CN110964762A - Fermentation process of low-starch-residue xanthan gum product - Google Patents
Fermentation process of low-starch-residue xanthan gum product Download PDFInfo
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Abstract
The invention belongs to the technical field of biological fermentation, and discloses a fermentation process of a low-starch-residue xanthan gum product, wherein a used fermentation medium comprises the following components: glucose, corn starch paste, yeast extract, oleic acid, calcium carbonate, magnesium sulfate heptahydrate, dipotassium hydrogen phosphate, fulvic acid and VB1And eugenol. The fermentation process of the invention reduces the viscosity of the fermentation liquor, improves the fermentation efficiency, does not cause starch residue in the fermentation liquor, and leads the subsequent extraction process to be more thorough.
Description
Technical Field
The invention belongs to the technical field of biological fermentation, and particularly relates to a fermentation process of a low-starch-residue xanthan gum product.
Background
Xanthan gum is an acidic extracellular heteropolysaccharide produced by fermentation of wild Xanthomonas using carbohydrate as a main substrate. Because of the unique structure of the xanthan gum, the xanthan gum has excellent performances such as tackifying synergy, low concentration and high viscosity, pseudoplasticity, high temperature resistance, acid resistance, good dispersion effect, emulsion stability and the like, and is widely applied to more than 20 industries such as oil exploitation, chemical industry, textile, medicine, food, cosmetics and the like. The research of many experts and scholars at home and abroad on the xanthan gum production rate mainly focuses on various aspects such as strain breeding, culture medium, fermentation conditions, genetic engineering, enzyme engineering and the like. The xanthan gum production in China still has some problems at present, such as long thallus growth time, insufficient starch utilization, low product quality grade and the like.
In recent years, research institutes and related enterprises have been devoted to study optimization of xanthan gum fermentation conditions. The addition of sodium and potassium ions to the culture medium can facilitate the transfer of the energy and precursor substance for xanthan biosynthesis, glucose, into the cell and also facilitate the transfer of the precursor substance for xanthan biosynthesis out of the cell. However, in the xanthan biosynthesis process, the substrate metabolism flux, cell membrane permeability, cytochrome P450 enzyme and the like all have an influence on the synthesis of xanthan, but there are few studies on this aspect.
Research shows that under the stress conditions of high salt, high alkali, high temperature, drought and the like, the content of extracellular polysaccharide produced by bacteria is likely to increase, but under the stress conditions, the proliferation activity of the strain is deteriorated, and the level of the finally obtained xanthan gum is not necessarily improved; the literature 'influence of organic solvent stress treatment on extracellular polysaccharide production of a strain Bacillus subtilis OST23a, report of microbiology 2012' shows that the extracellular polysaccharide yield secreted by marine bacteria can be improved by adding 3% of n-hexane stress into a culture solution, but the addition of an excessive organic solvent can cause environmental pollution and the removal of the organic solvent in a fermentation solution is relatively difficult; in addition, whether the normal hexane stress can improve the xanthan gum producing capability of the xanthomonas campestris is still to be researched.
Chinese patent CN109929893A, a fermentation process of low-cost high-quality xanthan gum, comprising the following steps: inoculating xanthomonas campestris into a fermentation tank filled with a fermentation culture medium for fermentation culture, coupling the fermentation tank with a ceramic membrane, fermenting for 48h, separating the fermentation liquor in the fermentation tank by the ceramic membrane to obtain filtrate and concentrated thalli, discharging the filtrate into a feed liquid storage tank, pumping the concentrated thalli back to the fermentation tank, simultaneously supplementing the fermentation tank with the fermentation culture medium, continuing to ferment for 36-48h to obtain the fermentation liquor, separating the fermentation liquor in the fermentation tank by the ceramic membrane to obtain the filtrate and the concentrated thalli, and discharging the filtrate into the feed liquid storage tank. The method improves the total yield of xanthan gum, but improves the consumption of fermentation medium, and increases the enterprise cost. The existing xanthan gum production process also has the following problems: the common process is not specially controlled by people, and the starch residue of the product has uncertainty; in particular, the starch residue requirement of food-grade products is strict, and the qualification rate of finished products is lower. On the basis of the above patent technology, in combination with the problems existing in the prior art, the xanthan gum fermentation conditions are continuously optimized.
Disclosure of Invention
In order to further optimize the xanthan gum fermentation conditions, the invention provides a fermentation process of the xanthan gum product with low starch residue.
The invention is realized by the following technical scheme.
The fermentation process of the low-starch-residue xanthan gum product is characterized in that the fermentation medium used in the fermentation process comprises the following components: glucose, corn starch paste, yeast extract, oleic acid, calcium carbonate, magnesium sulfate heptahydrate, dipotassium hydrogen phosphate, fulvic acid and VB1And eugenol.
