CN114277075A - Preparation method of peptone special for producing xanthan gum - Google Patents
Preparation method of peptone special for producing xanthan gum Download PDFInfo
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- CN114277075A CN114277075A CN202111491191.2A CN202111491191A CN114277075A CN 114277075 A CN114277075 A CN 114277075A CN 202111491191 A CN202111491191 A CN 202111491191A CN 114277075 A CN114277075 A CN 114277075A
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Abstract
The invention discloses a preparation method of peptone specially used for xanthan gum fermentation production, which comprises the following steps: mixing fresh fishbone with water, and steaming at high temperature and high pressure to obtain bone soup; pouring the bone soup into an enzymolysis tank, adding a compound enzyme, and stirring for enzymolysis; adding an acidity regulator after enzymolysis is finished, and regulating the pH value in the enzymolysis tank to subacidity to generate suspended matters and precipitates; and filtering clear liquid between the suspended matters and the precipitates, adding activated carbon powder, and performing centrifugal filtration and spray drying to obtain powder, thereby obtaining the peptone special for producing the xanthan gum. The invention prepares peptone by fresh fish bones and changes an enzymolysis process, so that the prepared peptone has higher content of amino acid nitrogen and amino acid composition suitable for xanthan gum strain fermentation, and the xanthan gum fermentation yield and quality are improved.
Description
Technical Field
The invention relates to the technical field of peptone, and particularly relates to a preparation method of peptone special for producing xanthan gum.
Background
Xanthan gum, also known as xanthan gum, is a microbial exopolysaccharide with a wide range of effects produced by Xanthomonas campestris through fermentation engineering with carbohydrates as main raw materials (such as corn starch). The xanthan gum has unique rheological property, good water solubility, stability to heat and acid and alkali, and good compatibility with various salts, can be widely applied to more than 20 industries such as food, petroleum, medicine and the like as a thickening agent, a suspending agent, an emulsifying agent and a stabilizing agent, is a microbial polysaccharide which has the largest production scale and extremely wide application in the world at present, is indispensable as a fermentation nitrogen source in the preparation process of the xanthan gum at present, is mainly prepared by normal enzymolysis through a plant protein source, an animal protein source or a microbial protein source, focuses on total nitrogen, and does not perform targeted development of an enzymolysis process aiming at the favor of xanthan gum fermentation strains on different amino acids, so that the xanthan gum fermentation yield and the product quality have a larger promotion space.
Disclosure of Invention
The invention aims to provide a preparation method of peptone special for producing xanthan gum.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows: a preparation method of peptone special for producing xanthan gum comprises the following steps:
s1, mixing fresh fishbone and water according to the weight ratio of 1: 4, mixing, and cooking at high temperature and high pressure to form bone soup;
s2, pouring the bone soup into an enzymolysis tank, and adding 0.1-0.3% of trypsin, 0.1-0.3% of aminopeptidase and 0.1-0.3% of neutral protease for stirring and enzymolysis;
s3, adding an acidic regulator after enzymolysis is finished, regulating the pH value in the enzymolysis tank to subacidity, and performing temperature rising and cooling operations to generate suspended matters and precipitates;
s4, filtering clear liquid between the suspended matters and the sediments by 400-mesh filter cloth, intercepting filtered liquid by a 1.5KDa ultrafiltration membrane, concentrating the intercepted liquid in vacuum until the solid content is 50%, adding activated carbon powder, and drying the activated carbon powder into powder by a centrifugal filter and a spray dryer to obtain the peptone special for producing xanthan gum.
Preferably, the pressure in the step S1 is 2-3 MPa, and the temperature is 100-120 ℃.
Preferably, in step S2, the pH of the bone soup is adjusted to 7-7.5 and the temperature is 50 ℃ after the bone soup is poured into the enzymolysis tank.
