermentation state of the Aspergillus niger by feeding a nitrogen source and a carbon source.
Compared with the prior art, the method is mainly characterized in that the method for controlling the feeding speed of the glucose and the nitrogen source, limiting the concentrations of the phosphorus element and the manganese element and the like in the aspergillus niger seed culture stage enables aspergillus niger to form a shape with compact, regular and small mycelium, short and strong mycelium and more branching degrees, and the aspergillus niger shape is beneficial to fermentation for producing citric acid and subsequent extraction processes. In conclusion, the aspergillus niger cultured by the method has the advantages that the acid production speed is accelerated, the sugar-acid conversion rate is improved, and the fermentation period is shortened; meanwhile, the stirring power consumption is reduced.
Drawings
FIG. 1A. niger bacterial pellet in example 1 is a 100-fold and 400-fold microscopic view respectively;
FIG. 2. example 2A. niger mycosphere under 100-fold and 400-fold microscope respectively;
FIG. 3A. niger bacterial pellet in example 3 under 100-fold and 400-fold microscope view field respectively;
FIG. 4 image of field of view of Aspergillus niger bacterial pellet under 100-fold and 400-fold microscope respectively;
FIG. 5A. niger bacterial pellet under 100-fold and 400-fold microscope view field image respectively in example 5;
FIG. 6 is a view of Aspergillus niger bacterial pellets at 100-fold and 400-fold microscope respectively;
FIG. 7 is a view of Aspergillus niger bacterial pellets at 100-fold and 400-fold microscope respectively;
FIGS. 1 to 7 show the morphology of Aspergillus niger pellets after citric acid fermentation for 32 hours in each example and comparative example.
Detailed Description
The present invention will be described in further detail with reference to the following examples, but the present invention is not limited thereto in any way. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention, and the following embodiments are all completed by adopting the conventional prior art except for the specific description.
The corn steep liquor used in the following examples had a nitrogen content of 3.7%.
Example 1
1) Culturing Aspergillus niger seeds:
preparing the cultured aspergillus niger spores into aspergillus niger spore suspension by using sterile water for later use;
preparing an Aspergillus niger seed basic culture medium: (1) nutrient salt: KCl 5.5g/L, KH2PO4 12g/L、MgSO4·7H2O 0.35g/L、Na2SO42g/L of anhydrous CaCl25 g/L; (2) trace elements: FeSO4·7H2O 2mg/L、ZnSO4·7H2O 0.5mg/L、MnSO4·H2O 0.2mg/L、CuSO4·5H2O 0.2mg/L、CoCl2·6H2O0.15 mg/L; sterilizing for later use;
simultaneously preparing glucose, corn steep liquor and ammonium sulfate flowing liquid respectively, and sterilizing for later use;
inoculating the aspergillus niger spore suspension into a sterilized seed basal culture medium (the inoculation amount is 35 ten thousand per mL) in an aseptic mode, controlling the seed culture condition to culture the aspergillus niger seeds, wherein the culture condition is as follows: the tank pressure is 0.1Mpa, the tank temperature is 37.2 ℃, the ventilation ratio is 0.20vvm, and the stirring speed is 300 rpm;
controlling the flow acceleration of glucose and corn steep liquor at different culture stages in the culture process: in the Aspergillus niger seed culture stage, the glucose flow acceleration is controlled to be 0.5 g/(L.h), the corn pulp flow acceleration is controlled to be 0.9 g/(L.h), and the ammonium sulfate flow acceleration is controlled to be 0.10 g/(L.h) 0h-8 h; the glucose flow acceleration is controlled to be 1.1 g/(L.h), the corn pulp flow acceleration is controlled to be 1.8 g/(L.h), and the ammonium sulfate flow acceleration is controlled to be 0.16 g/(L.h) within 9h-16 h; the acceleration of the glucose flow is controlled to be 1.8 g/(L.h) and the acceleration of the corn pulp flow is controlled to be 2.8 g/(L.h) within 17h-24 h.
2) And (3) citric acid fermentation process:
preparing a fermentation basal culture medium: (1) carbon source: 184g/L of glucose; (2) nutrient salt: KCl 3.5g/L, KH2PO43g/L、MgSO4·7H2O 0.25g/L、Na2SO40.5g/L anhydrous CaCl27.5 g/L; (3) trace elements: FeSO4·7H2O 5mg/L、ZnSO4·7H2O 2.5mg/L、MnSO4·H2O 0.15mg/L、CuSO4·5H2O 0.1mg/L、CoCl2·6H2O0.1 mg/L, biotin 3.5mg/L, Vb 12 mg/L; sterilizing at high temperature for later use; preparing corn slurry flow liquid, and sterilizing for later use;
after the Aspergillus niger seed liquid is cultured for 24h, the Aspergillus niger seed liquid is aseptically transferred into a fermentation medium (the inoculum size is 3.5 ten thousand/mL) after sterilization and temperature reduction, and fermentation is started under the control of fermentation conditions. The fermentation culture conditions are as follows: the tank pressure is 0.09Mpa, the tank temperature is 37.5 ℃, the ventilation ratio is 0.25vvm, and the stirring speed is 350 rpm;
during the period, the Aspergillus niger form is regulated and controlled by controlling the feeding speed of the nitrogen source (simultaneously limiting the concentrations of phosphorus and manganese) according to different fermentation stages: during the fermentation process of the citric acid, the addition rate of the corn steep liquor is controlled to be 0.41 g/(L.h) for 0h-20h, and the nitrogen source is an organic nitrogen source; 21 h-fermentation is finished, the nitrogen source flow acceleration is controlled to be 0.36/(L.h);
the end-of-fermentation conditions are characterized by a citric acid production rate of less than 0.1g/(100 ml. h).