Preferably, the components of the fermentation medium are:
60g/L glucose, 120g/L corn starch paste, 10g/L yeast extract, 10g/L oleic acid, 3g/L calcium carbonate, 1g/L magnesium sulfate heptahydrate, 1g/L dipotassium phosphate and 20mg/L, VB fulvic acid120mg/L, 1mg/L of eugenol and pH 7.0-7.2.
Preferably, the components of the fermentation medium are:
60g/L glucose, 100g/L corn starch paste, 10g/L yeast extract, 10g/L oleic acid, 3g/L calcium carbonate, 1g/L magnesium sulfate heptahydrate, 1g/L dipotassium phosphate and 20mg/L, VB fulvic acid120mg/L, 1.5mg/L eugenol and pH 7.0-7.2.
Preferably, the preparation method of the fermentation medium comprises the following steps:
sequentially adding the raw materials into water, stirring, and adjusting pH.
Preferably, the corn starch paste is prepared by enzymatic hydrolysis with amylase.
More preferably, the preparation method of the corn starch paste comprises the following steps: adding water into corn starch to adjust the mass concentration of the starch milk to 50%, adjusting the pH to 5.5 and the temperature to 50 ℃ to prepare high-concentration starch milk; adding alpha-amylase into high-concentration starch milk, heating to 60 ℃, performing enzymolysis for 30-60min under the stirring condition of 100rpm, heating to 100 ℃, inactivating enzyme, and naturally cooling to room temperature to obtain the alpha-amylase. Most preferably, the a-amylase is added in an amount of 10U per gram of corn starch.
Most preferably, the enzymolysis time is 40-50 min.
Most preferably, the enzyme deactivation time is 5 min.
Further, the fermentation process specifically comprises: inoculating the seed liquid of the xanthomonas into a full-automatic fermentation tank filled with a fermentation culture medium for fermentation culture at the fermentation temperature of 30 ℃ for 66-72h to obtain fermentation liquid.
Compared with the prior art, the invention has the advantages that the following aspects are mainly included but not limited:
the medium-temperature a-amylase can hydrolyze α -l and 4-glucoside bonds in starch molecules, so that the DE value of liquefied liquid is increased rapidly, the viscosity of starch paste is decreased rapidly, the viscosity value is decreased to be below 40 mPa.s at the temperature of 60 ℃, so that the viscosity of fermentation liquid is reduced, the fermentation efficiency is improved, starch residue is not caused in the fermentation liquid, and the subsequent extraction process is more thorough.
Drawings
FIG. 1: influence of fermentation time on the amount of produced gum;
FIG. 2: the influence of the addition of eugenol in the fermentation medium on the gum yield.
Detailed Description
Those skilled in the art can modify the process parameters appropriately to achieve the desired results with reference to the disclosure herein. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the products and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations and modifications, or appropriate alterations and combinations, of the products and methods described herein may be made and utilized without departing from the spirit, scope, and spirit of the invention. For a further understanding of the present invention, reference will now be made in detail to the following examples.
Example 1
A fermentation process for a low starch residue xanthan gum product comprising the steps of:
xanthomonas ATCC 17915 seed liquid (3X 10)8CFU/mL) is inoculated into a 500L full-automatic fermentation tank filled with 300L fermentation medium according to the inoculation amount of 10 percent (volume ratio) for fermentation culture at the fermentation temperature of 30 ℃ for 72 hours to obtain fermentation liquid; in the whole fermentation process, defoaming is carried out by feeding foam killer, residual sugar is controlled to be not less than 1% by feeding glucose solution, and the dissolved oxygen level is kept to be 20% by adjusting the stirring speed and the ventilation quantity;
the fermentation medium comprises the following components:
60g/L glucose, 120g/L corn starch paste, 10g/L yeast extract, 10g/L oleic acid, 3g/L calcium carbonate, 1g/L magnesium sulfate heptahydrate, 1g/L dipotassium phosphate and 20mg/L, VB fulvic acid120mg/L, 1mg/L of eugenol and pH 7.0-7.2.
The preparation method of the fermentation medium comprises the following steps:
sequentially adding the raw materials into water, stirring, and adjusting pH.