Preferably, the stirring in step S2 is divided into multiple times of stirring, wherein the first stirring is stopped for 5 minutes for 10 minutes, the second stirring is stopped for 5 minutes for 10 minutes, the third stirring is stopped for 10 minutes, the fourth stirring is stopped for 10 minutes, the fifth stirring is stopped for 10 minutes and 25 minutes for 10 minutes, and the sixth stirring is stopped for 25 minutes for 10 minutes.
Preferably, the specific steps of step S3 are: the acid regulator is acetic acid, the pH value of the solution in the enzymolysis tank is regulated to 5.1-5.3, the temperature of the dissolution tank is slowly reduced after being raised to 90 ℃, the temperature is reduced to 45-55 ℃ for 2-3 hours, and stirring cannot be started in the process of reducing the temperature.
Preferably, the total nitrogen content of the prepared peptone is 13.5-15.5%, wherein the amino acid nitrogen content is 2.5-4%.
A method for preparing xanthan gum of the peptone comprises the following steps: adding the peptone and the culture fermentation solution into a fermentation tank, inoculating the xanthomonas campestris, and fermenting at the pH of 7.0 and the temperature of 28-35 ℃; extracting the fermentation liquor by a salt precipitation method to obtain a xanthan gum precipitate; adding heat-resistant additive into the extracted xanthan gum precipitate, and then performing solid-liquid separation, dehydration and drying to obtain the xanthan gum.
Preferably, the culture fermentation solution is a mixed solution of 50-60 g/L of corn starch, (K2HPO4.3H2O) 3-5 g/L, (MgSO4.7H2O) 0.1-0.4 g/L, (NH4)2SO 41-5 g/L and 2-4 g/L of citric acid, wherein the dilution rate n = 5.
Preferably, the salt precipitation method employs a 25% potassium chloride solution.
Compared with the prior art, the invention has the advantages that:
the method has the advantages that the production cost can be reduced by preparing the peptone from the fresh fish bones, the enzymolysis efficiency can be increased and the pigment content can be reduced by changing the stirring mode, changing the proportion of enzyme and increasing the active carbon powder in the preparation process, the yield is greatly improved, and the peptone has higher content of amino acid nitrogen and amino acid composition suitable for fermentation of xanthan gum strains, is suitable for preparing the xanthan gum and improves the gum yield of the xanthan gum by changing the process.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a graph of molecular weight measurements of examples of the present invention and comparative examples;
FIG. 2 is a graph comparing the amount of gum formed in examples of the present invention and comparative examples.
Detailed Description
The preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings so that the advantages and features of the present invention can be more easily understood by those skilled in the art, and the scope of the present invention will be more clearly and clearly defined.
Example one
A preparation method of peptone special for producing xanthan gum comprises the following steps:
fresh fishbone and water are mixed according to the weight ratio of 1: 4 mixing, and steaming under the pressure of 2MPa and at the temperature of 105 ℃ to obtain bone soup;
pouring the bone soup into an enzymolysis tank, adjusting the pH value to 7, adjusting the temperature to 50 ℃, adding 0.1% of trypsin, 0.1% of aminopeptidase and 0.3% of neutral protease, stirring and carrying out enzymolysis, wherein the stirring is divided into a plurality of times of stirring, the first stirring is firstly carried out for 10 minutes and is stopped for 5 minutes, the second stirring is carried out for 10 minutes, the third stirring is carried out for 10 minutes, the fourth stirring is carried out for 10 minutes, the fifth stirring is carried out for 10 minutes and is stopped for 25 minutes, and the sixth stirring is carried out for 10 minutes and is stopped for 25 minutes;
after enzymolysis, adding acetic acid, adjusting the pH value of the solution in the enzymolysis tank to 5.1, slowly cooling the solution in the dissolving tank after the temperature of the dissolving tank is raised to 90 ℃, reducing the temperature to 45 ℃ for 2 hours, and not stirring the solution in the cooling process to generate suspended matters and precipitates;
and filtering clear liquid between the suspended matters and the precipitates by 400-mesh filter cloth, intercepting filtered liquid by a 1.5KDa ultrafiltration membrane, concentrating the intercepted liquid in vacuum until the solid content is 50%, adding activated carbon powder, performing centrifugal filtration and drying by a spray dryer to obtain the peptone special for producing the xanthan gum.