Under the condition, the cultured Aspergillus niger seeds have the following forms: the fungus balls are compact, regular and small, the hyphae are short and strong, the branching degree is high, and the diameter of the fungus balls is 140-; fermentation parameters: 18.4 percent of total sugar, 54 hours of fermentation period, 15.1g/L of bacteria amount, 18.90 percent of acid production, 102.72 percent of conversion rate, 3.50 g/(L.h) of fermentation index and 117kwh/t of acid consumption for stirring.
Example 2
1) Culturing Aspergillus niger seeds:
preparing the cultured aspergillus niger spores into aspergillus niger spore suspension by using sterile water for later use;
preparing Aspergillus niger seedsBasic culture medium: (1) nutrient salt: KCl 5.5g/L, KH2PO4 13g/L、MgSO4·7H2O 0.35g/L、Na2SO42g/L of anhydrous CaCl25 g/L; (2) trace elements: FeSO4·7H2O 2mg/L、ZnSO4·7H2O 0.5mg/L、MnSO4·H2O 0.25mg/L、CuSO4·5H2O 0.2mg/L、CoCl2·6H2O0.15 mg/L; sterilizing for later use;
simultaneously preparing glucose, corn steep liquor and ammonium sulfate flowing liquid respectively, and sterilizing for later use;
inoculating the aspergillus niger spore suspension into a sterilized seed basal culture medium (the inoculation amount is 35 ten thousand per mL) in an aseptic mode, controlling the seed culture condition to culture the aspergillus niger seeds, wherein the culture condition is as follows: the tank pressure is 0.1Mpa, the tank temperature is 37.2 ℃, the ventilation ratio is 0.20vvm, and the stirring speed is 300 rpm;
controlling the flow acceleration of glucose and corn steep liquor at different culture stages in the culture process: in the Aspergillus niger seed culture stage, the glucose flow acceleration is controlled to be 0.6 g/(L.h), the corn pulp flow acceleration is controlled to be 1.0 g/(L.h), and the ammonium sulfate flow acceleration is controlled to be 0.11 g/(L.h) 0h-8 h; the glucose flow acceleration is controlled to be 1.2 g/(L.h), the corn pulp flow acceleration is controlled to be 1.9 g/(L.h), and the ammonium sulfate flow acceleration is controlled to be 0.18 g/(L.h) within 9h-16 h; the acceleration of the glucose flow is controlled to be 1.9 g/(L.h) and the acceleration of the corn pulp flow is controlled to be 3.0 g/(L.h) within 17h-24 h;
2) and (3) citric acid fermentation process:
preparing a fermentation basal culture medium: (1) carbon source: 184g/L of glucose; (2) nutrient salt: KCl 3.5g/L, KH2PO4 2.5g/L、MgSO4·7H2O 0.25g/L、Na2SO40.5g/L anhydrous CaCl27.5 g/L; (3) trace elements: FeSO4·7H2O 5mg/L、ZnSO4·7H2O 2.5mg/L、MnSO4·H2O 0.17mg/L、CuSO4·5H2O 0.1mg/L、CoCl2·6H2O0.1 mg/L, biotin 3.0mg/L, Vb12.2mg/L; sterilizing at high temperature for later use; preparing corn slurry flow liquid, and sterilizing for later use;
after the Aspergillus niger seed liquid is cultured for 24h, the Aspergillus niger seed liquid is aseptically transferred into a fermentation medium (the inoculum size is 3.5 ten thousand/mL) after sterilization and temperature reduction, and fermentation is started under the control of fermentation conditions. The fermentation culture conditions are as follows: the tank pressure is 0.09Mpa, the tank temperature is 37.5 ℃, the ventilation ratio is 0.25vvm, and the stirring speed is 350 rpm;
during the period, the Aspergillus niger form is regulated and controlled by controlling the feeding speed of the nitrogen source (simultaneously limiting the concentrations of phosphorus and manganese) according to different fermentation stages: during the fermentation process of the citric acid, the addition rate of the corn steep liquor is controlled to be 0.42 g/(L.h) 0h-20h, and the nitrogen source is an organic nitrogen source; 21 h-fermentation is finished, the nitrogen source flow acceleration is controlled to be 0.37/(L.h);
the end-of-fermentation conditions are characterized by a citric acid production rate of less than 0.1g/(100 ml. h).