The preparation method of the corn starch paste comprises the following steps:
adding water into corn starch to adjust the mass concentration of the starch milk to 50%, adjusting the pH to 5.5 and the temperature to 50 ℃ to prepare high-concentration starch milk; adding medium-temperature a-amylase (10000U/g) into high-concentration starch milk, wherein the addition amount of the a-amylase is 10U per gram of starch, heating to 60 ℃, performing enzymolysis for 50min under the stirring condition of 100rpm, heating to 100 ℃, inactivating the enzyme for 5min, and naturally cooling to room temperature to obtain the alpha-amylase. Example 2
A fermentation process for a low starch residue xanthan gum product comprising the steps of:
xanthomonas ATCC 17915 seed liquid (4X 10)8CFU/mL) is inoculated into a 500L full-automatic fermentation tank filled with 300L fermentation medium according to the inoculation amount of 10 percent (volume ratio) for fermentation culture at the fermentation temperature of 30 ℃ for 66 hours to obtain fermentation liquid; in the whole fermentation process, defoaming is carried out by feeding foam killer, residual sugar is controlled to be not less than 1% by feeding glucose solution, and the dissolved oxygen level is kept to be 20% by adjusting the stirring speed and the ventilation quantity;
the fermentation medium comprises the following components:
60g/L glucose, 100g/L corn starch paste, 10g/L yeast extract, 10g/L oleic acid, 3g/L calcium carbonate, 1g/L magnesium sulfate heptahydrate, 1g/L dipotassium phosphate and 20mg/L, VB fulvic acid120mg/L, 1.5mg/L eugenol and pH 7.0-7.2.
The preparation method of the fermentation medium comprises the following steps:
sequentially adding the raw materials into water, stirring, and adjusting pH.
The preparation method of the corn starch paste comprises the following steps:
adding water into corn starch to adjust the mass concentration of the starch milk to 50%, adjusting the pH to 5.5 and the temperature to 50 ℃ to prepare high-concentration starch milk; adding medium-temperature a-amylase (10000U/g) into high-concentration starch milk, wherein the addition amount of the a-amylase is 10U per gram of starch, heating to 60 ℃, performing enzymolysis for 40min under the stirring condition of 100rpm, heating to 100 ℃, inactivating the enzyme for 5min, and naturally cooling to room temperature to obtain the alpha-amylase. Comparative example 1
A fermentation process for a low starch residue xanthan gum product comprising the steps of:
yellow sheetSeed liquid of Cytospora ATCC 17915 (3X 10)8CFU/mL) is inoculated into a 500L full-automatic fermentation tank filled with 300L fermentation medium according to the inoculation amount of 10 percent (volume ratio) for fermentation culture at the fermentation temperature of 30 ℃ for 72 hours to obtain fermentation liquid; in the whole fermentation process, defoaming is carried out by feeding foam killer, residual sugar is controlled to be not less than 1% by feeding glucose solution, and the dissolved oxygen level is kept to be 20% by adjusting the stirring speed and the ventilation quantity;
the fermentation medium comprises the following components:
60g/L glucose, 60g/L corn starch, 10g/L yeast extract, 10g/L oleic acid, 3g/L calcium carbonate, 1g/L magnesium sulfate heptahydrate, 1g/L dipotassium phosphate and 20mg/L, VB fulvic acid120mg/L，pH 7.0-7.2。
The preparation method of the fermentation medium comprises the following steps:
sequentially adding the raw materials into water, stirring, and adjusting pH.
Comparative example 2
A fermentation process for a low starch residue xanthan gum product comprising the steps of:
xanthomonas ATCC 17915 seed liquid (3X 10)8CFU/mL) is inoculated into a 500L full-automatic fermentation tank filled with 300L fermentation medium according to the inoculation amount of 10 percent (volume ratio) for fermentation culture at the fermentation temperature of 30 ℃ for 72 hours to obtain fermentation liquid; in the whole fermentation process, defoaming is carried out by feeding foam killer, residual sugar is controlled to be not less than 1% by feeding glucose solution, and the dissolved oxygen level is kept to be 20% by adjusting the stirring speed and the ventilation quantity;
the fermentation medium comprises the following components:
60g/L glucose, 120g/L corn starch paste, 10g/L yeast extract, 10g/L oleic acid, 3g/L calcium carbonate, 1g/L magnesium sulfate heptahydrate, 1g/L dipotassium phosphate and 20mg/L, VB fulvic acid120mg/L，pH 7.0-7.2。
The preparation method of the fermentation medium comprises the following steps:
sequentially adding the raw materials into water, stirring, and adjusting pH.
The preparation method of the corn starch paste comprises the following steps:
adding water into corn starch to adjust the mass concentration of the starch milk to 50%, adjusting the pH to 5.5 and the temperature to 50 ℃ to prepare high-concentration starch milk; adding medium-temperature a-amylase (10000U/g) into high-concentration starch milk, wherein the addition amount of the a-amylase is 10U per gram of starch, heating to 60 ℃, performing enzymolysis for 50min under the stirring condition of 100rpm, heating to 100 ℃, inactivating the enzyme for 5min, and naturally cooling to room temperature to obtain the alpha-amylase.
Example 3
The influence of fermentation time and raw material addition in the fermentation medium on gum production.
