CN109182423B - Method for promoting fermentation of escherichia coli to produce polysialic acid - Google Patents
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Abstract
A method for promoting fermentation of Escherichia coli to produce polysialic acid relates to the field of fermentation engineering. The method for promoting the fermentation production of polysialic acid by escherichia coli comprises the following steps: inoculating escherichia coli to a fermentation medium for culture and fermentation, and separating to obtain polysialic acid; during culture and fermentation, the temperature between 0h and 48h of culture and fermentation is controlled to be 40-42 ℃, and the temperature after 48h of culture and fermentation is controlled to be 36-38 ℃. The method for promoting the fermentation of the escherichia coli to produce the polysialic acid can effectively improve the fermentation level, the synthesis efficiency and the carbon source conversion efficiency of the polysialic acid.
Description
Technical Field
The invention relates to the field of fermentation engineering, and in particular relates to a method for promoting escherichia coli to ferment and produce polysialic acid.
Background
N-acetylneuraminic acid (N-acetylneuraminic acid) Neu5Ac is the first contact site for cell information transmission, and the molecular structure of the N-acetylneuraminic acid is diverse, so that Neu5Ac participates in multiple physiological processes such as cell recognition, signal transduction, tumorigenesis and fertilization, Neu5Ac can also regulate the anti-inflammatory activity of IgG, enhance the immunity of infants, influence the integrity, permeability and activity of nerve cells and promote the development of the brains of infants, and therefore the production of N-acetylneuraminic acid attracts more attention and research.
The current production method of N-acetylneuraminic acid comprises the following steps: natural product extraction, chemical synthesis, biological enzyme catalytic conversion, direct fermentation of genetically engineered bacteria, and polysialic acid fermentation hydrolysis. Because the content of Neu5Ac in most natural products is low and the components are very complex, the extraction of Neu5Ac from the natural products is accompanied by a complex process, the recovery rate is low, and the requirement of large-scale production is difficult to meet. The chemical synthesis method cannot be applied to industrial production due to complicated operations such as group protection and deprotection and the existence of chiral isomeric byproducts. The problems of biological enzyme-catalyzed transformation and genetic engineering bacteria related to transgenosis are difficult to be approved by food-related laws, and the biological enzyme-catalyzed transformation and genetic engineering bacteria can not be applied to the field of infant health. Only the polysialic acid fermentation hydrolysis method is easy to be accepted by consumers because the polysialic acid fermentation hydrolysis method belongs to natural products, and has passed official approval in China, so that the polysialic acid fermentation hydrolysis method has higher industrial value.
Polysialic acid (PSA) is a homopolymer of N-acetylneuraminic acid connected by α -2, 8 and/or α -2, 9 bonds and is a main component of capsular polysaccharide of a few bacteria, in 1957, Barry and Goebel firstly find Polysialic acid in E.coli K235, and then people find Polysialic acid in other strains successively, but the fermentation level, the synthesis efficiency and the carbon source conversion efficiency of the Polysialic acid are lower at present, the yield of the Polysialic acid PSA is only 5-6 g/L when the Polysialic acid is produced by fermentation for 30-40h, so that the fermentation cost is higher, and the application of the N-acetylneuraminic acid is limited.
Disclosure of Invention
The invention aims to provide a method for promoting escherichia coli to ferment and produce polysialic acid, which can effectively improve the fermentation level, the synthesis efficiency and the carbon source conversion efficiency of the polysialic acid.
The technical problem to be solved by the invention is realized by adopting the following technical scheme.
A method for promoting fermentation production of polysialic acid by Escherichia coli, which comprises the following steps: inoculating escherichia coli to a fermentation medium for culture and fermentation, and separating to obtain polysialic acid; during culture and fermentation, the temperature between 0h and 48h of culture and fermentation is controlled to be 40-42 ℃, and the temperature after 48h of culture and fermentation is controlled to be 36-38 ℃.
