CN106906122B - System and method for co-producing propionic acid and salt thereof and succinic acid and salt thereof - Google Patents

System and method for co-producing propionic acid and salt thereof and succinic acid and salt thereof Download PDF

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CN106906122B
CN106906122B CN201710320439.6A CN201710320439A CN106906122B CN 106906122 B CN106906122 B CN 106906122B CN 201710320439 A CN201710320439 A CN 201710320439A CN 106906122 B CN106906122 B CN 106906122B
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propionic acid
acid
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CN106906122A (en
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王自强
徐国霞
李小连
王云山
苏志国
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M43/00—Combinations of bioreactors or fermenters with other apparatus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/44—Polycarboxylic acids
    • C12P7/46—Dicarboxylic acids having four or less carbon atoms, e.g. fumaric acid, maleic acid
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/52—Propionic acid; Butyric acids

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Abstract

The invention relates to a system and a method for co-producing propionic acid and salts thereof and succinic acid and salts thereof, wherein the method comprises the following steps: (1) Culturing propionibacterium acidogenesis in a fermentation medium, and then transferring the propionibacterium acidogenesis in a fermentation tank for fermentation; (2) Introducing the fermentation liquor into a membrane separation unit, filtering out fermentation clear liquid, and returning the somatic cells to the step (1); (3) Acidifying the fermentation clear liquid obtained in the step (2) in an acidification unit until the pH value is less than or equal to 3.0; (4) Loading the acidified fermentation liquor to a first chromatographic unit to obtain a first permeate; (5) Loading the first permeate to a second chromatographic unit, and returning the obtained second permeate to the step (1); (6) eluting to obtain succinic acid and salts thereof, propionic acid and salts thereof. The invention realizes the co-production of propionic acid and succinic acid, and the production efficiency of the propionic acid and the succinic acid is respectively more than 0.440 g/(L.h) and more than 0.147 g/(L.h).

Description

System and method for co-producing propionic acid and salt thereof and succinic acid and salt thereof
Technical Field
The invention relates to the field of bioengineering, in particular to a system and a method for co-producing propionic acid and salts thereof and succinic acid and salts thereof.
Background
Propionibacterium acidogenesis (Propionibacterium acidipropionici) is one of the main strains for producing propionic acid by anaerobic fermentation, but in the fermentation process, continuous accumulation of propionic acid can produce feedback inhibition effect on cell growth and propionic acid metabolism, so that fermentation efficiency is reduced. For example, when the propionic acid concentration in the fermentation liquid is more than 10g/L, the growth rate of the cells starts to slow down, and when the propionic acid concentration reaches 35g/L, the growth rate of the cells drops sharply. To relieve the feedback inhibition of propionic acid on fermentation, researchers genetically modified the production strain (Biotechnology and Bioengineering,2009,104:766-773;Metabolic Engineering,2015,27:46-56), studied the feedback inhibition mechanism of propionic acid (Journal of Biotechnology,2013, 167:56-63), and adopted appropriate regulatory strategies (Bioresource Technology,2012,105:128-133;Bioresource Technology,2013,135:504-512) to increase the yield of propionic acid in the batch fermentation process. In addition, researches also find that the propionic acid is removed in time by adopting a process engineering means in the fermentation process, so that the feedback regulation effect of the propionic acid on the fermentation can be effectively slowed down or eliminated, and the fermentation efficiency is improved. Goswami constructed an in situ cell-retaining reactor, which retained cells by stainless steel rotary filters and filtered fermentation supernatants containing high concentrations of propionic acid, thereby achieving separation of cells from propionic acid (Applied Microbiology and Biotechnology,2001, 56:676-680). The university of Nanjing industry, ouyangping Kaiser, constructs a porous fiber bed reactor to timely separate propionic acid generated in the fermentation process, so that the fermentation yield is improved (Journal of Biotechnology,2012, 164:202-210). University of Zhejiang Xu Zhina teaches that fermentation efficiency is improved by immobilizing somatic cells on a fiber bed reactor with fed-batch glucose (Bioresource Technology,2012, 112:248-253). The institute of Chinese sciences Wang Yunshan researchers coupled expanded bed chromatography with fermentation, established a novel fermentation process featuring expanded bed in situ adsorption, and improved propionic acid yield (Bioresource Technology,2012, 104:652-659). However, in the industrial production process, the expanded bed has low filling efficiency, and the process generally has the problems of difficult scale-up, high cost and the like.
In addition, the fermentation broth contains a large amount of succinic acid, lactic acid, acetic acid and the like in addition to propionic acid, so that the purity of propionic acid is limited. CN103667373a discloses a method for producing propionic acid and salts thereof and co-producing succinic acid and salts thereof by microbial fermentation, which uses carbonate to neutralize propionic acid, reduce the inhibition of propionic acid to cell growth, and the produced carbon dioxide is used for synthesizing succinic acid, so that the succinic acid output is mainly improved, but the process does not effectively separate succinic acid and propionic acid in fermentation liquor, and the product is mainly a mixture of calcium succinate and calcium propionate.
Therefore, there is an urgent need to develop a process for co-production of propionic acid and its salts and succinic acid and its salts, which can effectively improve the purity of propionic acid while being easy to industrialize.
Disclosure of Invention
In view of the problems existing in the prior art, in order to realize the co-production of propionic acid and salts thereof and succinic acid and salts thereof, the invention adopts the following technical scheme that the fermentation efficiency and the product purity are improved, and the industrialization difficulty and cost are reduced:
in a first aspect, the invention provides a system for co-producing propionic acid and salts thereof, and succinic acid and salts thereof, the system comprising a fermentation unit, a membrane separation unit, an acidification unit, a first chromatography unit and a second chromatography unit which are connected in sequence;
A first loop is arranged between the membrane separation unit and the fermentation unit;
a second loop is arranged between the second chromatography unit and the fermentation unit.
The method comprises the steps of fermenting somatic cells in a fermentation unit to generate propionic acid and succinic acid, enabling fermentation liquor containing the somatic cells, nutrient substances, the propionic acid and the succinic acid to pass through a membrane separation unit, enabling the separated somatic cells to flow back to the fermentation unit through a first loop, enabling separated fermentation clear liquor to enter an acidification unit for acidification to obtain acidified fermentation clear liquor, enabling the acidified fermentation clear liquor to enter a first chromatographic unit, enabling the first chromatographic unit to adsorb the succinic acid with higher acidity, enabling permeate liquor to enter a second chromatographic unit, enabling the second chromatographic unit to adsorb the propionic acid, enabling the permeate liquor to contain the nutrient substances, and enabling the permeate liquor to flow back to the fermentation unit through the second loop to continue fermentation.
The fermentation unit according to the invention preferably comprises a fermenter.
Preferably, the material of the filtration membrane in the membrane separation unit comprises polyethersulfone.
Preferably, the pore size of the filtration membrane in the membrane separation unit is 0.1 to 0.22 μm, for example 0.1 μm or 0.22 μm, etc., preferably 0.22 μm.
Preferably, the membrane separation unit is passed throughThe flux per unit area of the filter membrane is 20-30L/(h.m) 2 ) For example 20L/(h.m) 2 )、21L/(h·m 2 )、22L/(h·m 2 )、23L/(h·m 2 )、24L/(h·m 2 )、25L/(h·m 2 )、26L/(h·m 2 )、27L/(h·m 2 )、28L/(h·m 2 )、29L/(h·m 2 ) Or 30L/(h.m) 2 ) Etc.
Preferably, the filtering area of the filtering membrane in the membrane separation unit is more than or equal to 0.12m 2 For example 0.12m 2 、0.2m 2 、0.5m 2 、1m 2 、2.5m 2 、5m 2 Or 10m 2 Etc., preferably 0.12m 2 ～0.50m 2 。
Preferably, the acidification unit is filled with a cation exchange resin.
Preferably, the acidification unit is filled with an H-type cation exchange resin.
Preferably, the volume filling factor of the cation exchange resin in the acidification unit is between 0.7 and 0.8, for example 0.7, 0.71, 0.73, 0.75, 0.78 or 0.8, etc.
