CN112176005B - Production method of calcium propionate - Google Patents
Production method of calcium propionate Download PDFInfo
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- CN112176005B CN112176005B CN202011167788.7A CN202011167788A CN112176005B CN 112176005 B CN112176005 B CN 112176005B CN 202011167788 A CN202011167788 A CN 202011167788A CN 112176005 B CN112176005 B CN 112176005B
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- 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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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Abstract
The invention discloses a production method of calcium propionate, belonging to the field of biochemical engineering. Compared with the traditional fermentation method for preparing calcium propionate, the method disclosed by the invention can ensure the optimal acid yield in the growth, production and aging processes of the fermentation bacteria by controlling the pH value of the fermentation bacteria in the fermentation process; meanwhile, after the fermented mixture is filtered and concentrated, a calcium source is added for neutralization reaction, and compared with the method of directly adding the calcium source into a culture medium for one-step culture production of calcium propionate, the method can effectively reduce the influence on the purity of a final product caused by excessive or insufficient reaction raw materials. The method has the advantages of simple operation steps, low requirement on production instruments, low production cost, high production yield, low water content of the finally obtained calcium propionate product and high content of effective components.
Description
Technical Field
The invention relates to the field of biochemical engineering, and in particular relates to a production method of calcium propionate.
Background
Calcium propionate is used as a common food preservative, has excellent preservative and fresh-keeping functions, and is good in dissolving performance and high in effective calcium content in the food production process, so that the calcium propionate is more beneficial to human bodies compared with the traditional preservative. At present, the main preparation process of calcium propionate mainly combines and prepares propionic acid and a calcium source directly, and the process has high production consumption and has the risk of environmental pollution; on the other hand, a preparation process for preparing calcium propionate by a biological fermentation method has been reported, and the method prepares the propionic acid by a fermentation method, and then adds a calcium source to prepare the calcium propionate. However, the propionic acid generated by fermentation in the method is easy to inhibit subsequent acid production of the fermentation bacteria and synthesis of calcium propionate, so that the production efficiency cannot be ensured; the calcium propionate product prepared by this method also has the disadvantage of low purity.
Disclosure of Invention
Based on the defects of the prior art, the invention aims to provide the production method of the calcium propionate, which has the characteristics of high preparation efficiency, simple operation steps and high product purity.
In order to achieve the purpose, the invention adopts the technical scheme that:
a method for producing calcium propionate, comprising the steps of:
(1) culturing an acid-producing strain at normal temperature, inoculating the acid-producing strain to a fermentation culture medium, and fermenting to obtain a fermentation mixture A; the fermentation process realizes the control of the generation rate of propionic acid by adjusting the pH value to 7-2.5;
(2) and (2) sterilizing, filtering, nano-filtering and concentrating the fermentation mixture A obtained in the step (1), adding a calcium hydroxide aqueous solution for reaction, and concentrating and drying the mixed solution after the reaction is completed to obtain solid calcium propionate.
Compared with the traditional fermentation method for preparing calcium propionate, the production method of calcium propionate provided by the invention can ensure the optimal acid yield in the growth, production and aging processes of the zymocyte by controlling the pH value of the zymocyte in the fermentation process; meanwhile, the addition amount of the calcium source is determined by the pH value during the neutralization reaction, and compared with the method of directly adding the calcium source into the culture medium to produce calcium propionate through one-step culture, the method can effectively reduce the influence on the purity of the final product caused by excessive or insufficient reaction raw materials. The method has the advantages of simple operation steps, low requirements on production instruments, low production cost and high production yield, and the finally obtained calcium propionate product has low water content and effective components of more than or equal to 90 percent.
Preferably, the acid producing strain is lactobacillus. The lactobacillus is taken as a bacterium source which is widely distributed and has aerobic and anaerobic characteristics, the propagation speed is high, the growth cost is low, meanwhile, the secretion of the lactobacillus can inhibit the growth of other strains to a certain extent, and the purity of propionic acid generated by fermentation can be effectively guaranteed.
Preferably, the culture medium cultured in the step (1) comprises glucose and peptone, and the inoculation amount of the acid-producing strain in the culture medium is 10-20% of the mass of the culture medium. The culture of the acid-producing strain is carried out through the culture medium and the inoculation amount, so that the propagation speed and the growth activity of the acid-producing strain can be ensured.