Under the condition, the cultured Aspergillus niger seeds have the following forms: the fungus balls are compact, regular and small, the hyphae are short and strong, the branching degree is high, and the diameter of the fungus balls is 145-; fermentation parameters: 18.4 percent, a fermentation period of 54 hours, a bacterial amount of 15.5g/L, an acid production of 18.95 percent, a conversion rate of 102.99 percent, a fermentation index of 3.51 g/(L.h), and stirring power consumption of 118kwh/t acid.
Example 3
1) Culturing Aspergillus niger seeds:
preparing the cultured aspergillus niger spores into aspergillus niger spore suspension by using sterile water for later use;
preparing an Aspergillus niger seed basic culture medium: (1) nutrient salt: KCl 5.5g/L, KH2PO4 14g/L、MgSO4·7H2O 0.35g/L、Na2SO42g/L of anhydrous CaCl25 g/L; (2) trace elements: FeSO4·7H2O 2mg/L、ZnSO4·7H2O 0.5mg/L、MnSO4·H2O 0.30mg/L、CuSO4·5H2O 0.2mg/L、CoCl2·6H2O0.15 mg/L; sterilizing for later use;
simultaneously preparing glucose, corn steep liquor and ammonium sulfate flowing liquid respectively, and sterilizing for later use;
inoculating the aspergillus niger spore suspension into a sterilized seed basal culture medium (the inoculation amount is 35 ten thousand per mL) in an aseptic mode, controlling the seed culture condition to culture the aspergillus niger seeds, wherein the culture condition is as follows: the tank pressure is 0.1Mpa, the tank temperature is 37.2 ℃, the ventilation ratio is 0.20vvm, and the stirring speed is 300 rpm;
controlling the flow acceleration of glucose and corn steep liquor at different culture stages in the culture process: in the Aspergillus niger seed culture stage, the glucose flow acceleration is controlled to be 0.7 g/(L.h), the corn pulp flow acceleration is controlled to be 1.1 g/(L.h), and the ammonium sulfate flow acceleration is controlled to be 0.12 g/(L.h) 0h-8 h; the glucose flow acceleration is controlled to be 1.3 g/(L.h), the corn pulp flow acceleration is controlled to be 2.0 g/(L.h), and the ammonium sulfate flow acceleration is controlled to be 0.20 g/(L.h) within 9h-16 h; the acceleration of the glucose flow is controlled to be 2.0 g/(L.h) and the acceleration of the corn pulp flow is controlled to be 3.1 g/(L.h) within 17h-24 h.
2) And (3) citric acid fermentation process:
preparing a fermentation basal culture medium: (1) carbon source: 185g/L of glucose; (2) nutrient salt: KCl 3.5g/L, KH2PO42.7g/L、MgSO4·7H2O 0.25g/L、Na2SO40.5g/L anhydrous CaCl27.5 g/L; (3) trace elements: FeSO4·7H2O 5mg/L、ZnSO4·7H2O 2.5mg/L、MnSO4·H2O 0.15mg/L、CuSO4·5H2O 0.1mg/L、CoCl2·6H2O0.1 mg/L, biotin 2.8mg/L, Vb12.3mg/L; sterilizing at high temperature for later use; preparing corn slurry flow liquid, and sterilizing for later use;
after the Aspergillus niger seed liquid is cultured for 24h, the Aspergillus niger seed liquid is aseptically transferred into a fermentation medium (the inoculum size is 3.5 ten thousand/mL) after sterilization and temperature reduction, and fermentation is started under the control of fermentation conditions. The fermentation culture conditions are as follows: the tank pressure is 0.09Mpa, the tank temperature is 37.5 ℃, the ventilation ratio is 0.25vvm, and the stirring speed is 350 rpm;
during the period, the Aspergillus niger form is regulated and controlled by controlling the feeding speed of the nitrogen source (simultaneously limiting the concentrations of phosphorus and manganese) according to different fermentation stages: during the fermentation process of the citric acid, the addition rate of the corn steep liquor is controlled to be 0.43 g/(L.h) within 0h-20h, and the nitrogen source is an organic nitrogen source; 21 h-fermentation is finished, the nitrogen source flow acceleration is controlled to be 0.38/(L.h);
the end-of-fermentation conditions are characterized by a citric acid production rate of less than 0.1g/(100 ml. h).
Under the condition, the cultured Aspergillus niger seeds have the following forms: the fungus balls are compact, regular and small, the hyphae are short and strong, the branching degree is high, and the diameter of the fungus balls is 145-; fermentation parameters: 18.5 percent, a fermentation period of 54 hours, a bacterial amount of 15.8g/L, an acid production of 19.00 percent, a conversion rate of 102.70 percent, a fermentation index of 3.52 g/(L.h), and stirring power consumption of 118kwh/t acid.