Preferably, the volume ratio of the acidification unit to the fermentation unit is 1 (2-4), e.g. 1:2, 1:2.2, 1:2.5, 1:2.8, 1:3, 1:3.2, 1:3.5, 1:3.8 or 1:4, etc., preferably 1 (3-4).
Preferably, the first chromatography unit is filled with an anion exchange resin, preferably ZGD630 anion exchange resin.
Preferably, the volume filling factor of the anion exchange resin in the first chromatography unit is 0.7 to 0.8, for example 0.7, 0.71, 0.73, 0.75, 0.78 or 0.8, etc.
Preferably, the volume ratio of the first chromatographic unit to the fermentation unit is 1 (6-10), e.g. 1:6, 1:6.2, 1:6.5, 1:6.8, 1:7, 1:7.2, 1:7.5, 1:7.8 or 1:8, etc., preferably 1 (8-10).
Preferably, the second chromatography unit is filled with an anion exchange resin, preferably ZGD630 anion exchange resin.
Preferably, the volume filling factor of the anion exchange resin in the second chromatography unit is 0.7 to 0.8, for example 0.7, 0.71, 0.73, 0.75, 0.78 or 0.8, etc.
Preferably, the volume ratio of the second chromatography unit to the fermentation unit is 1 (6-10), e.g. 1:6, 1:6.2, 1:6.5, 1:6.8, 1:7, 1:7.2, 1:7.5, 1:7.8 or 1:8, etc., preferably 1 (8-10).
In a second aspect, a method for co-producing propionic acid and salts thereof and succinic acid and salts thereof using the system according to one of the objects, comprising the steps of:
(1) And (3) strain fermentation: culturing propionibacterium acidogenesis in a fermentation medium, and transferring the culture medium into the fermentation tank for fermentation;
(2) Membrane separation: introducing the fermentation liquor into the membrane separation unit, filtering out fermentation clear liquid, and returning the thallus cells to the fermentation liquor in the step (1);
(3) Acidification of fermentation broth: acidifying the fermentation supernatant obtained in step (2) in the acidification unit to a pH of 3.0 or less, for example 3.0, 2.5, 2.0, 1.5 or 1.2, etc., to obtain an acidified fermentation supernatant;
(4) Adsorption of succinic acid: loading the acidified fermentation liquor obtained in the step (3) to the first chromatographic unit, and adsorbing succinic acid to a saturated state, thereby obtaining a first permeate;
(5) Adsorption of propionic acid: loading the first permeate obtained in the step (4) to the second chromatographic unit, and adsorbing propionic acid to obtain a second permeate; the second permeate returns to the fermentation broth in the step (1);
(6) Collection of propionic acid and salts thereof, succinic acid and salts thereof: eluting the first chromatographic unit with alkali liquor to obtain succinate, or eluting the first chromatographic unit with acid liquor to obtain succinic acid;
eluting the second chromatographic unit with alkali liquor to obtain propionate, or eluting the second chromatographic unit with acid liquor to obtain propionic acid.
The fermentation medium in the step (1) comprises the following components: glucose 50-60 g/L, such as 50g/L, 52g/L, 55g/L, 58g/L or 60g/L, etc., corn steep liquor 41-51 g/L, such as 41g/L, 43g/L, 45g/L, 48g/L or 51g/L, etc., potassium dihydrogen phosphate 3.2-4.6 g/L, such as 3.2g/L, 3.5g/L, 3.8g/L, 4g/L, 4.3g/L or 4.6g/L, etc.
Preferably, the temperature of the propionibacterium acidophilus in step (1) is 28 to 37 ℃, for example 28 ℃, 28.2 ℃, 28.5 ℃, 28.8 ℃, 29 ℃, 29.3 ℃, 29.6 ℃, 29.9 ℃, 30 ℃, 30.2 ℃, 30.5 ℃, 30.8 ℃, 31 ℃, 31.2 ℃, 31.5 ℃, 31.8 ℃, 32 ℃, 33 ℃, 34 ℃, 35 ℃, 36 ℃, 37 ℃ or the like, preferably 30 to 32 ℃.
Preferably, the time for culturing the propionibacterium acidophilus in the step (1) is 36 to 54 hours, for example 36 hours, 37 hours, 38 hours, 39 hours, 40 hours, 41 hours, 42 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 hours, 49 hours, 50 hours, 52 hours or 54 hours, etc., preferably 44 to 50 hours.
Preferably, the filling factor of the fermentation broth in step (1) is 50-80%, such as 50%, 55%, 60%, 65%, 70%, 75% or 80%, etc., preferably 65-75%, and the filling factor is 50-80% so that the volume of the fermenter can be fully utilized and no material leakage occurs during the fermentation process.
Preferably, the temperature of the fermentation broth of step (1) is 28 to 37 ℃, e.g. 28 ℃, 28.2 ℃, 28.5 ℃, 28.8 ℃, 29 ℃, 29.3 ℃, 29.6 ℃, 29.9 ℃, 30 ℃, 30.2 ℃, 30.5 ℃, 30.8 ℃, 31 ℃, 31.2 ℃, 31.5 ℃, 31.8 ℃, 32 ℃, 33 ℃, 34 ℃, 35 ℃, 36 ℃, 37 ℃ or the like, preferably 30 to 32 ℃.
Preferably, the fermentation in step (1) is carried out for a period of 78 to 96 hours, for example 78 hours, 79 hours, 80 hours, 81 hours, 82 hours, 82.5 hours, 82.8 hours, 83 hours, 83.2 hours, 83.5 hours, 83.8 hours, 84 hours, 84.2 hours, 84.5 hours, 84.8 hours, 85 hours, 85.2 hours, 85.5 hours, 85.8 hours, 86 hours, 87 hours, 88 hours, 89 hours, 90 hours, 92 hours, 94 hours or 96 hours, etc., preferably 82 to 86 hours.
Preferably, the pH of the fermentation broth in step (1) is from 6.8 to 7.2, for example, from 6.8, 6.9, 7.0, 7.1 or 7.2, etc., so that the cells remain well grown in a neutral environment.
Preferably, the method for maintaining the pH of the fermentation broth in step (1) comprises: adding ammonia water, naOH and Na 2 CO 3 Or NaHCO 3 Any one or a combination of at least two, wherein typical but non-limiting combinations are: combination of ammonia water and NaOH, na 2 CO 3 And NaHCO 3 Combinations of (2) ammonia and Na 2 CO 3 NaOH and Na 2 CO 3 Is a combination of (a) and (b).
Preferably, the stirring is performed during the fermentation of the strain in step (1).
Preferably, the stirring speed is 50-200 r/min, such as 50r/min, 55r/min, 60r/min, 65r/min, 70r/min, 75r/min, 80r/min, 85r/min, 90r/min, 95r/min, 100r/min, 110r/min, 120r/min, 150r/min, 180r/min or 200r/min, etc., preferably 50-100 r/min.
Preferably, nutrients are supplied during the fermentation of step (1).
Preferably, the nutrients of step (1) include any one or a combination of at least two of glucose, corn steep liquor, yeast extract or peptone, wherein typical but non-limiting combinations are: a combination of glucose and corn steep liquor, a combination of yeast extract and peptone, a combination of glucose and peptone, and a combination of glucose and yeast extract.
Preferably, the means for supplying glucose comprises: 20 to 50wt% of a glucose solution, for example, 20wt%, 21wt%, 22wt%, 25wt%, 28wt%, 30wt%, 32wt%, 35wt%, 38wt%, 40wt%, 42wt%, 45wt%, 48wt% or 50wt% of a glucose solution or the like, preferably 45 to 50wt% of a glucose solution, is added.
Preferably, the nutrient substance supply method in step (1) includes feeding.
Preferably, the nutrient substance in step (1) is supplied in an amount that ensures a carbon equivalent of 4g/L or more in the fermentation broth, for example, the nutrient substance has a carbon equivalent of 4g/L, 5g/L, 6g/L, 7g/L, 8g/L, 9g/L, 10g/L, 12g/L, 14g/L, 15g/L, 18g/L, 20g/L, etc., and the carbon equivalent is preferably 4 to 8g/L. A carbon equivalent of less than 4g/L or more than 8g/L affects the growth of cells, which results in a shortage of carbon sources, and further affects the fermentation efficiency, which results in a glucose effect.