The molecular weight distribution of the peptone is 200-1200Da, and the total nitrogen content of the peptone is 15.4 percent, wherein the amino acid nitrogen content is 3.8 percent.
Example two
A preparation method of peptone special for producing xanthan gum comprises the following steps:
fresh fishbone and water are mixed according to the weight ratio of 1: 4 mixing, and steaming under the pressure of 2.5MPa and at the temperature of 110 ℃ to form bone soup;
pouring the bone soup into an enzymolysis tank, adjusting the pH value to 7, adjusting the temperature to 50 ℃, adding 0.1% of trypsin, 0.3% of aminopeptidase and 0.1% of neutral protease, stirring and carrying out enzymolysis, wherein the stirring is divided into a plurality of times of stirring, the first stirring is carried out for 10 minutes and stopped for 5 minutes, the second stirring is carried out for 10 minutes and stopped for 5 minutes, the third stirring is carried out for 10 minutes, the fourth stirring is carried out for 10 minutes, the fifth stirring is carried out for 10 minutes and stopped for 25 minutes, and the sixth stirring is carried out for 10 minutes and stopped for 25 minutes;
after enzymolysis, adding acetic acid, adjusting the pH value of the solution in the enzymolysis tank to 5.2, slowly cooling the solution in the dissolving tank after the temperature of the dissolving tank is raised to 90 ℃, reducing the temperature to 50 ℃ within 2.5 hours, and not stirring the solution in the cooling process to generate suspended matters and precipitates;
and filtering clear liquid between the suspended matters and the precipitates by 400-mesh filter cloth, intercepting filtered liquid by a 1.5KDa ultrafiltration membrane, concentrating the intercepted liquid in vacuum until the solid content is 50%, adding activated carbon powder, performing centrifugal filtration and drying by a spray dryer to obtain the peptone special for producing the xanthan gum.
The molecular weight distribution of the peptone is 100-1100Da, and the total nitrogen content of the peptone is 15.5 percent, wherein the amino acid nitrogen content is 4 percent.
EXAMPLE III
A preparation method of peptone special for producing xanthan gum comprises the following steps:
fresh fishbone and water are mixed according to the weight ratio of 1: 4 mixing, and steaming under the conditions of 3MPa pressure and 120 ℃ to obtain bone soup;
pouring the bone soup into an enzymolysis tank, adjusting the pH value to 7.5, adjusting the temperature to 50 ℃, adding 0.3% of trypsin, 0.1% of aminopeptidase and 0.1% of neutral protease, stirring for enzymolysis, wherein the stirring is divided into a plurality of times of stirring, the first stirring is firstly stopped for 5 minutes in 10 minutes, the second stirring is stopped for 5 minutes in 10 minutes, the third stirring is stopped for 10 minutes, the fourth stirring is stopped for 10 minutes, the fifth stirring is stopped for 25 minutes in 10 minutes, and the sixth stirring is stopped for 25 minutes in 10 minutes;
after enzymolysis, adding acetic acid, adjusting the pH value of the solution in the enzymolysis tank to 5.3, slowly cooling the solution in the dissolving tank after the temperature of the dissolving tank is raised to 90 ℃, reducing the temperature to 55 ℃ for 3 hours, and not stirring the solution in the cooling process to generate suspended matters and precipitates;
and filtering clear liquid between the suspended matters and the precipitates by 400-mesh filter cloth, intercepting filtered liquid by a 1.5KDa ultrafiltration membrane, concentrating the intercepted liquid in vacuum until the solid content is 50%, adding activated carbon powder, performing centrifugal filtration and drying by a spray dryer to obtain the peptone special for producing the xanthan gum.
The molecular weight distribution of the peptone is 150-1200Da, and the total nitrogen content of the peptone is 15.6 percent, wherein the amino acid nitrogen content is 3.5 percent.