Example 4
1) Culturing Aspergillus niger seeds:
preparing the cultured aspergillus niger spores into aspergillus niger spore suspension by using sterile water for later use;
preparing an Aspergillus niger seed basic culture medium: (1) nutrient salt: KCl 5.5g/L, KH2PO4 12.5g/L、MgSO4·7H2O 0.35g/L、Na2SO42g/L of anhydrous CaCl25 g/L; (2) trace elements: FeSO4·7H2O 2mg/L、ZnSO4·7H2O 0.5mg/L、MnSO4·H2O 0.25mg/L、CuSO4·5H2O 0.2mg/L、CoCl2·6H2O0.15 mg/L; sterilizing for later use;
simultaneously preparing glucose, corn steep liquor and ammonium sulfate flowing liquid respectively, and sterilizing for later use;
inoculating the aspergillus niger spore suspension into a sterilized seed basal culture medium (the inoculation amount is 35 ten thousand per mL) in an aseptic mode, controlling the seed culture condition to culture the aspergillus niger seeds, wherein the culture condition is as follows: the tank pressure is 0.1Mpa, the tank temperature is 37.2 ℃, the ventilation ratio is 0.20vvm, and the stirring speed is 300 rpm;
controlling the flow acceleration of glucose and corn steep liquor at different culture stages in the culture process: in the Aspergillus niger seed culture stage, the glucose flow acceleration is controlled to be 0.55 g/(L.h), the corn pulp flow acceleration is controlled to be 0.95 g/(L.h), and the ammonium sulfate flow acceleration is controlled to be 0.11 g/(L.h) 0h-8 h; the glucose flow acceleration is controlled to be 1.15 g/(L.h), the corn pulp flow acceleration is controlled to be 1.9 g/(L.h), and the ammonium sulfate flow acceleration is controlled to be 0.17 g/(L.h) within 9h-16 h; the acceleration of the glucose flow is controlled to be 1.9 g/(L.h) and the acceleration of the corn pulp flow is controlled to be 2.9 g/(L.h) within 17h-24 h.
2) And (3) citric acid fermentation process:
preparing a fermentation basal culture medium: (1) carbon source: 185g/L of glucose; (2) nutrient salt: KCl 3.5g/L, KH2PO43.1g/L、MgSO4·7H2O 0.25g/L、Na2SO40.5g/L anhydrous CaCl27.5 g/L; (3) trace elements: FeSO4·7H2O 5mg/L、ZnSO4·7H2O 2.5mg/L、MnSO4·H2O 0.16mg/L、CuSO4·5H2O 0.1mg/L、CoCl2·6H2O0.1 mg/L, biotin 3.2mg/L, Vb 11.9mg/L; sterilizing at high temperature for later use; preparing corn slurry flow liquid, and sterilizing for later use;
after the Aspergillus niger seed liquid is cultured for 24h, the Aspergillus niger seed liquid is aseptically transferred into a fermentation medium (the inoculum size is 3.5 ten thousand/mL) after sterilization and temperature reduction, and fermentation is started under the control of fermentation conditions. The fermentation culture conditions are as follows: the tank pressure is 0.09Mpa, the tank temperature is 37.5 ℃, the ventilation ratio is 0.25vvm, and the stirring speed is 350 rpm;
during the period, the Aspergillus niger form is regulated and controlled by controlling the feeding speed of the nitrogen source (simultaneously limiting the concentrations of phosphorus and manganese) according to different fermentation stages: during the fermentation process of citric acid, the addition rate of the corn steep liquor is controlled to be 0.44 g/(L.h) 0h-20h, and the nitrogen source is an organic nitrogen source; 21 h-fermentation is finished, the nitrogen source flow acceleration is controlled to be 0.39/(L.h);
the end-of-fermentation conditions are characterized by a citric acid production rate of less than 0.1g/(100 ml. h).
Under the condition, the cultured Aspergillus niger seeds have the following forms: the fungus balls are compact, regular and small, the hyphae are short and strong, the branching degree is high, and the diameter of the fungus balls is 140-; fermentation parameters: 18.5 percent, a fermentation period of 55 hours, a bacterial load of 15.3g/L, an acid yield of 19.10 percent, a conversion rate of 103.24 percent, a fermentation index of 3.47 g/(L.h), and stirring power consumption of 121kwh/t acid.