The concentration of propionic acid in the fermentation broth introduced into the membrane separation unit in the step (2) of the present invention is 35g/L or more, for example, 35g/L, 35.5g/L, 36g/L, 36.5g/L, 37g/L, 38g/L, 39g/L, 40g/L, or the like.
Preferably, step (2) is preceded by: and cleaning the membrane separation unit.
Preferably, the cleaning comprises: washing with 0.5-1 wt% sodium bisulphite solution in a membrane separation unit for 40-60 min, such as 40min, 42min, 45min, 48min, 50min, 52min, 55min, 58min or 60min, etc. For example, the concentration of the sodium bisulfite solution is 0.5wt%, 0.6wt%, 0.7wt%, 0.8wt%, 0.9wt% or 1wt%, etc.
Preferably, the pressure during the membrane separation in step (2) is in the range of 0 to 1.5MPa, for example 0MPa, 0.1MPa, 0.2MPa, 0.3MPa, 0.4MPa, 0.5MPa, 0.6MPa, 0.8MPa, 1MPa, 1.2MPa or 1.5MPa, etc., preferably 0 to 0.5MPa.
Preferably, the flow rate of the fermentation broth during the membrane separation in step (2) is 5 to 20L/min, for example 5L/min, 6L/min, 7L/min, 8L/min, 9L/min, 10L/min, 12L/min, 15L/min, 18L/min or 20L/min, etc., preferably 5 to 10L/min.
Along with the continuous exudation of the fermentation clear liquid from the membrane separation unit, the original fermentation liquid is concentrated gradually, so that the on-line separation of the bacterial cells is realized, the inhibition of propionic acid on fermentation is relieved, and the fermentation filtrate can be directly loaded to a subsequent acidification unit and a chromatographic separation unit.
The acidification in the step (3) of the invention specifically comprises the following steps: and (3) loading the fermentation clear liquid obtained in the step (2) into an acidification unit for cation exchange.
Preferably, the loading speed in step (3) is 1-3 BV/h, for example 1BV/h, 1.2BV/h, 1.5BV/h, 1.8BV/h, 2BV/h, 2.1BV/h, 2.2BV/h, 2.3BV/h, 2.4BV/h, 2.5BV/h, 2.6BV/h, 2.7BV/h, 2.8BV/h, 2.9BV/h or 3BV/h, etc., preferably 2-3 BV/h.
Preferably, the loading volume in step (3) is less than or equal to 3BV, for example, 1BV, 1.2BV, 1.5BV, 1.8BV, 2BV, 2.1BV, 2.2BV, 2.3BV, 2.4BV, 2.5BV, 2.6BV, 2.7BV, 2.8BV, 2.9BV, or 3BV, and the like, preferably 2 to 3BV.
The exchange between the feed liquid and the resin requires a certain time, and when the sample loading speed is higher than 3BV/h, the retention time of the feed liquid in the chromatographic unit is too short, and the exchange is incomplete; on the other hand, when the loading speed is lower than 2BV/h, the operation time is prolonged although the exchange is complete, and therefore, the loading speed is preferably 2-3 BV/h.
Likewise, when the loading volume is less than 2BV, the acidification capacity of the resin is insufficient to fully embody, and when the loading volume is greater than 3BV, the expected acidification effect is not achieved, so that the loading volume is 2-3 BV.
After the step (3), the method preferably further comprises:
step (3'): and recycling the acidification unit after regeneration treatment.
Preferably, the step (3') is specifically: and (3) acid liquor is loaded to the acidification unit for regeneration treatment.
Preferably, the acid solution for the regeneration treatment in the step (3') comprises an HCl solution, preferably an HCl solution having a concentration of 1 to 3mol/L, for example, an HCl solution having a concentration of 1 to 2mol/L, 1.2mol/L, 1.5mol/L, 1.8mol/L, 2mol/L, 2.5mol/L or 3mol/L, more preferably an HCl solution having a concentration of 1 to 2 mol/L.
Preferably, the loading rate of the regeneration treatment in step (3') is 1 to 3BV/h, for example 1BV/h, 1.2BV/h, 1.5BV/h, 1.8BV/h, 2BV/h, 2.1BV/h, 2.2BV/h, 2.3BV/h, 2.4BV/h, 2.5BV/h, 2.6BV/h, 2.7BV/h, 2.8BV/h, 2.9BV/h or 3BV/h, etc., preferably 2 to 3BV/h.
The sample loading speed in the step (4) is 1-3 BV/h, for example 1BV/h, 1.2BV/h, 1.5BV/h, 1.8BV/h, 2BV/h, 2.1BV/h, 2.2BV/h, 2.3BV/h, 2.4BV/h, 2.5BV/h, 2.6BV/h, 2.7BV/h, 2.8BV/h, 2.9BV/h or 3BV/h, etc., preferably 2-3 BV/h.
Preferably, the loading volume in step (4) is 6.5BV or more, for example 6.5BV, 7BV, 8BV, 9BV, 10BV, 12BV or 15BV, etc., preferably 10-12 BV.
When the sample loading speed is more than 3BV/h, incomplete exchange of the feed liquid and the resin can be caused; conversely, when the loading rate is lower than 2BV/h, the operation time is prolonged although the exchange is complete. The permeate of the first chromatographic unit can be divided into 3 cases, when the loading volume is smaller than 6.5BV, succinic acid does not penetrate, the resin can not be saturated in the adsorption of the succinate, and propionic acid is adsorbed; when the loading volume is 6.5-10 BV, the succinic acid part penetrates, but the adsorption of the resin on the succinate is not saturated yet; when the loading volume is 10 BV-12 BV, the adsorption of the resin to the succinic acid is saturated, at the moment, the part containing the succinic acid in the first penetrating fluid is additionally collected, the succinic acid is re-loaded to the first chromatographic unit after the first chromatographic unit is regenerated, the succinic acid can only enter the second chromatographic unit after being adsorbed, and when the loading volume is more than 12BV, the adsorption amount of the resin to the succinic acid is not increased any more, and the complex process of repeated absorption of the succinic acid is also increased.
The sample loading speed in the step (5) is 1-3 BV/h, for example, 1BV/h, 1.2BV/h, 1.5BV/h, 1.8BV/h, 2BV/h, 2.1BV/h, 2.2BV/h, 2.3BV/h, 2.4BV/h, 2.5BV/h, 2.6BV/h, 2.7BV/h, 2.8BV/h, 2.9BV/h or 3BV/h, etc., preferably 2-3 BV/h.
Preferably, the loading volume in step (5) is less than or equal to 3BV, such as 1BV, 1.2BV, 1.5BV, 1.8BV, 2BV, 2.1BV, 2.2BV, 2.3BV, 2.4BV, 2.5BV, 2.6BV, 2.7BV, 2.8BV, 2.9BV or 3BV, and the like, preferably 2 to 3BV.
Preferably, in the step (5), when the concentration of the acrylic acid in the second permeate is less than 10g/L, the adsorption is stopped. At this time, the feedback inhibition of propionic acid to fermentation in the fermentation unit is obviously weakened, namely, the inhibition of propionic acid to fermentation is relieved.
When the sample loading speed is more than 3BV/h, incomplete exchange of the feed liquid and the resin can be caused; conversely, when the loading rate is lower than 2BV/h, the operation time is prolonged although the exchange is complete. When the loading volume is more than 3BV, the propionic acid starts to pass through the column, and the loading volume is preferably 2-3 BV in order to improve the yield of the propionic acid.
The alkali solution in the step (6) of the present invention comprises a NaOH solution, preferably a NaOH solution having a concentration of 1 to 4mol/L, for example, a NaOH solution having a concentration of 1mol/L, 1.5mol/L, 2mol/L, 2.5mol/L, 3mol/L, 3.5mol/L or 4mol/L, etc., more preferably a NaOH solution having a concentration of 2.5 to 3 mol/L.