Preferably, the temperature of the culture in the step (1) is 25-35 ℃, the pressure is 0.02-0.04 MPa, and the time is 22-26 h. The acid-producing strain can keep higher reproductive activity under the culture condition, and meanwhile, mutual inhibition among colonies is not caused.
Preferably, the strain is stirred every 6-10 hours in the culture process in the step (1), the stirring time is 4-6 min each time, and the rest time is static culture. The aerobic state of all parts in the culture environment can be ensured by stirring irregularly, and the inhibition of secretion aggregation of acid-producing strains on the propagation of bacterial colonies is avoided. More preferably, the culture process implements on-line pH detection and interval sampling detection to ensure that the acid-base environment in the culture environment is stable and free from foreign bacteria pollution. And inoculating the strain into a fermentation medium for fermentation when the pH value is maintained at 4.5-5.5 and no foreign bacteria are detected.
Preferably, the fermentation medium in the step (1) contains glucose, yeast extract and peptone, the inoculation amount of the acid-producing strain in the fermentation medium is 10-20% of the mass of the fermentation medium, the fermentation period is 240 hours, and the temperature is 25-35 ℃. The fermentation substrate and the fermentation conditions can effectively ensure that acid-producing strains can be sufficiently propagated and produce propionic acid, and the production efficiency of the acid-producing strains cannot be inhibited due to overhigh concentration of the propionic acid.
More preferably, the fermentation of step (1) is divided into four stages according to the fermentation cycle, the four stages controlling the pH of the fermentation medium: the first stage is 0-72 h, and the pH is controlled to be 6.8-7; the second stage is 72-144 h, and the pH is controlled to be 5-6; the third stage is 144-192 hours, and the pH is controlled to be 4-5; the fourth stage is 192-240 hours, and the pH is controlled to be 2.5-3.5. In the whole fermentation period, the pH of each stage is strictly controlled according to the reproduction and propionic acid production rules of acid-producing strains, so that the stable acid production amount of the acid-producing strains in the rapid growth period, the acid production active period and the growth aging period can be effectively ensured, the produced propionic acid is prevented from inhibiting the reproduction of the strains, and the fermentation production efficiency is ensured.
More preferably, the fermentation process is implemented by sampling at intervals, the detected items are the content of various acids (including propionic acid, lactic acid, acetic acid and succinic acid) in the fermentation medium, and the interval time is 24 h. The pH value and the concentration of various acid-producing agents in the fermentation environment can be effectively known through interval sampling detection, so that the pH value can be controlled in a targeted manner.
Preferably, the adjustment of the pH in step (1) is carried out by adding an aqueous glucose solution or an aqueous sodium hydroxide solution. The pH adjustment by the mild additives avoids adverse effects on strain propagation and acid production. More preferably, the mass content of glucose in the glucose aqueous solution is 35-45%, and the mass content of sodium hydroxide in the sodium hydroxide aqueous solution is 20-30%.
Preferably, the sterilization temperature in the step (2) is 120-130 ℃, the pressure is 0.1MPa, and the time is 25-35 min. The acid-producing strains and additional foreign strains can be effectively inactivated through the sterilization process under the optimal conditions, and pollution caused by subsequent production is avoided.
Preferably, the filtering in the step (2) is carried out at a temperature of 60-70 ℃, and the pore diameter of the ceramic membrane is 200-500 nm. The ceramic membrane is used for filtering, so that the unconsumed fermentation medium and the inactivated strains in the fermentation liquor can be effectively filtered, and the purity of the target solid in the solution is higher.
Preferably, the concentration multiple of the nanofiltration concentration in the step (2) is 3-5, and the solid content of the concentrated solution after the nanofiltration concentration is 20-30%. By removing the excessive water in the inactivated and filtered filtrate, the content of the effective components in the solution can be improved, and the purity of the finally prepared calcium propionate product is further improved.
Preferably, the mass content of the calcium hydroxide in the calcium hydroxide aqueous solution in the step (2) is 20-30%. More preferably, the mass content of calcium hydroxide in the calcium hydroxide aqueous solution is 25%. By limiting the content of calcium hydroxide in the solution, the purity of the final product caused by insufficient or excessive raw materials can be avoided, the volume amount of the reaction solution can be kept moderate, and the limitation of the actual production process by a reaction container can be avoided.