Example 5
1) Culturing Aspergillus niger seeds:
preparing the cultured aspergillus niger spores into aspergillus niger spore suspension by using sterile water for later use;
preparing an Aspergillus niger seed basic culture medium: (1) nutrient salt: KCl 5.5g/L, KH2PO4 13.0g/L、MgSO4·7H2O 0.35g/L、Na2SO42g/L of anhydrous CaCl25 g/L; (2) trace elements: FeSO4·7H2O 2mg/L、ZnSO4·7H2O 0.5mg/L、MnSO4·H2O 0.27mg/L、CuSO4·5H2O 0.2mg/L、CoCl2·6H2O0.15 mg/L; sterilizing for later use;
simultaneously preparing glucose, corn steep liquor and ammonium sulfate flowing liquid respectively, and sterilizing for later use;
inoculating the aspergillus niger spore suspension into a sterilized seed basal culture medium (the inoculation amount is 35 ten thousand per mL) in an aseptic mode, controlling the seed culture condition to culture the aspergillus niger seeds, wherein the culture condition is as follows: the tank pressure is 0.1Mpa, the tank temperature is 37.2 ℃, the ventilation ratio is 0.20vvm, and the stirring speed is 300 rpm;
controlling the flow acceleration of glucose and corn steep liquor at different culture stages in the culture process: in the Aspergillus niger seed culture stage, the glucose flow acceleration is controlled to be 0.65 g/(L.h), the corn pulp flow acceleration is controlled to be 1.05 g/(L.h), and the ammonium sulfate flow acceleration is controlled to be 0.11 g/(L.h) 0h-8 h; the glucose flow acceleration is controlled to be 1.25 g/(L.h), the corn pulp flow acceleration is controlled to be 1.95 g/(L.h), and the ammonium sulfate flow acceleration is controlled to be 0.18 g/(L.h) within 9h-16 h; the acceleration of the glucose flow is controlled to be 1.90 g/(L.h) and the acceleration of the corn pulp flow is controlled to be 3.0 g/(L.h) within 17h-24 h;
2) and (3) citric acid fermentation process:
preparing a fermentation basal culture medium: (1) carbon source: 185g/L of glucose; (2) nutrient salt: KCl 3.5g/L, KH2PO43.2g/L、MgSO4·7H2O 0.25g/L、Na2SO40.5g/L anhydrous CaCl27.5 g/L; (3) trace elements: FeSO4·7H2O 5mg/L、ZnSO4·7H2O 2.5mg/L、MnSO4·H2O 0.14mg/L、CuSO4·5H2O 0.1mg/L、CoCl2·6H2O0.1 mg/L, biotin 3.7mg/L, Vb 12.1mg/L; sterilizing at high temperature for later use; preparing corn slurry flow liquid, and sterilizing for later use;
after the Aspergillus niger seed liquid is cultured for 24h, the Aspergillus niger seed liquid is aseptically transferred into a fermentation medium (the inoculum size is 3.5 ten thousand/mL) after sterilization and temperature reduction, and fermentation is started under the control of fermentation conditions. The fermentation culture conditions are as follows: the tank pressure is 0.09Mpa, the tank temperature is 37.5 ℃, the ventilation ratio is 0.25vvm, and the stirring speed is 350 rpm;
during the period, the Aspergillus niger form is regulated and controlled by controlling the feeding speed of the nitrogen source (simultaneously limiting the concentrations of phosphorus and manganese) according to different fermentation stages: during the fermentation process of the citric acid, the addition rate of the corn steep liquor is controlled to be 0.45 g/(L.h) for 0h-20h, and the nitrogen source is an organic nitrogen source; 21 h-fermentation is finished, the nitrogen source flow acceleration is controlled to be 0.40 g/(L.h);
the end-of-fermentation conditions are characterized by a citric acid production rate of less than 0.1g/(100 ml. h).
Under the condition, the cultured Aspergillus niger seeds have the following forms: the fungus balls are compact, regular and small, the hyphae are short and strong, the branching degree is high, and the diameter of the fungus balls is 140-; fermentation parameters: 18.5 percent, a fermentation period of 54 hours, a bacterial load of 15.2g/L, an acid production of 19.05 percent, a conversion rate of 102.97 percent, a fermentation index of 3.53 g/(L.h), and stirring power consumption of 118kwh/t acid.
Comparative example 1
1) Culturing Aspergillus niger seeds:
preparing the cultured aspergillus niger spores into aspergillus niger spore suspension by using sterile water for later use;
preparing an Aspergillus niger seed culture medium: (1) carbon source: glucose 27.2g/L, nitrogen source: 44g/L of corn steep liquor and 2.08g/L of ammonium sulfate; (2) nutrient salt: KCl 5.5g/L, KH2PO4 11g/L、MgSO4·7H2O 0.35g/L、Na2SO42g/L of anhydrous CaCl25 g/L; (3) trace elements: FeSO4·7H2O 2mg/L、ZnSO4·7H2O 0.5mg/L、MnSO4·H2O 0.15mg/L、CuSO4·5H2O 0.2mg/L、CoCl2·6H2O0.15 mg/L; sterilizing for later use;
inoculating the Aspergillus niger spore suspension into a sterilized seed culture medium (the inoculum size is 35 ten thousand/mL) in an aseptic mode, and controlling the seed culture conditions to carry out Aspergillus niger seed culture, wherein the culture conditions are as follows: the pot pressure was 0.1MPa, the pot temperature was 37.2 ℃, the aeration ratio was 0.20vvm, and the stirring speed was 300 rpm.