Preferably, the rate of elution of the lye of step (6) is 0.3 to 0.6BV/h, e.g.0.3 BV/h, 0.35BV/h, 0.4BV/h, 0.45BV/h, 0.5BV/h, 0.55BV/h or 0.6BV/h etc., preferably 0.4 to 0.5BV/h.
Preferably, the acid solution of step (6) comprises H 2 SO 4 The solution preferably has a concentration of H of 1 to 4mol/L 2 SO 4 Solutions, e.g. H at a concentration of 1mol/L, 1.5mol/L, 2mol/L, 2.5mol/L, 3mol/L, 3.5mol/L, 3.6mol/L, 3.7mol/L, 3.8mol/L, 3.9mol/L or 4mol/L 2 SO 4 The solution is more preferably H with a concentration of 3.5 to 4mol/L 2 SO 4 A solution.
Preferably, the acid liquid in step (6) is eluted at a rate of 0.3 to 0.8BV/h, for example 0.3BV/h, 0.4BV/h, 0.5BV/h, 0.55BV/h, 0.6BV/h, 0.65BV/h, 0.70BV/h, 0.75BV/h or 0.8BV/h, etc., preferably 0.5 to 0.8BV/h.
Preferably, the step (6) further comprises:
step (6'): and recycling the first chromatographic unit and/or the second chromatographic unit after regeneration treatment.
Preferably, step (6') specifically comprises: and loading alkali liquor into the first chromatographic unit and/or the second chromatographic unit, and performing regeneration treatment.
Preferably, the alkali solution for the regeneration treatment in the step (6') comprises a NaOH solution, preferably a NaOH solution having a concentration of 1 to 4mol/L, for example, a NaOH solution having a concentration of 1mol/L, 1.5mol/L, 2mol/L, 2.5mol/L, 3mol/L, 3.5mol/L, 3.6mol/L, 3.7mol/L, 3.8mol/L, 3.9mol/L or 4mol/L, more preferably a NaOH solution having a concentration of 1 to 2 mol/L.
Preferably, the loading rate of the regeneration treatment in step (6') is 1 to 3BV/h, for example 1BV/h, 1.2BV/h, 1.5BV/h, 1.8BV/h, 2BV/h, 2.1BV/h, 2.2BV/h, 2.3BV/h, 2.4BV/h, 2.5BV/h, 2.6BV/h, 2.7BV/h, 2.8BV/h, 2.9BV/h or 3BV/h, etc., preferably 2 to 3BV/h.
As a preferable technical scheme of the invention, the method for co-producing propionic acid and salts thereof and succinic acid and salts thereof comprises the following steps:
(1) And (3) strain fermentation: culturing propionibacterium acidophilum in a fermentation culture medium at 28-37 ℃ for 36-54 h, wherein the culture medium comprises the following components: glucose 50-60 g/L, corn steep liquor 41-51 g/L, phosphoric acid3.2-4.6 g/L of potassium dihydrogen; then transferring the mixture into a fermentation tank, stirring and fermenting the mixture for 78 to 96 hours at a speed of 50 to 200r/min at a temperature of between 28 and 37 ℃, and feeding nutrient substances with carbon equivalent of more than or equal to 4g/L and adding ammonia water, naOH and Na during the fermentation 2 CO 3 Or NaHCO 3 Any one or a combination of at least two of the above materials maintains the pH of the fermentation broth between 6.8 and 7.2.
(2) Membrane separation: cleaning the membrane separation unit by 0.5-1wt% sodium bisulphite solution in the membrane separation unit for 40-60 min, introducing the fermentation liquor into the membrane separation unit according to the flow rate of 5-20L/min when the propionic acid concentration in the fermentation liquor in the step (1) is more than or equal to 35g/L, filtering out fermentation clear liquid under the pressure of 0-1.5 Mpa, and returning the bacterial cells to the fermentation liquor in the step (1);
(3) Acidification of fermentation broth: loading the fermentation clear liquid obtained in the step (2) into an acidification unit according to the speed of 1-3 BV/h for cation exchange, wherein the loading volume is less than or equal to 3BV, and the pH value is less than or equal to 3.0, so as to obtain the acidification fermentation clear liquid;
(3') introducing acid liquor into the acidification unit for regeneration treatment;
(4) Adsorption of succinic acid: loading the acidified fermentation clear liquid obtained in the step (3) to a first chromatographic unit according to the speed of 1-3 BV/h, wherein the loading volume is more than or equal to 6.5BV, and adsorbing succinic acid to a saturated state, so as to obtain a first permeate;
(5) Adsorption of propionic acid: loading the first permeate obtained in the step (4) to a second chromatographic unit according to the speed of 1-3 BV/h, wherein the loading volume is less than or equal to 3BV, adsorbing propionic acid, and stopping adsorbing when the concentration of the propionic acid in the second permeate is less than 10g/L, so as to obtain a second permeate; the second permeate returns to the fermentation broth in the step (1);
(6) Collection of propionic acid (salt) and succinic acid (salt): eluting the first chromatographic unit with alkali liquor according to the speed of 0.3-0.6 BV/h to obtain succinate, or eluting the first chromatographic unit with acid liquor according to the speed of 0.3-0.6 BV/h to obtain succinic acid;
Eluting the second chromatographic unit with alkali liquor according to the speed of 0.3-0.6 BV/h to obtain propionate, or eluting the second chromatographic unit with acid liquor according to the speed of 0.3-0.6 BV/h to obtain propionic acid.
(6') feeding lye to the first and/or second chromatographic unit for regeneration treatment.
Compared with the prior art, the invention has at least the following beneficial effects:
(1) The invention realizes the co-production of propionic acid and succinic acid through the integration of a fermentation technology, a membrane separation technology and a chromatography technology, and improves the fermentation efficiency and the substrate conversion rate, wherein the yield of the propionic acid is more than 70.34g/L, and the production efficiency is more than 0.440 g/(L.h); the yield of the succinic acid is more than 23.5g/L, and the production efficiency is more than 0.147 g/(L.h);
(2) Compared with the traditional batch fermentation process, the process realizes the on-line separation of fermentation products and thalli, effectively solves the feedback effect of propionic acid on fermentation, and overcomes the defects of low filling efficiency of an expanded bed and difficult mass production;
(3) The process method realizes the secondary utilization of the fermentation waste liquid, and effectively reduces the discharge of the fermentation waste water.
Drawings
FIG. 1 is a front view of a production apparatus used in the examples;
description of the reference numerals
11: a fermentation tank; 12: a nutrient inlet; 13: a stirrer; 21: a membrane separator; 31: acidifying the column; 41: a first chromatographic column; 51: and a second chromatographic column.
Detailed Description
For a better description of the present invention, it is to be understood that the following are exemplary but non-limiting examples of the present invention, which should be construed as merely aiding in the understanding of the present invention and are not to be construed as limiting the invention in any way.
Example 1
A system for co-producing propionic acid and salts thereof and succinic acid and salts thereof, as shown in FIG. 1, comprises a fermentation tank 11, a membrane separator 21, an acidification column 31, a first chromatographic column 41 and a second chromatographic column 51 which are connected in sequence, wherein the fermentation tank 11 is provided with a nutrient inlet 12 and a stirrer 13.