Further, in the preferred embodiment of the present invention, the initial ventilation amount is controlled to be 0.8-1.2vvm, and the initial rotation speed is 150-; after inoculation, the dissolved oxygen content is adjusted to be more than 3 mg/L.
Further, in the preferred embodiment of the present invention, in the above-mentioned culture fermentation, cetyltrimethylammonium bromide was added to the fermentation medium at 8h from the inoculation of Escherichia coli into the fermentation medium so that the cetyltrimethylammonium bromide reached 150-250 mg/L.
Further, in a preferred embodiment of the present invention, sorbitol is supplied to the fermentation medium at a rate of 1 to 4 g/(L.h) after the carbon source in the fermentation medium is consumed during the above-mentioned culture fermentation.
Further, in a preferred embodiment of the present invention, the pH of the culture medium is controlled to 6.0 to 6.5 by using an aqueous ammonia solution.
Further, in a preferred embodiment of the present invention, the fermentation medium comprises the following components: 15-25g/L sorbitol, 2-4g/L corn steep liquor dry powder, 3-7g/L dipotassium hydrogen phosphate, 0.3-0.8g/L magnesium sulfate, 4-8g/L ammonium chloride, 4-8g/L N-acetylglucosamine, 0.5-2mL/L B vitamin solution and 0.5-2mL/L trace element solution.
Further, in a preferred embodiment of the present invention, the vitamin B solution comprises the following components: VB of 0.3-0.6g/L13-3.5g/L VB5VBH of 0.001-0.01g/L and VB of 0.1-0.2g/L12。
Further, in a preferred embodiment of the present invention, the trace element solution comprises the following components: 1-4g/L of FeSO40.01-0.1g/L of KIO30.5-1g/L of MnCl20.1-0.5g/L of CoCl20.01-0.1g/L of CrCl30.01-0.2g/L of ZnSO40.01-0.05g/L of Na2MoO4And 1-2g/L of H3BO3。
Further, in a preferred embodiment of the present invention, the fermentation medium further comprises 0.1-1mL/L sucrose fatty acid ester.
Further, in a preferred embodiment of the present invention, the preservation number of the escherichia coli is CCTCC NO: and M2018103.
The escherichia coli and the application thereof in fermentation production of polysialic acid have the beneficial effects that: the method for promoting the fermentation of escherichia coli to produce the polysialic acid provided by the embodiment of the invention comprises the following steps: inoculating escherichia coli to a fermentation medium for culture and fermentation, and separating to obtain polysialic acid; during culture and fermentation, the temperature between 0h and 48h of culture and fermentation is controlled to be 40-42 ℃, and the temperature after 48h of culture and fermentation is controlled to be 36-38 ℃. The method for promoting the fermentation of the escherichia coli to produce the polysialic acid can effectively improve the fermentation level, the synthesis efficiency and the carbon source conversion efficiency of the polysialic acid.
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In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
FIG. 1 is a graph showing polysialic acid content and absorbance at different fermentation times for the production of polysialic acid by fermentation of Escherichia coli in example 1 of the present invention;
FIG. 2 is a graph showing polysialic acid content and absorbance at different fermentation times for the production of polysialic acid by fermentation of Escherichia coli in comparative example 1 of the present invention;
FIG. 3 is a graph showing polysialic acid content and absorbance at different fermentation times for the production of polysialic acid by fermentation of Escherichia coli in comparative example 2 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The following is a detailed description of the Escherichia coli of the present invention and its use in fermentative production of polysialic acid.
The invention provides a method for promoting escherichia coli to ferment and produce polysialic acid, which comprises the following steps:
inoculating escherichia coli to a fermentation medium for culture and fermentation, and separating to obtain polysialic acid; during culture and fermentation, the temperature between 0h and 48h of culture and fermentation is controlled to be 40-42 ℃, and the temperature after 48h of culture and fermentation is controlled to be 36-38 ℃. Preferably, the Escherichia coli is cultured to obtain a seed solution, and then the seed solution is inoculated into a fermentation medium for culture.