2) And (3) citric acid fermentation process:
preparing a fermentation medium: (1) carbon source: glucose 185g/L, nitrogen source: 23g/L of corn steep liquor; (2) nutrient salt: KCl 3.5g/L, KH2PO4 2g/L、MgSO4·7H2O 0.25g/L、Na2SO40.5g/L anhydrous CaCl27.5 g/L; (3) trace elements: FeSO4·7H2O 5mg/L、ZnSO4·7H2O 2.5mg/L、MnSO4·H2O 0.11mg/L、CuSO4·5H2O 0.1mg/L、CoCl2·6H2O0.1 mg/L, biotin 3.5mg/L, Vb 12 mg/L; and (5) sterilizing at high temperature for later use.
After the Aspergillus niger seed liquid is cultured for 24h, the Aspergillus niger seed liquid is aseptically transferred into a fermentation medium (the inoculum size is 3.5 ten thousand/mL) after sterilization and temperature reduction, and fermentation is started under the control of fermentation conditions. The fermentation culture conditions are as follows: the tank pressure is 0.09Mpa, the tank temperature is 37.5 ℃, the ventilation ratio is 0.25vvm, and the stirring speed is 350 rpm; the end-of-fermentation conditions are characterized by a citric acid production rate of less than 0.1g/(100 ml. h).
Under the condition, the cultured Aspergillus niger seeds have the following forms: the mycelium pellet is loose, irregular and large, the mycelium is slender and has less branching degree, and the diameter of the mycelium pellet is 180-; fermentation parameters: the total sugar content is 18.5%, the fermentation period is 63h, the bacterial load is 18.6g/L, the acid production is 18.70%, the conversion rate is 101.08%, the fermentation index is 2.97 g/(L.h), and the stirring power consumption is 154kwh/t acid.
Comparative example 2
1) Culturing Aspergillus niger seeds:
preparing the cultured aspergillus niger spores into aspergillus niger spore suspension by using sterile water for later use;
preparing an Aspergillus niger seed culture medium: (1) carbon source: glucose 29.2g/L, nitrogen source: 47.2g/L of corn steep liquor and 2.32g/L of ammonium sulfate; (2) nutrient salt: KCl 5.5g/L, KH2PO4 15g/L、MgSO4·7H2O 0.35g/L、Na2SO42g/L of anhydrous CaCl25 g/L; (3) trace elements: FeSO4·7H2O 2mg/L、ZnSO4·7H2O 0.5mg/L、MnSO4·H2O 0.35mg/L、CuSO4·5H2O 0.2mg/L、CoCl2·6H2O0.15 mg/L; sterilizing for later use;
inoculating the Aspergillus niger spore suspension into a sterilized seed culture medium (the inoculum size is 35 ten thousand/mL) in an aseptic mode, and controlling the seed culture conditions to carry out Aspergillus niger seed culture, wherein the culture conditions are as follows: the pot pressure was 0.1MPa, the pot temperature was 37.2 ℃, the aeration ratio was 0.20vvm, and the stirring speed was 300 rpm.
2) And (3) citric acid fermentation process:
preparing a fermentation medium: (1) carbon source: glucose 175g/L, nitrogen source: 24g/L of corn steep liquor; (2) nutrient salt: KCl 3.5g/L, KH2PO4 4g/L、MgSO4·7H2O 0.25g/L、Na2SO40.5g/L anhydrous CaCl27.5 g/L; (3) trace elements: FeSO4·7H2O 5mg/L、ZnSO4·7H2O 2.5mg/L、MnSO4·H2O 0.19mg/L、CuSO4·5H2O 0.1mg/L、CoCl2·6H2O0.1 mg/L, biotin 3.5mg/L, Vb 12 mg/L; and (5) sterilizing at high temperature for later use.
After the Aspergillus niger seed liquid is cultured for 24h, the Aspergillus niger seed liquid is aseptically transferred into a fermentation medium (the inoculum size is 3.5 ten thousand/mL) after sterilization and temperature reduction, and fermentation is started under the control of fermentation conditions. The fermentation culture conditions are as follows: the tank pressure is 0.09Mpa, the tank temperature is 37.5 ℃, the ventilation ratio is 0.25vvm, and the stirring speed is 350 rpm; the end-of-fermentation conditions are characterized by a citric acid production rate of less than 0.1g/(100 ml. h).
Under the condition, the cultured Aspergillus niger seeds have the following forms: the bacterial balls are loose, irregular and large, the hyphae are slender and the branching degree is less, and the diameter of the bacterial balls is 190-; fermentation parameters: 17.5 percent of total sugar, 58 hours of fermentation period, 19.1g/L of bacteria amount, 17.80 percent of acid production, 101.71 percent of conversion rate, 3.07 g/(L.h) of fermentation index and 135kwh/t of acid consumption for stirring.