The method for co-producing sodium propionate and sodium succinate by adopting the system shown in figure 1 comprises the following steps:
1) And (3) strain fermentation: inoculating propionibacterium acidophilus stored in glycerol pipe or inclined plane in shake flask fermentation medium, and culturing at 28deg.C for 36 hr, wherein the medium comprises: glucose 60g/L, corn steep liquor 41g/L, and potassium dihydrogen phosphate 4.6g/L; then transferring the mixture into a 5L fermentation tank 11, stirring and fermenting at a speed of 50r/min for 96 hours at a temperature of 28 ℃, wherein the filling coefficient of the fermentation liquid is 50%, adding corn steep liquor with carbon equivalent of 10g/L in a flowing way during the fermentation period, adding ammonia water to maintain the pH value of the fermentation liquid between 6.8 and 7.2, and periodically sampling and measuring the OD600 value of thalli and the concentration of succinic acid and propionic acid in the fermentation liquid;
2) Membrane separation: the pipes of the membrane separator 21 were cleaned by circulating a 0.5wt% sodium bisulphite solution in the membrane separator 21 for 40min, and the solution was charged into a filter having a pore size of 0.1 μm and a filtration area of 0.12m 2 Is a roll film core. When the propionic acid concentration in the fermentation liquid in the step 1) reaches 35g/L, the fermentation tank 11 is connected with the membrane separation 21, the operation is started, the pressure is regulated to 0Mpa, the flow is 5L/min, the fermentation liquid enters the membrane separator 21 for circulation, and the flux per unit area of the membrane core is 20L/(h.m) 2 ) Filtering out fermentation clear liquid, and returning the somatic cells to the fermentation liquid in the step 1);
3) Acidification of fermentation broth: filling 1.5L of H-type cation exchange resin into an acidification column 31, loading the fermentation clear liquid obtained in the step 2) into the acidification column 31 for cation exchange at a speed of 1BV/H, and collecting column penetrating liquid to obtain the acidification fermentation clear liquid, wherein the loading volume is 3BV, and the pH is less than or equal to 3.0;
3') loading HCl solution with the concentration of 1mol/L to an acidification column 31 according to the speed of 1BV/h for regeneration treatment;
4) Adsorption of succinic acid: filling 0.6L of anion exchange resin ZGD630 into a first chromatographic column 41, loading the acidified fermentation liquor obtained in the step 3) into the first chromatographic column 41 at the speed of 1BV/h, and adsorbing succinic acid with the loading volume of 6.5BV to obtain a first permeate;
5) Adsorption of propionic acid: filling 0.8L of anion exchange resin ZGD630 into the second chromatographic column 51, loading the first permeate obtained in the step 4) into the second chromatographic column 51 according to the speed of 1BV/h, wherein the loading volume is equal to 3BV, adsorbing propionic acid, and stopping adsorbing when the concentration of the propionic acid in the second permeate is less than 10g/L, so as to obtain the second permeate; the second permeate returns to the fermentation liquor in the step 1);
6) Collecting sodium propionate and sodium succinate: the first column 41 and the second column 51 were eluted with 1mol/L NaOH solution at a rate of 0.3BV/h to obtain sodium succinate and sodium propionate, respectively.
Example 2
A method for co-producing propionic acid (salt) and succinic acid (salt), which comprises the following steps:
1) And (3) strain fermentation: inoculating propionibacterium acidophilus stored in glycerol pipe or inclined plane in shake flask fermentation medium, and culturing at 37deg.C for 54 hr, wherein the medium comprises: glucose 60g/L, corn steep liquor 41g/L, and potassium dihydrogen phosphate 4.6g/L; then transferring the mixture into a 20L fermentation tank, stirring and fermenting at a speed of 200r/min for 78 hours at 37 ℃, wherein the filling coefficient of the fermentation liquid is 80%, adding peptone with carbon equivalent of 3.5g/L and sodium carbonate into the fermentation tank during the fermentation period to maintain the pH value of the fermentation liquid between 6.8 and 7.2, and periodically sampling and measuring the OD600 value of thalli and the concentration of succinic acid and propionic acid in the fermentation liquid;
2) Membrane separation: cleaning each pipeline of the membrane separator by circulating 1wt% sodium bisulphite solution in the membrane separator for 60min, and loading into a membrane separator with a pore diameter of 0.22 μm and a filtering area of 0.12m 2 Is a roll film core. When the propionic acid concentration in the fermentation liquid in the step 1) reaches 35g/L, the fermentation tank is connected with the membrane in a separated way, the fermentation tank is started to operate, the pressure is regulated to be 1.5Mpa, the flow is 20L/min, the fermentation liquid enters a membrane separator for circulation, and the flux per unit area of the membrane core is 30L/(h.m) 2 ) Filtering out fermentation clear liquid, and returning the somatic cells to the fermentation liquid in the step 1);
3) Acidification of fermentation broth: filling 2L of H-type cation exchange resin into an acidification column, loading the fermentation clear liquid obtained in the step 2) into the acidification column according to the speed of 3BV/H for cation exchange, and collecting the column penetrating liquid as the acidification fermentation clear liquid, wherein the loading volume is 1.0BV until the pH is less than or equal to 3.0;
3') loading HCl solution with the concentration of 1mol/L to an acidification column according to the speed of 3BV/h, and carrying out regeneration treatment;
4) Adsorption of succinic acid: filling 0.8L of anion exchange resin ZGD630 into a first chromatographic column, loading the acidified fermentation liquor obtained in the step 3) into the first chromatographic column according to the speed of 3BV/h, and adsorbing succinic acid to a saturated state by the loading volume of 8BV, thereby obtaining a first permeate;
5) Adsorption of propionic acid: filling 1.0L of anion exchange resin ZGD630 into a second chromatographic column, loading the first permeate obtained in the step 4) into the second chromatographic column according to the speed of 3BV/h, wherein the loading volume is equal to 1.5BV, adsorbing propionic acid, and stopping adsorbing when the concentration of the propionic acid in the second permeate is less than 10g/L, so as to obtain the second permeate; the second permeate returns to the fermentation liquor in the step 1);
6) Collecting sodium propionate and sodium succinate: eluting the first chromatographic column and the second chromatographic column with 4mol/L NaOH solution at a rate of 0.8BV/h to obtain sodium succinate and sodium propionate respectively;
6') 1mol/L NaOH solution is loaded to the first chromatographic column and the second chromatographic column according to the speed of 3BV/h for regeneration treatment.
Example 3
The method for co-producing the propionic acid and the sodium succinate comprises the following steps:
1) And (3) strain fermentation: inoculating propionibacterium acidophilus stored in glycerol pipe or inclined plane in shake flask fermentation medium, and culturing at 30deg.C for 44 hr, wherein the medium comprises: glucose 60g/L, corn steep liquor 41g/L, and potassium dihydrogen phosphate 4.6g/L; then transferring into 7L fermenter, stirring at 30deg.C for 86 hr/min, fermenting with 70% of filling factor of fermentation broth, adding 20wt% glucose solution with carbon equivalent of 4g/L and adding NaHCO during fermentation 3 Maintaining the pH of the fermentation liquor between 6.8 and 7.2, and periodically sampling and measuring the OD600 value of the thalli and the concentration of succinic acid and propionic acid in the fermentation liquor;
2) Membrane separation: the tubes of the membrane separator were circulated with a 0.8wt% sodium bisulfite solution in the membrane separator for 55 minutesCleaning, and loading into a filter with pore diameter of 0.22 μm and filtering area of 0.12m 2 Is a roll film core. When the propionic acid concentration in the fermentation liquid in the step 1) reaches 35g/L, the fermentation tank is connected with the membrane in a separated way, the fermentation tank is started to operate, the pressure is regulated to be 0.5Mpa, the flow is 10L/min, the fermentation liquid enters a membrane separator for circulation, and the flux per unit area of the membrane core is 28L/(h.m) 2 ) Filtering out fermentation clear liquid, and returning the somatic cells to the fermentation liquid in the step 1);
3) Acidification of fermentation broth: filling 1.6L of H-type cation exchange resin into an acidification column, loading the fermentation clear liquid obtained in the step 2) into the acidification column for cation exchange according to the speed of 2BV/H, loading the sample volume of 2BV until the pH value is less than or equal to 3.0, and collecting the column penetrating fluid to obtain the acidification fermentation clear liquid;
3') loading HCl solution with the concentration of 1mol/L to an acidification column according to the speed of 3BV/h, and carrying out regeneration treatment;
4) Adsorption of succinic acid: filling 0.7L of anion exchange resin ZGD630 into a first chromatographic column, loading the acidified fermentation liquor obtained in the step 3) into the first chromatographic column according to the speed of 2BV/h, and adsorbing succinic acid to a saturated state by the loading volume of 10BV, thereby obtaining a first permeate;
5) Adsorption of propionic acid: filling 0.9L of anion exchange resin ZGD630 into a second chromatographic column, loading the first permeate obtained in the step 4) into the second chromatographic column according to the speed of 2BV/h, wherein the loading volume is equal to 2BV, adsorbing propionic acid, and stopping adsorbing when the concentration of the propionic acid in the second permeate is less than 10g/L, so as to obtain the second permeate; the second permeate returns to the fermentation liquor in the step 1);
6) Collection of propionic acid and sodium succinate: eluting the first chromatographic column with 2.5mol/L NaOH solution at a rate of 0.4BV/h to obtain sodium succinate; with 3.5mol/L H 2 SO 4 Eluting the second chromatographic column by the solution to obtain propionic acid;
6') feeding 1mol/L NaOH solution to the first chromatographic column and the second chromatographic column according to the speed of 2BV/h for regeneration treatment.