The application of the escherichia coli in fermentation production of the polysialic acid provided by the embodiment of the invention is to inoculate the escherichia coli to a fermentation culture medium for culture and fermentation to obtain the polysialic acid, and the fermentation temperature is controlled to be 40-42 ℃ between 0-48h of culture and fermentation and 36-38 ℃ after 48h of culture and fermentation by adopting a fermentation culture method controlled by stage temperature change, so that the carbon source transfer rate, the PSA fermentation level and the synthesis efficiency can be effectively improved. Thereby improving the production efficiency of polysialic acid.
In a preferred embodiment of the invention, during the culture fermentation, the initial ventilation amount is controlled to be 0.8-1.2vvm, and the initial rotation speed is 150-; after inoculation, the dissolved oxygen content is adjusted to be more than 3 mg/L. By controlling the ventilation index in the fermentation process after inoculating the escherichia coli, the suitable environment for propagation of the escherichia coli can be ensured, and the production of polysialic acid by fermenting the escherichia coli is promoted.
In a preferred embodiment of the present invention, in the cultivation of the fermentation, cetyltrimethylammonium bromide was added to the fermentation medium at 8h from the inoculation of Escherichia coli into the fermentation medium so that the concentration of cetyltrimethylammonium bromide reached 150-250 mg/L. Cetyl trimethyl ammonium bromide is added in the culture fermentation process, so that the shedding of cell capsules can be promoted, and the production rate of polysialic acid is improved.
In a preferred embodiment of the present invention, sorbitol is supplied to the fermentation medium at a rate of 1 to 4 g/(L.h) after the carbon source in the fermentation medium is consumed during the fermentation. By continuously adding the carbon source into the fermentation medium, the polysialic acid can be continuously generated by fermentation after the propagation of escherichia coli.
In a preferred embodiment of the present invention, the pH is controlled to 6.0 to 6.5 by using an aqueous ammonia solution during the fermentation. The pH is preferably controlled by using an aqueous ammonia solution of 5 to 10% by mass. The growth and propagation of Escherichia coli can be promoted by controlling the pH value of culture fermentation, so that the production rate of polysialic acid can be effectively improved.
In a preferred embodiment of the invention, the fermentation medium comprises the following components: 15-25g/L sorbitol, 2-4g/L corn steep liquor dry powder, 3-7g/L dipotassium hydrogen phosphate, 0.3-0.8g/L magnesium sulfate, 4-8g/L ammonium chloride, 4-8g/L N-acetylglucosamine, 0.5-2mL/L B vitamin solution and 0.5-2mL/L trace element solution. Among them, preferably, the B vitamin solution comprises the following components: VB of 0.3-0.6g/L13-3.5g/L VB5VBH of 0.001-0.01g/L and VB of 0.1-0.2g/L12. More preferably, the trace element solution comprises the following components: 1-4g/L of FeSO40.01-0.1g/L of KIO30.5-1g/L of MnCl20.1-0.5g/L of CoCl20.01-0.1g/L of CrCl30.01-0.2g/L of ZnSO40.01-0.05g/L of Na2MoO4And 1-2g/L of H3BO3. The fermentation medium with proper components is prepared, so that the rapid propagation of escherichia coli can be promoted, and the fermentation of the fermentation medium by the escherichia coli is promoted to produce polysialic acid.
In a preferred embodiment of the invention, the fermentation medium further comprises 0.1-1mL/L sucrose fatty acid ester. The sucrose fatty acid ester is added into the fermentation medium as the defoaming agent, so that the generation of foam can be inhibited, the phenomenon that a large amount of foam generated in the fermentation process influences the fermentation process is avoided, and the stable fermentation is ensured to produce and obtain the polysialic acid.