Comparing the aspergillus niger morphology (the size of the cell pellet morphology is stable already when the citric acid is cultured for 32 h) in the step 2) of citric acid fermentation for 32h and various fermentation parameters in the fermentation process in each embodiment and comparative example, and detailed descriptions are shown in tables 1 and 2:
table 1: the invention is compared with the Aspergillus niger form in the prior art
Table 2: the invention compares the citric acid fermentation parameters with the prior art
Through the comparison, compared with the prior art, the novel process for culturing and fermenting citric acid by aspergillus niger disclosed by the invention comprises the following steps: (1) the fermentation period is shortened, the sugar-acid conversion rate is improved, and the fermentation index is improved; (2) the Aspergillus niger is small in fungus ball shape, short in hypha and low in stirring power consumption; (3) the microbial biomass is low, and the yield of citric acid is correspondingly improved. Therefore, the industrial competitiveness of the citric acid is comprehensively improved.
Claims (8)
1. A method for regulating and controlling citric acid fermentation by controlling the morphology of Aspergillus niger mycelia comprises 1) Aspergillus niger seed culture and 2) citric acid fermentation process, and is characterized in that in the step 1) Aspergillus niger seed culture process:
the phosphorus element concentration in the seed basic culture medium is as follows: 2.5-3.5g/L, the concentration of manganese element is as follows: 0.06-0.10 mg/L;
the Aspergillus niger seed culture stage controls the flow rate of a carbon source and a nitrogen source to regulate the Aspergillus niger form, and specifically comprises the following steps: controlling the carbon source flow acceleration rate to be 0.5-0.7g sugar/(L.h) and the nitrogen source flow acceleration rate to be 55-66 mg/(L.h), wherein the ratio of inorganic nitrogen to organic nitrogen is 0.5:1-0.8:1, and the C/N ratio is 3.0:1-5.2: 1; the carbon source flow rate is controlled to be 1.1-1.3g sugar/(L.h) in 9h-16h, and the nitrogen source flow rate is controlled to be 100-120 mg/(L.h), wherein the ratio of inorganic nitrogen to organic nitrogen is 0.4:1-0.65:1, and the C/N is 3.6:1-5.2: 1; the carbon source flow rate is controlled to be 1.8-2.0g sugar/(L.h) within 17h-24h, the nitrogen source flow rate is controlled to be 100-120 mg/(L.h), wherein the nitrogen sources are all organic nitrogen sources, and the C/N is 6.0:1-7.8: 1;
the specific process comprises the following steps: inoculating Aspergillus niger spores into a seed basal culture medium, and controlling seed culture conditions to culture Aspergillus niger seeds, wherein the seed culture conditions are as follows: the pressure in the tank is 0.06-0.12Mpa, the temperature in the tank is 32-40 ℃, the ventilation ratio is 0.15-0.23vvm, and the stirring speed is 250-350 rpm.
2. The method for regulating and controlling citric acid fermentation by controlling aspergillus niger bacteria pellet morphology as claimed in claim 1, wherein, in the step 2) citric acid fermentation process:
in the fermentation basic culture medium, the concentration of phosphorus element is as follows: 0.55-0.80g/L, manganese element concentration is as follows: 0.04-0.06 mg/L;
controlling the feeding speed of the nitrogen source according to different fermentation stages to regulate the shape of the Aspergillus niger: controlling the nitrogen source flow acceleration rate to be 15-17 mg/(L.h) for 0h-20h, wherein the nitrogen source is an organic nitrogen source; 21 h-fermentation is finished, the adding speed of the nitrogen source flow is controlled to be 13-15 mg/(L.h), and the nitrogen source is an organic nitrogen source;
the specific process comprises the following steps: after the Aspergillus niger seed liquid is cultured, transferring the seed liquid into a fermentation basal culture medium, and controlling the fermentation condition to start fermentation; the fermentation culture conditions are as follows: the pressure in the tank is 0.06-0.12Mpa, the temperature in the tank is 35-40 ℃, the ventilation ratio is 0.18-0.25vvm, and the stirring speed is 300-400 rpm.
3. The method for regulating and controlling citric acid fermentation by controlling aspergillus niger bacteria pellet morphology as claimed in claim 1, wherein in step 1) aspergillus niger seed culture stage, aspergillus niger spore inoculation concentration is 25-50 ten thousand/mL.