Comparative example 1
The difference between the first comparative example and the second example is that the first comparative example is different in stirring mode, wherein the first stirring is performed for 10 minutes after 5 minutes, the second stirring is performed for 10 minutes after 5 minutes, the third stirring is performed for 15 minutes after 5 minutes, the fourth stirring is performed for 15 minutes after 5 minutes, the fifth stirring is performed for 5 minutes and 30 minutes after 5 minutes, and the sixth stirring is performed for 5 minutes and 30 minutes after 5 minutes;
the molecular weight distribution of the peptone is 200-1300Da, and the total nitrogen content of the peptone is 14.5 percent, wherein the amino acid nitrogen content is 3.6 percent.
Comparative example No. two
The difference between the comparative example II and the example II is that 0.5% trypsin is added as a single enzyme in the comparative example II, the molecular weight distribution of the peptone is 300-1500Da, and the total nitrogen content of the peptone prepared is 14.8%, wherein the amino acid nitrogen content is 3%.
Comparative example No. three
The difference between the third comparative example and the second example is that 0.5% aminopeptidase was added as a single enzyme in the second comparative example, the molecular weight distribution of the peptone was 200-1100Da, and the total nitrogen content of the peptone produced was 15% with the amino acid nitrogen content of 3.2%.
Comparative example No. four
The difference between the comparative example four and the example two is that the single enzyme 0.5% neutral protease is added in the comparative example two, the molecular weight distribution of the peptone is 200-1200Da, and the total nitrogen content of the peptone is 15.1%, wherein the amino acid nitrogen content is 3.1%.
The peptone powders prepared in examples one to three and comparative examples one to four all have light yellow color and good physical properties.
The molecular weight distributions of the finished products of the peptone powders of examples one to three and comparative examples one to four were measured by the SDS-PAGE method. The results show that the peptones of the above groups all have molecular weight distribution less than or equal to 1500Da, and the difference of the molecular weight distribution is small, so that the quality stability of the product can be ensured, specifically as shown in FIG. 1, the total nitrogen content and the amino acid nitrogen content of the peptones in the examples are higher than those of the comparative examples, while the fermentation strain of xanthan gum has better growth effect in high amino acid nitrogen, high gum yield, and no precipitation appears when the aqueous solution of the peptone powder is dropped into saturated sodium sulfate, which indicates that the peptone product per se meets the requirements of standard peptones.
The peptones of examples one to three and comparative examples one and four were subjected to the following procedure for the preparation of xanthan gum: adding the prepared peptone and a culture fermentation solution into a fermentation tank, wherein the culture fermentation solution is a mixed solution of 60g/L of corn starch, (K2HPO4.3H2O) 4g/L, (MgSO4.7H2O) 0.3 g/L, (NH4)2SO 44 g/L and 34g/L of citric acid, and the dilution rate n = 5; inoculating M-137 Xanthomonas campestris, and fermenting at 30 deg.C and pH 7.0; extracting the fermentation liquor by using 25% potassium chloride solution to obtain xanthan gum precipitate; adding heat-resistant additive into the extracted xanthan gum precipitate, and then performing solid-liquid separation, dehydration and drying to obtain xanthan gum, which is shown in figure 2.
As can be seen from fig. 2, the xanthan gum yield is the highest when the xanthan gum is cultured in the second example, which has the best culture performance and the culture time is shorter, and the first example and the third example are followed; in the first comparative example, the stirring time is changed, ammonia nitrogen and total nitrogen are reduced, the ammonia nitrogen is higher than that in the third example, but the gum yield is lower than that in the third example, and the reason is probably caused by difference in amino acid composition; compared with the second to the fourth, the single enzyme is added, the hydrolysis effect is not good as that of the complex enzyme, and the glue yield is not high as that of the embodiment.
Although the embodiments of the present invention have been described with reference to the accompanying drawings, various changes or modifications may be made by the patentees within the scope of the appended claims, and within the scope of the invention, as long as they do not exceed the scope of the invention described in the claims.