In a preferred embodiment of the present invention, the Escherichia coli is selected from the group consisting of Escherichia coli having a preservation number of CCTCC NO: e.coli strain M2018103. The Escherichia coli is preserved in China typical culture Collection (CCTCC) of eight Loa Jia mountain in Wuchang district, Wuhan city, Hubei province in 2018 at 3 and 6 months, with the preservation number of CCTCC NO: m2018103; taxonomic nomenclature: escherichia coli CASOV-8.
The features and properties of the present invention are described in further detail below with reference to examples.
Example 1
The embodiment of the invention provides a method for producing polysialic acid by using escherichia coli fermentation, which mainly comprises the following steps:
s11, streaking the strain of the frozen escherichia coli (preservation number CCTCC NO: M2018103) to a solid LB culture medium and culturing at 37 ℃ to obtain a single colony; the used Escherichia coli has been submitted to China Center for Type Culture Collection (CCTCC) for preservation in eight Loa Jia mountain in Wuchang district, Wuhan City, Hubei province in 2018, 3 months and 6 days; wherein, the solid LB culture medium contains the following components: 10g/L of peptone, 5g/L of yeast extract powder, 10g/L of sodium chloride and 20g/L of agar; the pH of the solid LB medium was 7.0.
S12, selecting a single colony, inoculating the single colony into a triangular flask containing 50mL of liquid LB seed culture medium, and culturing overnight at the stirring speed of 200rpm and the temperature of 37 ℃ to obtain a first-stage seed solution; wherein the liquid LB seed culture medium contains the following components: 10g/L of peptone, 5g/L of yeast extract powder and 10g/L of sodium chloride; the pH of the liquid LB seed medium was 7.0.
S13, inoculating the primary seed solution into a triangular flask containing 50mL of liquid LB seed culture medium according to the inoculation amount of 1% by volume, and culturing at 37 ℃ for 6h at the stirring speed of 200rpm to obtain a secondary seed solution.
S14, filling 3L of fermentation medium into a 5L fermentation tank, and inoculating 100mL of secondary seed liquid into the fermentation tank for culture; wherein the fermentation medium contains the following components: 20g/L of sorbitol, 3g/L of corn steep liquor dry powder, 5g/L of dipotassium phosphate, 0.5g/L of magnesium sulfate, 6g/L, N of ammonium chloride, 6g/L, B of acetylglucosamine and 1mL/L of trace element solution. Wherein, the B vitamin solution contains the following components: VB of 0.5g/L13.2g/L VB5VBH of 0.006g/L and VB of 0.15g/L12(ii) a The solution of trace elements containsThe following components: 2g/L of FeSO40.06g/L of KIO30.8g/L of MnCl20.2g/L of CoCl20.08g/L of CrCl30.09g/L of ZnSO40.03g/L of Na2MoO4And 1.75g/L of H3BO3。
S15, in the culture process, controlling the temperature between 0 and 48 hours of culture and fermentation to be 40 ℃, the temperature after 48 hours of culture and fermentation to be 37 ℃, controlling the initial ventilation volume after inoculation to be 1vvm and the initial rotation speed to be 200 rpm; adjusting the dissolved oxygen content DO to 3mg/L after inoculation, adjusting the pH value in a fermentation tank at the position of ammonia water with the mass concentration of 10% in the fermentation process to 6.4, adding hexaalkyltrimethylammonium bromide into a fermentation medium in the 8 th hour of fermentation after inoculation to enable the cetyl trimethylammonium bromide in the fermentation medium to reach 200mg/L, and supplementing sorbitol into the fermentation medium at the rate of 2.5 g/(L.h) after the carbon source in the fermentation medium is consumed.
S16, fermenting and culturing for 72h, then canning and taking the fermentation product, and separating to obtain the polysialic acid.