The method has the beneficial effects that the pH value in the fermentation environment is strictly controlled in the fermentation process of the acid-producing strain, so that the stable acid-producing rate of the strain can be maintained at each stage of the growth period, and the production efficiency of the strain is ensured; the acid production is filtered and concentrated to react with calcium hydroxide with a characteristic content to finally generate calcium propionate, so that waste caused by too much or too little raw materials can be avoided, and the yield and the purity of a final product can be fully ensured.
Detailed Description
Unless otherwise specified, the raw materials used in the examples of the present invention and comparative examples were commercially available, and the production equipment used was a commercially available common model.
For better illustrating the objects, technical solutions and advantages of the present invention, the present invention will be further described with reference to specific examples, which are intended to be understood in detail, but not intended to limit the present invention.
The culture medium used by the invention is a lactobacillus culture medium produced by Haibo biology Limited company; the fermentation medium used was a sugar fermentation medium produced by Haibo biology, Inc.
Example 1
One embodiment of the method of producing calcium propionate of the present invention.
The production method of the calcium propionate comprises the following steps:
(1) sterilizing a culture medium at 121 ℃ and 0.1MPa for 30min, inoculating lactobacillus strains with 15% of inoculum size, culturing at 30 ℃ and 0.02-0.04 MPa for 24h, stirring the strains every 8h, stirring for 5min each time, standing for culture in the rest time, and detecting the pH of the culture medium and the pollution condition of the mixed bacteria at intervals in a spot inspection mode. When the pH value is maintained at 4.5-5.5 and no mixed bacteria pollution exists, inoculating the mixture into a fermentation tank containing a fermentation culture medium according to the inoculation amount of 15%, and fermenting for 240 hours at 30 ℃ to obtain a fermentation mixture A; the fermentation process is divided into four stages according to the fermentation period, and the four stages are respectively controlled by 25% sodium hydroxide water solution to control the pH value: the first stage is 0-72 h, and the pH is controlled to be 6.8-7; the second stage is 72-144 h, and the pH is controlled to be 5-6; the third stage is 144-192 hours, and the pH is controlled to be 4-5; the fourth stage is 192-240 hours, and the pH is controlled to be 2.5-3.5. During the period, sampling detection is carried out at intervals, the content of various acids (including propionic acid, lactic acid, acetic acid and succinic acid) in the fermentation medium is detected, and the interval time is 24 h.
(2) Sterilizing the fermentation mixture A obtained in the step (1) for 30min at 121 ℃, under 0.1MPa, filtering by a ceramic membrane with 450nm pore diameter at 65 ℃, keeping the pH at 2.5-3, performing nanofiltration and concentration for 4 times until the solid content is 25%, adding 25% of calcium hydroxide aqueous solution at 45 ℃ for reaction, and concentrating and drying the mixed solution after the reaction is completed to obtain solid calcium propionate.
By calculation, 5000kg of lactic acid bacteria residues are generated by the batch of filtration, and the culture condition of the strains is good; 8600kg of calcium hydroxide is used as a raw material to produce 10000kg of solid calcium propionate product, so that the product yield is high; the detection shows that the product has the effective components up to 92%, the water content only 3% and high product purity.
Example 2
One embodiment of the method of producing calcium propionate of the present invention.
The production method of the calcium propionate comprises the following steps:
(1) sterilizing a culture medium at 121 ℃ and 0.1MPa for 30min, inoculating lactobacillus strains with the inoculation amount of 20%, culturing at 25 ℃ and 0.02-0.04 MPa for 22h, stirring the strains every 10h, stirring for 6min every time, standing for culturing at the rest time, and detecting the pH of the culture medium and the pollution condition of the mixed bacteria at intervals in a spot inspection mode. When the pH value is maintained at 4.5-5.5 and no mixed bacteria pollution exists, inoculating the mixture into a fermentation tank containing a fermentation culture medium according to the inoculation amount of 20%, and fermenting for 240 hours at 25 ℃ to obtain a fermentation mixture A; the fermentation process is divided into four stages according to the fermentation period, and the four stages are respectively controlled by 40% glucose water solution to have pH: the first stage is 0-72 h, and the pH is controlled to be 6.8-7; the second stage is 72-144 h, and the pH is controlled to be 5-6; the third stage is 144-192 hours, and the pH is controlled to be 4-5; the fourth stage is 192-240 hours, and the pH is controlled to be 2.5-3.5. During the period, sampling detection is carried out at intervals, the content of various acids (including propionic acid, lactic acid, acetic acid and succinic acid) in the fermentation medium is detected, and the interval time is 24 h.