Example 4
The method for co-producing the propionic acid and the sodium succinate comprises the following steps:
1) And (3) strain fermentation: inoculating propionibacterium acidophilus stored in glycerol pipe or inclined plane in shake flask fermentation medium, and culturing at 32deg.C for 50h, wherein the medium comprises: glucose 60g/L, corn steep liquor 41g/L, and potassium dihydrogen phosphate 4.6g/L; then transferring the mixture into a 10L fermentation tank, stirring and fermenting at the speed of 100r/min for 82 hours at the temperature of 32 ℃, wherein the filling coefficient of the fermentation liquid is 60 percent, 50 weight percent glucose solution with the carbon equivalent of 8g/L is fed in the fermentation period, ammonia water is added in the fermentation period to maintain the pH value of the fermentation liquid between 6.8 and 7.2, and the OD600 value of thalli and the concentration of succinic acid and propionic acid in the fermentation liquid are periodically sampled and measured;
2) Membrane separation: cleaning each pipeline of the membrane separator by circulating sodium bisulphite solution with 0.6wt% in the membrane separator for 45min, and loading into a membrane separator with a pore diameter of 0.22 μm and a filtering area of 0.12m 2 Is a roll film core. When the propionic acid concentration in the fermentation liquid in the step 1) reaches 35g/L, the fermentation tank is connected with the membrane in a separated way, the fermentation tank is started to operate, the pressure is regulated to be 0.5Mpa, the flow is 10L/min, the fermentation liquid enters a membrane separator for circulation, and the flux per unit area of the membrane core is 22L/(h.m) 2 ) Filtering out fermentation clear liquid, and returning the somatic cells to the fermentation liquid in the step 1);
3) Acidification of fermentation broth: filling 1.8L of H-type cation exchange resin into an acidification column, loading the fermentation clear liquid obtained in the step 2) into the acidification column for cation exchange according to the speed of 3BV/H, loading the fermentation clear liquid with the loading volume of 3BV until the pH value is less than or equal to 3.0, and collecting the column penetrating fluid to obtain the acidification fermentation clear liquid;
3') loading the HCl solution with the concentration of 2mol/L to an acidification column according to the speed of 1BV/h, and carrying out regeneration treatment;
4) Adsorption of succinic acid: filling 0.75L of anion exchange resin ZGD630 into a first chromatographic column, loading the acidified fermentation liquor obtained in the step 3) into the first chromatographic column according to the speed of 3BV/h, wherein the loading volume is equal to 10BV, and adsorbing succinic acid to a saturated state, thereby obtaining a first permeate;
5) Adsorption of propionic acid: filling 1L of anion exchange resin ZGD630 into a second chromatographic column, loading the first permeate obtained in the step 4) into the second chromatographic column according to the speed of 3BV/h, wherein the loading volume is equal to 3BV, adsorbing propionic acid, and stopping adsorbing when the concentration of the propionic acid in the second permeate is less than 10g/L, so as to obtain the second permeate; the second permeate returns to the fermentation liquor in the step 1);
6) Collection of propionic acid and sodium succinate: eluting the first chromatographic column with 3mol/L NaOH solution at a rate of 0.5BV/h to obtain sodium succinate; by 4mol/L H 2 SO 4 Eluting the second chromatographic column by the solution to obtain propionic acid;
6') feeding 2mol/L NaOH solution to the first chromatographic column and the second chromatographic column according to the speed of 3BV/h for regeneration treatment.
Example 5
The method for co-producing the propionic acid and the sodium succinate comprises the following steps:
1) And (3) strain fermentation: inoculating propionibacterium acidophilus stored in glycerol pipe or inclined plane in shake flask fermentation medium, and culturing at 30deg.C for 48 hr, wherein the medium comprises: glucose 60g/L, corn steep liquor 41g/L, and potassium dihydrogen phosphate 4.6g/L; then transferring the mixture into a 7L fermentation tank, stirring and fermenting at the speed of 80r/min for 84 hours at the temperature of 30 ℃, wherein the filling coefficient of the fermentation liquid is 65%, adding a 50wt% glucose solution with the carbon equivalent of 6g/L into the fermentation tank, adding ammonia water into the fermentation tank to maintain the pH value of the fermentation liquid between 6.8 and 7.2, and periodically sampling and measuring the OD600 value of thalli and the concentration of succinic acid and propionic acid in the fermentation liquid;
2) Membrane separation: cleaning each pipeline of the membrane separator by circulating 0.7wt% sodium bisulphite solution in the membrane separator for 50min, and loading into a membrane separator with a pore diameter of 0.22 μm and a filtering area of 0.12m 2 Is a roll film core. When the propionic acid concentration in the fermentation liquid in the step 1) reaches 35g/L, the fermentation tank is connected with the membrane in a separated way, the fermentation tank is started to operate, the pressure is regulated to be 0.3Mpa, the flow is 8L/min, the fermentation liquid enters a membrane separator for circulation, and the flux per unit area of the membrane core is 25L/(h.m) 2 ) Filtering out fermentation clear liquid, and returning the somatic cells to the fermentation liquid in the step 1);
3) Acidification of fermentation broth: filling 1.75L of H-type cation exchange resin into an acidification column, loading the fermentation clear liquid obtained in the step 2) into the acidification column for cation exchange according to the speed of 2.5BV/H, and collecting column penetrating liquid to obtain the acidification fermentation clear liquid, wherein the loading volume is 2.5BV until the pH is less than or equal to 3.0;
3') loading HCl solution with the concentration of 1.5mol/L to an acidification column according to the speed of 2.5BV/h, and carrying out regeneration treatment;
4) Adsorption of succinic acid: filling 0.6L of anion exchange resin ZGD630 into a first chromatographic column, loading the acidified fermentation liquor obtained in the step 3) into the first chromatographic column according to the speed of 2.5BV/h, wherein the loading volume is equal to 10BV, and adsorbing succinic acid to a saturated state, thereby obtaining a first permeate;
5) Adsorption of propionic acid: filling 1L of anion exchange resin ZGD630 into a second chromatographic column, loading the second permeate obtained in the step 4) into the second chromatographic column according to the speed of 2.5BV/h, wherein the loading volume is equal to 2.5BV, adsorbing propionic acid, and stopping adsorbing when the concentration of the propionic acid in the first permeate is less than 10g/L, so as to obtain the second permeate; the second permeate returns to the fermentation liquor in the step 1);
6) Collection of propionic acid and sodium succinate: eluting the first chromatographic column with 2.8mol/L NaOH solution at a rate of 0.45BV/h to obtain sodium succinate; with 3.8mol/L H 2 SO 4 Eluting the second chromatographic column by the solution to obtain propionic acid;
6') 1.5mol/L NaOH solution is loaded to the first chromatographic column and the second chromatographic column according to the speed of 2.5BV/h for regeneration treatment.
Comparative example 1
The only difference from example 5 is that: the porous fiber bed reactor constructed by the university of Nanjing industrial Uyang Kai is used for timely separating propionic acid generated in the fermentation process (Journal of Biotechnology,2012, 164:202-210), step 2) is omitted, fermentation liquid is separated after fermentation is carried out according to batches, and propionic acid and succinic acid are respectively extracted.