Example 2
The embodiment of the invention provides a method for producing polysialic acid by using escherichia coli fermentation, which mainly comprises the following steps:
s21, streaking the strain of the frozen escherichia coli (preservation number CCTCC NO: M2018103) to a solid LB culture medium and culturing at 37 ℃ to obtain a single colony; wherein, the solid LB culture medium contains the following components: 10g/L of peptone, 5g/L of yeast extract powder, 10g/L of sodium chloride and 20g/L of agar; the pH of the solid LB medium was 7.0.
S22, selecting a single colony, inoculating the single colony into a triangular flask containing 50mL of liquid LB seed culture medium, and culturing overnight at the stirring speed of 200rpm and the temperature of 37 ℃ to obtain a first-stage seed solution; wherein the liquid LB seed culture medium contains the following components: 10g/L of peptone, 5g/L of yeast extract powder and 10g/L of sodium chloride; the pH of the liquid LB seed medium was 7.0.
S23, inoculating the primary seed solution into a triangular flask containing 50mL of liquid LB seed culture medium according to the inoculation amount of 1% by volume, and culturing at 37 ℃ for 6h at the stirring speed of 200rpm to obtain a secondary seed solution.
S24, filling 3L of fermentation medium into a 5L fermentation tank, and inoculating 100mL of secondary seed liquid into the fermentation tank for culture; wherein the fermentation medium contains the following components: 25g/L sorbitol, 4g/L corn steep liquor dry powder, 6g/L dipotassium phosphate, 0.6g/L magnesium sulfate, 8g/L ammonium chloride L, N-acetylglucosamine, 1.5mL/L sucrose fatty acid ester/L, B vitamin solution and 1.5mL/L trace element solution. Wherein, the B vitamin solution contains the following components: VB of 0.6g/L13.5g/L VB5VBH of 0.008g/L and VB of 0.18g/L12(ii) a The trace element solution contains the following components: 3g/L of FeSO40.08g/L of KIO30.9g/L of MnCl20.3g/L of CoCl20.09g/L of CrCl30.1g/L of ZnSO40.05g/L of Na2MoO4And 2g/L of H3BO3。
S25, controlling the temperature between 0 and 48 hours of culture and fermentation to be 40 ℃, the temperature after 48 hours of culture and fermentation to be 37 ℃, controlling the initial ventilation volume after inoculation to be 1.2vvm and the initial rotating speed to be 200rpm in the culture process; adjusting the dissolved oxygen content DO to 3.5mg/L after inoculation, using ammonia water with the mass concentration of 10% in the fermentation tank during the fermentation process, adjusting the pH value in the fermentation tank to 6.4, and supplementing sorbitol into the fermentation medium at the rate of 3 g/(L.h) after the carbon source in the fermentation medium is consumed.
S26, fermenting and culturing for 72h, then canning and taking the fermentation product, and separating to obtain the polysialic acid.
Example 3
The embodiment of the invention provides a method for producing polysialic acid by using escherichia coli fermentation, which mainly comprises the following steps:
s31, streaking the strain of the frozen escherichia coli (preservation number CCTCC NO: M2018103) to a solid LB culture medium and culturing at 37 ℃ to obtain a single colony; wherein, the solid LB culture medium contains the following components: 10g/L of peptone, 5g/L of yeast extract powder, 10g/L of sodium chloride and 20g/L of agar; the pH of the solid LB medium was 7.0.
S32, selecting a single colony, inoculating the single colony into a triangular flask containing 50mL of liquid LB seed culture medium, and culturing overnight at the stirring speed of 200rpm and the temperature of 37 ℃ to obtain a first-stage seed solution; wherein the liquid LB seed culture medium contains the following components: 10g/L of peptone, 5g/L of yeast extract powder and 10g/L of sodium chloride; the pH of the liquid LB seed medium was 7.0.
S33, inoculating the primary seed solution into a triangular flask containing 50mL of liquid LB seed culture medium according to the inoculation amount of 1% by volume, and culturing at 37 ℃ for 6h at the stirring speed of 200rpm to obtain a secondary seed solution.