(2) Sterilizing the fermentation mixture A obtained in the step (1) for 35min at 130 ℃, under 0.1MPa, filtering by a ceramic membrane with the aperture of 250nm at 70 ℃, keeping the pH value at 2.5-3, performing nanofiltration and concentration for 5 times until the solid content is 25%, adding 30% of calcium hydroxide aqueous solution at 45 ℃ for reaction, and concentrating and drying the mixed solution after the reaction is completed to obtain solid calcium propionate.
Through calculation, 5200kg of lactic acid bacteria residue is produced by the batch of filtration, and the culture condition of the strain is good; 8400kg of calcium hydroxide is used as a raw material to produce 9800kg of solid calcium propionate product, and the product yield is high; the detection shows that the product has 90% of effective components, 4% of water content and high product purity.
Example 3
One embodiment of the method of producing calcium propionate of the present invention.
The production method of the calcium propionate comprises the following steps:
(1) sterilizing a culture medium at 121 ℃ and 0.1MPa for 30min, inoculating lactobacillus strains with the inoculation amount of 10%, culturing at 35 ℃ and 0.02-0.04 MPa for 26h, stirring the strains every 6h, stirring for 4min each time, standing for culturing at the rest time, and detecting the pH of the culture medium and the pollution condition of the mixed bacteria at intervals in a spot inspection mode. When the pH value is maintained at 4.5-5.5 and no foreign bacteria pollution exists, inoculating the mixture into a fermentation tank containing a fermentation culture medium according to the inoculation amount of 10%, and fermenting for 240 hours at 35 ℃ to obtain a fermentation mixture A; the fermentation process is divided into four stages according to the fermentation period, and the four stages are respectively controlled by 30% sodium hydroxide aqueous solution to have pH value: the first stage is 0-72 h, and the pH is controlled to be 6.8-7; the second stage is 72-144 h, and the pH is controlled to be 5-6; the third stage is 144-192 hours, and the pH is controlled to be 4-5; the fourth stage is 192-240 hours, and the pH is controlled to be 2.5-3.5. During the period, sampling detection is carried out at intervals, the content of various acids (including propionic acid, lactic acid, acetic acid and succinic acid) in the fermentation medium is detected, and the interval time is 24 h.
(2) Sterilizing the fermentation mixture A obtained in the step (1) for 25min at 120 ℃ and 0.1MPa, filtering the fermentation mixture A by using a ceramic membrane with the aperture of 500nm at 60 ℃, keeping the pH value at 2.5-3, performing nanofiltration and concentration for 3 times until the solid content is 25%, adding a 20% calcium hydroxide aqueous solution at 45 ℃ for reaction, and concentrating and drying the mixed solution after the reaction is completed to obtain solid calcium propionate.
Through calculation, 4800kg of lactic acid bacteria residues are generated by the batch of filtration, and the culture condition of the strains is good; the method can produce 12000kg of solid calcium propionate product by using 8600kg of calcium hydroxide as a raw material, and has high product yield; the detection shows that the product has 90% of effective components, 4% of water content and high product purity.
Example 4
This example differs from example 1 only in that the amount of lactobacillus inoculated in the fermenter according to step (1) was 25%
Through calculation, the batch of the lactic acid bacteria residues are generated by filtering, and the culture condition of the strains is general; the total use of 7900kg of calcium hydroxide as a raw material can produce 9000kg of solid calcium propionate product, and the product yield is low; the detection shows that the product has 87 percent of effective components, 5 percent of water content and higher product purity.
Example 5
This example differs from example 1 only in that the amount of lactobacillus inoculated in the fermenter according to step (1) was 5%
According to calculation, 4000kg of lactic acid bacteria residues are generated by the batch of filtration, and the culture condition of the strains is general; 8500kg of solid calcium propionate product can be produced by using 7600kg of calcium hydroxide as a raw material, and the product yield is low; the detection shows that the product has the effective components up to 88%, the water content of only 5% and high product purity.
Example 6
The difference between this example and example 1 is only that the fermentation process in step (2) is divided into two stages according to the fermentation cycle, and the two stages are respectively controlled in pH by 25% aqueous sodium hydroxide solution: the first stage is 0-144 h, and the pH is controlled to be 7-5; the second stage is 144-240 h, and the pH is controlled to be 5-2.5.