Comparative example 2
The only difference from example 5 is that: the expanded bed in-situ adsorption proposed by the researchers in the Proc of China process institute Wang Yunshan at Bioresource Technology,2012,104:652-659 is used for replacement, the step 2) is omitted, fermentation liquor is separated after fermentation is carried out according to batches, and propionic acid and succinic acid are respectively extracted.
It should be noted that the process of examples 1 to 5 of the present invention describes only a single cycle of continuous fermentation, after one cycle is completed, the collected column-penetrating liquid from which propionic acid is removed is returned to the fermenter, and nutrient substances are added, the fermentation of step 1) is continued by using the concentrated liquid obtained by membrane separation as seed liquid, and then the operations of cell on-line separation, acidification of fermentation clear liquid, adsorption of succinic acid, adsorption of propionic acid, etc. are continued according to the operations of steps 2) to 6). Only 160 hours are needed per cycle. For convenience of comparison, the productivity and production efficiency of examples 1 to 5 at a production time of 160 hours are collated with comparative example in Table 1. Wherein sodium succinate is converted into equimolar succinic acid for metering, and sodium propionate is converted into equimolar propionic acid for metering.
TABLE 1
As can be seen from Table 1, the yields of propionic acid and succinic acid in example 5 were increased by 56.61% and 36.2% respectively compared with comparative example 1 or 2, and the production efficiencies of propionic acid and succinic acid in example 5 were increased by 56.3% and 35.7% respectively compared with comparative example 1 or 2. Therefore, the invention not only realizes the co-production of the propionic acid and the succinic acid, but also greatly improves the fermentation efficiency and the substrate conversion rate compared with the traditional batch fermentation process, is hopefully applied to the industrialized co-production of the propionic acid and the salt thereof and the succinic acid and the salt thereof, and reduces the industrialization difficulty and the cost.
The applicant states that the detailed structural features of the present invention are described by the above embodiments, but the present invention is not limited to the above detailed structural features, i.e. it does not mean that the present invention must be implemented depending on the above detailed structural features. It should be apparent to those skilled in the art that any modifications of the present invention, equivalent substitutions of selected components of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope of the present invention and the scope of the disclosure.
The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited to the specific details of the above embodiments, and various simple modifications can be made to the technical solution of the present invention within the scope of the technical concept of the present invention, and all the simple modifications belong to the protection scope of the present invention.
In addition, the specific features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, various possible combinations are not described further.
Moreover, any combination of the various embodiments of the invention can be made without departing from the spirit of the invention, which should also be considered as disclosed herein.

Claims (84)

1. A method for co-producing propionic acid and salts thereof and succinic acid and salts thereof, the method comprising the steps of:
(1) And (3) strain fermentation: culturing propionibacterium acidogenesis in a fermentation medium, and transferring the culture medium into the fermentation tank for fermentation;
(2) Membrane separation: introducing the fermentation liquor into the membrane separation unit, filtering out fermentation clear liquid, and returning the thallus cells to the fermentation liquor in the step (1);
(3) Acidification of fermentation broth: acidifying the fermentation clear liquid obtained in the step (2) in the acidification unit until the pH value is less than or equal to 3.0 to obtain acidified fermentation clear liquid;
(4) Adsorption of succinic acid: loading the acidified fermentation liquor obtained in the step (3) to the first chromatographic unit, and adsorbing succinic acid, thereby obtaining a first permeate;
(5) Adsorption of propionic acid: loading the first permeate obtained in the step (4) to the second chromatographic unit, and adsorbing propionic acid to obtain a second permeate; the second permeate returns to the fermentation broth in the step (1);
(6) Collection of propionic acid and salts thereof, succinic acid and salts thereof: eluting the first chromatographic unit with alkali liquor to obtain succinate, or eluting the first chromatographic unit with acid liquor to obtain succinic acid;
eluting the second chromatographic unit with alkali liquor to obtain propionate, or eluting the second chromatographic unit with acid liquor to obtain propionic acid;
The system used by the method comprises a fermentation unit, a membrane separation unit, an acidification unit, a first chromatography unit and a second chromatography unit which are connected in sequence;
a first loop is arranged between the membrane separation unit and the fermentation unit;
a second loop is arranged between the second chromatography unit and the fermentation unit;
the material of the filtering membrane in the membrane separation unit comprises polyethersulfone, the pore diameter of the filtering membrane in the membrane separation unit is 0.1-0.22 mu m, and the unit area flux of the filtering membrane in the membrane separation unit is 20-30L/(h.m) 2 ) The filtering area of the filtering membrane in the membrane separation unit is more than or equal to 0.12m 2 。
2. The method of claim 1, wherein the fermentation unit comprises a fermentor.
3. The method of claim 1, wherein the acidification unit is filled with a cation exchange resin.
4. The method of claim 1, wherein the acidification unit is filled with an H-type cation exchange resin.
5. The method of claim 1, wherein the cation exchange resin in the acidification unit has a volume packing factor of from 0.7 to 0.8.
6. The method of claim 1, wherein the volume ratio of the acidification unit to the fermentation unit is 1 (2-4).
7. The method of claim 1, wherein the first chromatography unit is packed with an anion exchange resin.
8. The method of claim 7, wherein the anion exchange resin is ZGD630 anion exchange resin.
9. The method of claim 1, wherein the anion exchange resin in the first chromatography unit has a volume packing factor of 0.7 to 0.8.
10. The method of claim 1, wherein the volume ratio of the first chromatographic unit to the fermentation unit is 1 (6-10).
11. The method of claim 10, wherein the volume ratio of the first chromatographic unit to the fermentation unit is 1 (8-10).
12. The method of claim 1, wherein the second chromatography unit is packed with an anion exchange resin.
13. The method of claim 12, wherein the anion exchange resin is ZGD630 anion exchange resin.
14. The method of claim 1, wherein the volume filling factor of the anion exchange resin in the second chromatography unit is between 0.7 and 0.8.
15. The method of claim 1, wherein the volume ratio of the second chromatographic unit to the fermentation unit is 1 (6-10).
16. The method of claim 15, wherein the volume ratio of the second chromatographic unit to the fermentation unit is 1 (8-10).
17. The method of claim 1, wherein the medium of step (1) comprises: glucose 50-60 g/L, corn steep liquor 41-51 g/L, and potassium dihydrogen phosphate 3.2-4.6 g/L.
18. The method according to claim 1, wherein the temperature of the cultured propionibacterium acidophilus in step (1) is 28 to 37 ℃.
19. The method of claim 18, wherein the temperature of the cultured propionibacterium acidophilus is between 30 and 32 ℃.
20. The method according to claim 1, wherein the time for culturing propionibacterium acidophilus in step (1) is 36 to 54 hours.
21. The method of claim 20, wherein the propionibacterium acidophilus is cultured for a period of 44-50 hours.
22. The method of claim 1, wherein the fermentation broth of step (1) has a fill factor of 50 to 80%.
23. The method of claim 22, wherein the fermentation broth has a fill factor of 65 to 75%.
24. The method of claim 1, wherein the fermentation broth of step (1) has a temperature of 28 to 37 ℃.
25. The method of claim 24, wherein the fermentation broth is at a temperature of 30 to 32 ℃.
26. The method of claim 1, wherein the fermentation in step (1) is performed for a period of 78 to 96 hours.
27. The method of claim 26, wherein the fermentation is for a period of time ranging from 82 to 86 hours.
28. The method of claim 1, wherein the pH of the fermentation broth is maintained between 6.8 and 7.2 during the fermentation of step (1).
29. The method of claim 28, wherein the means for maintaining the pH of the fermentation broth comprises: adding ammonia water, naOH and Na 2 CO 3 Or NaHCO 3 Any one or a combination of at least two of these.
30. The method of claim 1, wherein the step (1) is performed with stirring during fermentation of the species.
31. The method of claim 30, wherein the stirring is at a rate of 50 to 200r/min.
32. The method of claim 31, wherein the stirring is at a rate of 50 to 100r/min.
33. The method of claim 1, wherein nutrients are supplied during the fermentation of step (1).
34. The method of claim 33, wherein the nutrient comprises any one or a combination of at least two of glucose, corn steep liquor, yeast extract, or peptone.