Firstly, detecting the fermentation period of the strain, setting different fermentation times of 36,42,48,54,60,66,72 and 78 (h) respectively by taking the fermentation conditions of comparative example 1 as reference; as shown in figure 1, the fermentation time is close to the peak value after 66 hours, the time is increased continuously, the small increase is still realized, but the amplitude is small and is only 0.5%, the gum yield is not improved when the fermentation time reaches 78 hours, and the fermentation needs to be stopped at 72 hours at the moment to avoid generating more byproducts and causing difficulty in separation and extraction.
By comparison, the yields of the comparative example 1 and the comparative example 2 are relatively close to each other, namely 31.67g/L and 31.35g/L respectively, the yield of the comparative example 1 is slightly higher, but the fermentation liquor of the comparative example 2 cannot detect starch residues, and the yield of the comparative example 1 is 0.19g/L, so that the difficulty of subsequently separating and extracting xanthan gum is increased, and the improvement of the product quality is not facilitated.
Verifying the influence of the addition of eugenol in a fermentation medium on the gum yield, setting the concentrations to be 0, 0.1, 0.25, 0.5,1.0,1.5, 2.0,2.5,3.0,3.5 and 4.0 (mg/L), as shown in figure 2, when the concentration of eugenol is 0.1mg/L, the xanthan gum yield is hardly influenced, when the concentration of eugenol is continuously increased to 0.25mg/L, the xanthan gum yield is obviously improved, when the concentration of eugenol is not added, the xanthan gum yield is improved by 3.5 percent, when the concentration of eugenol is increased to 1.0mg/L, the xanthan gum yield is close to the highest value and is improved by 6.3 percent when the concentration of eugenol is not added, when the concentration of eugenol is increased to 2.0mg/L, the xanthan gum yield is slightly reduced, then the gum yield is obviously reduced along with the increase of the concentration, possibly because eugenol is an antibacterial agent and can inhibit the bacterial strain when the concentration exceeds a certain concentration and influence the normal growth activity of the xanthomonas bacterium, thereby leading the glue yield to slide down; in conclusion, it is more appropriate to select the concentration of eugenol to be lower than 2mg/L, and the optimal concentration range is 1-1.5 mg/L.
The foregoing list is only illustrative of the preferred embodiments of the present invention. It is obvious that the invention is not limited to the above embodiments, but that many variations are possible. All modifications which can be derived or suggested by a person skilled in the art from the disclosure of the present invention are to be considered within the scope of the invention.
Claims (10)
1. The fermentation process of the low-starch-residue xanthan gum product is characterized in that the fermentation medium used in the fermentation process comprises the following components: glucose, corn starch paste, yeast extract, oleic acid, calcium carbonate, magnesium sulfate heptahydrate, dipotassium hydrogen phosphate, fulvic acid and VB1And eugenol.
2. The fermentation process of claim 1, wherein the fermentation medium comprises:
60g/L glucose, 120g/L corn starch paste, 10g/L yeast extract, 10g/L oleic acid, 3g/L calcium carbonate, 1g/L magnesium sulfate heptahydrate, 1g/L dipotassium phosphate and 20mg/L, VB fulvic acid120mg/L, 1mg/L of eugenol and pH 7.0-7.2.
3. The fermentation process of claim 1, wherein the fermentation medium comprises:
60g/L glucose, 100g/L corn starch paste, 10g/L yeast extract, 10g/L oleic acid, 3g/L calcium carbonate, 1g/L magnesium sulfate heptahydrate, 1g/L dipotassium phosphate and 20mg/L, VB fulvic acid120mg/L, 1.5mg/L eugenol and pH 7.0-7.2.
4. The fermentation process of claims 1-3, wherein the fermentation medium is prepared by:
sequentially adding the raw materials into water, stirring, and adjusting pH.
5. The fermentation process of claim 4, wherein the corn starch paste is produced by enzymatic hydrolysis with an amylase.
6. The fermentation process of claim 5, wherein the corn starch paste is prepared by: adding water into corn starch to adjust the mass concentration of the starch milk to 50%, adjusting the pH to 5.5 and the temperature to 50 ℃ to prepare high-concentration starch milk; adding alpha-amylase into high-concentration starch milk, heating to 60 ℃, performing enzymolysis for 30-60min under the stirring condition of 100rpm, heating to 100 ℃, inactivating enzyme, and naturally cooling to room temperature to obtain the alpha-amylase.
7. The fermentation process of claim 6, wherein the a-amylase is added in an amount of 10U per gram of corn starch.
8. The fermentation process of claim 6, wherein the enzymolysis time is 40-50 min.
9. The fermentation process of claim 6, wherein the enzyme deactivation time is 5 min.
10. The fermentation process according to claims 1-9, wherein the fermentation process comprises in particular: inoculating the seed liquid of the xanthomonas into a fermentation tank filled with a fermentation culture medium for fermentation culture at the fermentation temperature of 30 ℃ for 66-72h to obtain fermentation liquid.
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