Through calculation, 4500kg of lactic acid bacteria residues are generated by the batch of filtration, 8100kg of solid calcium propionate product can be produced by using 7000kg of calcium hydroxide as a raw material, and the product yield is low; the detection shows that the product has 87 percent of effective components, 5 percent of water content and higher product purity.
Example 7
This example differs from example 1 only in that the fermentation process described in step (2) is controlled in pH by 25% aqueous sodium hydroxide: the first stage is 0-144 h, and the pH is controlled to be 7-2.5.
Through calculation, 3700kg of lactic acid bacteria residues are generated in the batch of filtration, and the culture condition of the strains is poor; 7500kg of solid calcium propionate product can be produced by using 6200kg of calcium hydroxide as a raw material, and the product yield is low; the detection shows that the effective components of the product reach 83%, the water content is only 6%, and the product purity is general.
Example 8
This example differs from example 1 only in that the nanofiltration concentration in step (2) is 2-fold and the solids content after concentration is 50%.
By calculation, 5000kg of lactic acid bacteria residues are generated by the batch of filtration, and the culture condition of the strains is good; 12000kg of solid calcium propionate product can be produced by using 8500kg of calcium hydroxide as a raw material, and the product yield is high; the detection shows that the product has 81% of effective components, 9% of water content and lower product purity.
Example 9
The present example differs from example 1 only in that the ceramic membrane described in step (2) has a pore size of 600 nm.
By calculation, 6000kg of lactic acid bacteria residues are generated by the batch of filtration, and the culture condition of the strains is good; 8800kg of calcium hydroxide is used as raw material to produce 14000kg of solid calcium propionate product, and the product yield is higher; the detection shows that the product has effective components up to 80%, water content of 5% and low product purity.
Comparative example 1
The comparative example differs from example 1 only in that no reagent was added to control the pH during the fermentation in step (1).
Through calculation, 3200kg of lactic acid bacteria residues are generated in the batch of filtration, and the culture condition of the strains is poor; the raw material of 5700kg of calcium hydroxide can be used for producing 7200kg of solid calcium propionate product, and the product yield is low; the detection shows that the product has the effective components up to 88%, the water content of 4% and high product purity.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
Claims (5)
1. A method for producing calcium propionate, comprising the steps of:
(1) culturing acid-producing strains, inoculating the acid-producing strains to a fermentation medium, and fermenting to obtain a fermentation mixture A; the inoculation amount of the acid-producing strain in the fermentation culture medium is 10-20% of the mass of the fermentation culture medium; the fermentation period is 240 hours, and the temperature is 25-35 ℃; the fermentation is divided into four stages according to the fermentation period, and the four stages respectively control the pH of the fermentation medium by adding glucose aqueous solution or sodium hydroxide aqueous solution: the first stage is 0-72 h, and the pH is controlled to be 6.8-7; the second stage is 72-144 h, and the pH is controlled to be 5-6; the third stage is 144-192 hours, and the pH is controlled to be 4-5; the fourth stage is 192-240 hours, and the pH is controlled to be 2.5-3.5; the acid-producing strain is lactobacillus;
(2) sterilizing, filtering and nano-filtering the fermentation mixture A obtained in the step (1), concentrating, adding a calcium hydroxide aqueous solution for reaction, concentrating and drying the mixed solution after the reaction is completed to obtain solid calcium propionate; the filtering temperature is 60-70 ℃, the filtering is carried out by using a ceramic membrane, and the aperture of the ceramic membrane is 200-500 nm; the concentration multiple of nanofiltration concentration is 3-5, and the solid content of the concentrated solution after nanofiltration concentration is 20-30%.
2. The method for producing calcium propionate according to claim 1, wherein the inoculation amount of the acid-producing strain in the culture medium in the step (1) is 10-20% of the mass of the culture medium; the culture temperature is 25-35 ℃, the pressure is 0.02-0.04 MPa, and the time is 22-26 h.
3. The method of claim 1, wherein the fermentation medium of step (1) comprises glucose, yeast extract, and peptone.
4. The method for producing calcium propionate according to claim 1, wherein the glucose aqueous solution contains 35 to 45% by mass of glucose, and the sodium hydroxide aqueous solution contains 20 to 30% by mass of sodium hydroxide.
5. The method for producing calcium propionate according to claim 1, wherein the mass content of calcium hydroxide in the aqueous calcium hydroxide solution in the step (2) is 20-30%.
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