35. The method of claim 33, wherein the means for supplying glucose comprises: adding 20-50 wt% glucose solution.
36. The method of claim 35, wherein the glucose solution has a concentration of 45 to 50wt%.
37. The method of claim 33, wherein the nutrient is supplied by a method comprising feeding.
38. The method of claim 33, wherein the nutrient is provided in an amount that ensures a carbon equivalent of 4g/L or more in the fermentation broth.
39. The method of claim 38, wherein the carbon equivalent is 4 to 8g/L.
40. The method of claim 1, wherein the propionic acid concentration in the fermentation broth introduced into the membrane separation unit in step (2) is 35g/L or more.
41. The method of claim 1, further comprising, prior to step (2): and cleaning the membrane separation unit.
42. The method of claim 41, wherein the cleaning comprises: cleaning in a membrane separation unit for 40-60 min by using 0.5-1 wt% sodium bisulfite solution.
43. The method of claim 1, wherein the pressure during the membrane separation in step (2) is from 0 to 1.5MPa.
44. The method of claim 43, wherein the pressure during the membrane separation is between 0 and 0.5MPa.
45. The method of claim 1, wherein the flow rate of the fermentation broth during the membrane separation in step (2) is 5 to 20L/min.
46. The method of claim 45, wherein the flow rate of the fermentation broth during the membrane separation is between 5 and 10L/min.
47. The method according to claim 1, wherein the acidification of step (3) comprises in particular: and (3) loading the fermentation clear liquid obtained in the step (2) into an acidification unit for cation exchange.
48. The method of claim 1, wherein the loading rate in step (3) is 1-3 BV/h.
49. The method of claim 48, wherein the loading rate is 2-3 BV/h.
50. The method of claim 17, wherein the loading volume of step (3) is 3BV or less.
51. The method of claim 50, wherein the loading volume is 2-3 BV.
52. The method of claim 17, wherein after step (3), further comprising:
step (3'): and recycling the acidification unit after regeneration treatment.
53. The method of claim 52, wherein step (3') is specifically: and (3) acid liquor is loaded to the acidification unit for regeneration treatment.
54. The method of claim 53, wherein the acid solution for the regeneration treatment of step (3') comprises an HCl solution.
55. The method of claim 54, wherein the concentration of the HCl solution is between 1 and 3mol/L.
56. The method of claim 55, wherein the concentration of the HCl solution is 1-2 mol/L of HCl solution.
57. The method of claim 52, wherein the loading rate of the regeneration process in step (3') is 1-3 BV/h.
58. The method of claim 57, wherein the loading rate of the regeneration process is 2-3 BV/h.
59. The method of claim 1, wherein the loading rate in step (4) is 1-3 BV/h.
60. The method of claim 59, wherein the loading rate is 2-3 BV/h.
61. The method of claim 1, wherein the loading volume of step (4) is 6.5BV or greater.
62. The method of claim 61, wherein the loading volume is between 10 and 12BV.
63. The method of claim 1, wherein the loading rate in step (5) is 1-3 BV/h.
64. The method of claim 63, wherein the loading rate is 2 to 3BV/h.
65. The method of claim 1, wherein the loading volume of step (5) is 3BV or less.
66. The method of claim 65, wherein the loading volume is 2-3 BV.
67. The method of claim 1, wherein in step (5), adsorption is stopped when the concentration of propionic acid in the second permeate is less than 10 g/L.
68. The method of claim 1, wherein the lye of step (6) comprises a NaOH solution.
69. The method of claim 68, wherein the concentration of the NaOH solution is 1 to 4mol/L.
70. The method of claim 69, wherein the NaOH solution has a concentration of 2.5 to 3mol/.
71. The method according to claim 1, wherein the alkaline solution in step (6) is eluted at a rate of 0.3 to 0.6BV/h.
72. The method of claim 71, wherein the lye elution is at a rate of 0.4 to 0.5BV/h.
73. The method of claim 1, wherein the acid solution of step (6) comprises H 2 SO 4 A solution.
74. The method of claim 73, wherein the acid is H at a concentration of 1-4 mol/L 2 SO 4 A solution.
75. The method of claim 73, wherein the acid is H at a concentration of 3.5 to 4mol/L 2 SO 4 A solution.
76. The method of claim 1, wherein the acid in step (6) is eluted at a rate of 0.3 to 0.8BV/h.
77. The method of claim 76, wherein the acid solution elutes at a rate of 0.5 to 0.8BV/h.
78. The method of claim 1, wherein step (6) further comprises, after:
step (6'): and recycling the first chromatographic unit and/or the second chromatographic unit after regeneration treatment.
79. The method of claim 78, wherein step (6') comprises: and loading alkali liquor into the first chromatographic unit and/or the second chromatographic unit, and performing regeneration treatment.
80. The method of claim 78, wherein the lye for the regeneration treatment of step (6') comprises a NaOH solution.
81. The method according to claim 80, wherein the lye for regeneration treatment is a NaOH solution having a concentration of 1 to 4 mol/L.
82. The method according to claim 81, wherein the alkali solution for the regeneration treatment is a NaOH solution having a concentration of 1 to 2 mol/L.
83. The method of claim 78, wherein the loading rate of the regeneration process of step (6') is from 1 to 3BV/h.
84. The method of claim 83, wherein the regeneration process has a loading rate of 2 to 3BV/h.
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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101215583A (en) * 2008-01-18 2008-07-09 南京工业大学 Method for preparing succinic acid by coupling fermentation and membrane separation unit
CN101679893A (en) * 2006-12-28 2010-03-24 安塔克有限公司 Material and/or fuel produced from biomass
CN101967498A (en) * 2009-07-27 2011-02-09 中国科学院过程工程研究所 Method for producing biological organic acid by using dynamic expanding adsorption reactor system
CN103667373A (en) * 2012-09-05 2014-03-26 深圳市绿微康生物工程有限公司 Method for generating propionic acid by microbial fermentation and co-generating succinic acid and salt thereof
WO2014089812A1 (en) * 2012-12-13 2014-06-19 南京工业大学 Method for producing ethanol by coupling immobilization bed fermentation with separation
CN105385712A (en) * 2015-11-03 2016-03-09 沈阳科纳提克生物科技有限公司 Fermentation and production method for propionic acid
CN206916118U (en) * 2017-05-09 2018-01-23 中国科学院过程工程研究所 A kind of coproduction propionic acid and its salt and the system of succinic acid and its salt

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101679893A (en) * 2006-12-28 2010-03-24 安塔克有限公司 Material and/or fuel produced from biomass
CN101215583A (en) * 2008-01-18 2008-07-09 南京工业大学 Method for preparing succinic acid by coupling fermentation and membrane separation unit
CN101967498A (en) * 2009-07-27 2011-02-09 中国科学院过程工程研究所 Method for producing biological organic acid by using dynamic expanding adsorption reactor system
CN103667373A (en) * 2012-09-05 2014-03-26 深圳市绿微康生物工程有限公司 Method for generating propionic acid by microbial fermentation and co-generating succinic acid and salt thereof
WO2014089812A1 (en) * 2012-12-13 2014-06-19 南京工业大学 Method for producing ethanol by coupling immobilization bed fermentation with separation
CN105385712A (en) * 2015-11-03 2016-03-09 沈阳科纳提克生物科技有限公司 Fermentation and production method for propionic acid
CN206916118U (en) * 2017-05-09 2018-01-23 中国科学院过程工程研究所 A kind of coproduction propionic acid and its salt and the system of succinic acid and its salt

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
崔晓蓬."用离子交换层析技术提取发酵液中丙酸".《中国优秀硕士学位论文全文数据库工程科技Ⅰ辑》.2017,(第3期),第1-27页. *
崔晓蓬等."用离子交换层析技术提取发酵液中丙酸".《过程工程学报》.2016,第16卷(第3期),第1-6页. *

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