S34, filling 3L of fermentation medium into a 5L fermentation tank, and inoculating 100mL of secondary seed liquid into the fermentation tank for culture; wherein the fermentation medium contains the following components: sorbitol 18g/L, corn steep liquor dry powder 2.5g/L, dipotassium phosphate 4g/L, magnesium sulfate 0.4g/L, ammonium chloride 5g/L, N-acetylglucosamine 5g/L, B vitamin solution 1mL/L and trace element solution 1 mL/L. Wherein, the B vitamin solution contains the following components: VB of 0.4g/L13.0g/L VB5VBH of 0.005g/L and VB of 0.12g/L12(ii) a The trace element solution contains the following components: 1.5g/L of FeSO40.05g/L of KIO30.6g/L of MnCl20.15g/L of CoCl20.06g/L CrCl30.075g/L of ZnSO40.02g/L of Na2MoO4And 1.6g/L of H3BO3。
S35, controlling the temperature between 0-48h of culture and fermentation to be 40 ℃, the temperature after 48h of culture and fermentation to be 37 ℃, controlling the initial ventilation volume after inoculation to be 0.8vvm and the initial rotating speed to be 200rpm in the culture process; adjusting the dissolved oxygen content DO to 4mg/L after inoculation, adjusting the pH value in a fermentation tank by using ammonia water with the mass concentration of 10% in the fermentation process to 6.4, and supplementing sorbitol into the fermentation medium at the rate of 2 g/(L.h) after the carbon source in the fermentation medium is consumed.
S36, fermenting and culturing for 72h, then canning and taking the fermentation product, and separating to obtain the polysialic acid.
Comparative example 1
The method for producing polysialic acid by fermentation using E.coli provided in this comparative example was substantially the same as the method for producing polysialic acid by fermentation using E.coli in example 1, except that the culturing process in step S15 of this comparative example was carried out at a temperature of 37 ℃ for 0-48 hours of culture fermentation and at a temperature of 37 ℃ after 48 hours of culture fermentation.
Comparative example 2
The method for producing polysialic acid by fermentation using E.coli provided in this comparative example was substantially the same as the method for producing polysialic acid by fermentation using E.coli in example 1, except that the culturing process in step S15 of this comparative example was carried out at a temperature of 40 ℃ between 0 and 48 hours of culture fermentation and at a temperature of 40 ℃ after 48 hours of culture fermentation.
Sampling and detecting in the fermentation tank culture processes of example 1, comparative example 1 and comparative example 2, specifically, during the culture period, taking fermentation products at different time points (fermentation 12H, 24H, 36H, 48H, 60H and 72H), respectively, and detecting the OD600 (absorbance at 600nm wavelength) and the polysialic acid content, wherein the detection method of the polysialic acid content is as shown in figure 1, figure 2 and figure 3, wherein the fermentation liquid is centrifuged to remove thallus, the clear liquid is diluted, 2mL of the diluted clear liquid is mixed with 0.1mol/L of diluted hydrochloric acid, the mixture is sealed and uniformly mixed in a water bath at 85 ℃ for 3H, and after the reaction is finished, the mixture is cooled, and is detected by using HPLC, a High Performance Liquid Chromatography (HPLC) detection method, Shimadzu Lc-15c, a detection Column Bio-Rad AMINEX HPX 87H Organic Analysis Column (300 × 7.8.8 mm), a Column temperature is 60 ℃, a mobile phase is 5mmol of sulfuric acid, the flow rate is 0.6mL/min, and the wavelength is 210 nm.