4. The method for regulating and controlling citric acid fermentation by controlling aspergillus niger bacteria ball shape according to claim 1, wherein, in the aspergillus niger seed culture process of step 1), the seed basal medium consists of:
(1) nutrient salt: KCl 4-6g/L, phosphorus-containing nutrient salt (phosphorus concentration: 2.5-3.5g/L), MgSO4·7H2O0.25-0.4g/L、Na2SO42g/L of anhydrous CaCl2 4-6g/L；
(2) Trace elements: FeSO4·7H2O 1.5-2.5mg/L、ZnSO4·7H20.4-0.6mg/L of O, manganese-containing nutrient salt (the concentration of manganese element is controlled to be 0.06-0.10mg/L), and CuSO4·5H2O 0.15-0.25mg/L、CoCl2·6H2O 0.10-0.20mg/L。
5. The method for regulating and controlling citric acid fermentation by controlling aspergillus niger bacteria pellet morphology according to claim 1 or 2, characterized in that the inoculation concentration of the bacteria pellet during the citric acid fermentation process of step 2) is controlled to be 2.5-5.0 ten thousand/mL.
6. The method for regulating citric acid fermentation by controlling aspergillus niger bacteria pellet morphology as claimed in claim 2, wherein said fermentation basal medium consists of:
(1) carbon source: total sugar: 175-185 g/L;
(2) nutrient salt: KCl 2.5-4.0g/L, phosphorus-containing nutrient salt (the concentration of phosphorus element is controlled to be 0.55-0.80g/L), MgSO4·7H2O 0.15-0.35g/L、Na2SO40.40-0.55g/L of anhydrous CaCl2 6.0-8.0g/L；
(3) Trace elements: FeSO4·7H2O 4.0-6.0mg/L、ZnSO4·7H2O2.0-3.0 mg/L, manganese-containing nutrient salt (the concentration of manganese element is controlled to be 0.04-0.06mg/L), CuSO4·5H2O 0.08-0.12mg/L、CoCl2·6H2O0.07-0.11 mg/L, biotin 2.5-4.0mg/L, and Vb 11.8-2.5 mg/L.
7. The method for regulating and controlling citric acid fermentation by controlling aspergillus niger bacteria ball shape according to claim 1 or 2, wherein the carbon source refers to one or more of starch and glucose; the inorganic nitrogen refers to one or more of ammonia water, ammonium salt, nitrate and urea; the organic nitrogen is one or more of corn steep liquor, bean cake powder, peanut powder and yeast extract.
8. The method for regulating citric acid fermentation by controlling aspergillus niger bacteria pellet morphology according to claim 4 or 6, wherein the phosphorus-containing nutrient salt refers to one or more of potassium dihydrogen phosphate and dipotassium hydrogen phosphate; the manganese-containing nutrient salt refers to one or more of manganese sulfate and manganese chloride.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103290070A (en) * | 2013-06-20 | 2013-09-11 | 南通凯赛生化工程设备有限公司 | Method for producing citric acid through continuous batch feeding fermentation |
| CN105385711A (en) * | 2015-12-28 | 2016-03-09 | 日照金禾博源生化有限公司 | Citric acid fermentation technology |
| CN108315363A (en) * | 2018-05-15 | 2018-07-24 | 日照金禾博源生化有限公司 | A kind of citric acid fermentation method for persistently mending nitrogen |
| CN109207373A (en) * | 2018-09-21 | 2019-01-15 | 天津科技大学 | The method that one plant height produces the microbial strains and its fermentation starch saccharic production citric acid of citric acid |
| WO2020042481A1 (en) * | 2018-08-28 | 2020-03-05 | 江苏国信协联能源有限公司 | Method for continuously culturing aspergillus niger seeds and producing citric acid using same |
-
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- 2021-09-30 CN CN202111168793.4A patent/CN114045313B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103290070A (en) * | 2013-06-20 | 2013-09-11 | 南通凯赛生化工程设备有限公司 | Method for producing citric acid through continuous batch feeding fermentation |
| CN105385711A (en) * | 2015-12-28 | 2016-03-09 | 日照金禾博源生化有限公司 | Citric acid fermentation technology |
| CN108315363A (en) * | 2018-05-15 | 2018-07-24 | 日照金禾博源生化有限公司 | A kind of citric acid fermentation method for persistently mending nitrogen |
| WO2020042481A1 (en) * | 2018-08-28 | 2020-03-05 | 江苏国信协联能源有限公司 | Method for continuously culturing aspergillus niger seeds and producing citric acid using same |
| CN109207373A (en) * | 2018-09-21 | 2019-01-15 | 天津科技大学 | The method that one plant height produces the microbial strains and its fermentation starch saccharic production citric acid of citric acid |
Non-Patent Citations (3)
| Title |
|---|
| M. PAPAGIANNI 等: "The influence of glucose concentration on citric acid production and morphology of Aspergillus niger in batch and culture", ENZYME AND MICROBIAL TECHNOLOGY, vol. 25, no. 8, pages 710 - 717 * |
| 刘会等: "菌球形态与柠檬酸产量相关性研究", 现代农业科技, vol. 2020, no. 20, pages 201 - 202 * |
| 杨儒文等: "黑曲霉菌球形态对产酸影响", 当代化工, vol. 49, no. 8, pages 1689 - 1693 * |
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