Claims (9)
1. A preparation method of peptone special for producing xanthan gum is characterized by comprising the following steps:
s1, mixing fresh fishbone and water according to the weight ratio of 1: 4, mixing, and cooking at high temperature and high pressure to form bone soup;
s2, pouring the bone soup into an enzymolysis tank, and adding 0.1-0.3% of trypsin, 0.1-0.3% of aminopeptidase and 0.1-0.3% of neutral protease for stirring and enzymolysis;
s3, adding an acidic regulator after enzymolysis is finished, regulating the pH value in the enzymolysis tank to subacidity, and performing temperature rising and cooling operations to generate suspended matters and precipitates;
s4, filtering clear liquid between the suspended matters and the sediments by 400-mesh filter cloth, intercepting filtered liquid by a 1.5KDa ultrafiltration membrane, concentrating the intercepted liquid in vacuum until the solid content is 50%, adding activated carbon powder, and drying the activated carbon powder into powder by a centrifugal filter and a spray dryer to obtain the peptone special for producing xanthan gum.
2. The preparation method of peptone specially used for xanthan gum according to claim 1, characterized in that: in the step S1, the pressure is 2-3 MPa, and the temperature is 100-120 ℃.
3. The preparation method of peptone specially used for xanthan gum according to claim 1, characterized in that: in step S2, the pH value of the bone soup is adjusted to 7-7.5 and the temperature is 50 ℃ after the bone soup is poured into the enzymolysis tank.
4. The preparation method of peptone specially used for xanthan gum according to claim 1, characterized in that: in step S2, the stirring is divided into multiple times of stirring, wherein the first stirring is stopped for 5 minutes within 10 minutes, the second stirring is stopped for 5 minutes within 10 minutes, the third stirring is stopped for 10 minutes within 10 minutes, the fourth stirring is stopped for 10 minutes within 10 minutes, the fifth stirring is stopped for 10 minutes within 25 minutes, and the sixth stirring is stopped for 10 minutes within 25 minutes.
5. The preparation method of peptone specially used for xanthan gum according to claim 1, characterized in that: the specific steps of step S3 are: the acid regulator is acetic acid, the pH value of the solution in the enzymolysis tank is regulated to 5.1-5.3, the temperature of the dissolution tank is slowly reduced after being raised to 90 ℃, the temperature is reduced to 45-55 ℃ for 2-3 hours, and stirring cannot be started in the process of reducing the temperature.
6. The preparation method of peptone specially used for xanthan gum according to claim 1, characterized in that: the total nitrogen content of the prepared peptone is 13.5-15.5%, wherein the amino acid nitrogen content is 2.5-4%.
7. A process for the preparation of xanthan gum using peptones according to claims 1 to 6, characterized by the following steps: adding the peptone and the culture fermentation solution into a fermentation tank, inoculating the xanthomonas campestris, and fermenting at the pH of 7.0 and the temperature of 28-35 ℃; extracting the fermentation liquor by a salt precipitation method to obtain a xanthan gum precipitate; adding heat-resistant additive into the extracted xanthan gum precipitate, and then performing solid-liquid separation, dehydration and drying to obtain the xanthan gum.
8. The method of preparing xanthan gum according to claim 7, wherein: the culture fermentation solution is 50-60 g/L (K) of corn starch2HPO4•3H2O)3~5g/L、(MgSO4•7H2O)0.1~0.4 g/L、(NH4)2SO4 1-5 g/L and 2-4 g/L citric acid, wherein the dilution rate n = 5.
9. A process for the preparation of xanthan gum according to claim 1, wherein: the salt precipitation method uses 25% potassium chloride solution.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117587083A (en) * | 2023-12-19 | 2024-02-23 | 内蒙古工业大学 | Fermentation method of xanthan gum |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117587083A (en) * | 2023-12-19 | 2024-02-23 | 内蒙古工业大学 | Fermentation method of xanthan gum |
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