As can be seen from FIG. 1, in the method for producing polysialic acid by Escherichia coli fermentation provided in example 1, the final PSA yield is 19.1g/L, the synthesis rate is 3.18 g/L.multidot.12h, and the conversion rate of sorbitol is 10.11%; as can be seen from FIG. 2, in the method for producing polysialic acid by Escherichia coli fermentation provided in comparative example 1, the final PSA yield is 15.3g/L, the synthesis rate is 2.55 g/L.multidot.12h, and the conversion rate of sorbitol is 8.16%; as can be seen from FIG. 3, in the method for producing polysialic acid by fermentation of E.coli as provided in comparative example 2, the final PSA yield was 15.3g/L, the synthesis rate was 2.55 g/L.multidot.12h, and the conversion rate of sorbitol was 8.10%. Therefore, the culture fermentation process with the stage temperature change control provided by the invention can effectively improve the PSA fermentation level, the synthesis efficiency and the carbon source conversion efficiency, and promote the fermentation production efficiency of the polysialic acid.
In conclusion, the method for promoting escherichia coli to ferment and produce polysialic acid provided by the embodiment of the invention can effectively improve the carbon source conversion rate, the PSA fermentation level and the PSA synthesis efficiency, thereby effectively improving the production efficiency of polysialic acid.
The embodiments described above are some, but not all embodiments of the invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Claims (7)
1. A method for promoting fermentation production of polysialic acid by Escherichia coli is characterized by comprising the following steps: inoculating escherichia coli to a fermentation medium for culture and fermentation, and separating to obtain polysialic acid; during the culture and fermentation, the temperature between 0h and 48h of the culture and fermentation is controlled to be 40-42 ℃, and the temperature after 48h of the culture and fermentation is controlled to be 36-38 ℃;
in the culture and fermentation process, adding cetyl trimethyl ammonium bromide into the fermentation culture medium in 8h from the inoculation of the escherichia coli into the fermentation culture medium to ensure that the cetyl trimethyl ammonium bromide reaches 150-250 mg/L;
the fermentation medium comprises the following components: 15-25g/L sorbitol, 2-4g/L corn steep liquor dry powder, 3-7g/L dipotassium hydrogen phosphate, 0.3-0.8g/L magnesium sulfate, 4-8g/L ammonium chloride, 4-8g/L N-acetylglucosamine, 0.5-2mL/L B vitamin solution and 0.5-2mL/L trace element solution;
the preservation number of the escherichia coli is CCTCC NO: m2018103.
2. The method for promoting fermentation production of polysialic acid by Escherichia coli as claimed in claim 1, wherein the initial ventilation amount is controlled to be 0.8-1.2vvm, and the initial rotation speed is controlled to be 150-; after inoculation, the dissolved oxygen content is adjusted to be more than 3 mg/L.
3. The method for promoting fermentative production of polysialic acid by Escherichia coli according to claim 1, wherein sorbitol is supplemented to the fermentation medium at a rate of 1-4 g/(L-h) after the carbon source in the fermentation medium is consumed during the fermentation.
4. The method for promoting fermentation production of polysialic acid by Escherichia coli according to claim 1, wherein the pH is controlled to 6.0-6.5 by using ammonia water solution during the fermentation.
5. The method for promoting escherichia coli fermentation to produce polysialic acid according to claim 1, wherein the B vitamin solution comprises the following components: VB of 0.3-0.6g/L13-3.5g/L VB5VBH of 0.001-0.01g/L and VB of 0.1-0.2g/L12。
6. The method for promoting fermentation production of polysialic acid by escherichia coli according to claim 1, wherein the trace element solution comprises the following components: 1-4g/L of FeSO40.01-0.1g/L of KIO30.5-1g/L of MnCl20.1-0.5g/L of CoCl20.01-0.1g/L of CrCl30.01-0.2g/L of ZnSO40.01-0.05g/L of Na2MoO4And 1-2g/L of H3BO3。
7. The method for promoting fermentation production of polysialic acid by escherichia coli according to claim 1, wherein the fermentation medium further comprises 0.1-1mL/L of sucrose fatty acid ester.
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