CN117229958A - Xanthomonas campestris and application thereof in preparing low-viscosity xanthan gum - Google Patents
Xanthomonas campestris and application thereof in preparing low-viscosity xanthan gum Download PDFInfo
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- xanthan gum
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- 239000000230 xanthan gum Substances 0.000 title claims abstract description 71
- 229920001285 xanthan gum Polymers 0.000 title claims abstract description 71
- 229940082509 xanthan gum Drugs 0.000 title claims abstract description 71
- 235000010493 xanthan gum Nutrition 0.000 title claims abstract description 71
- 241000589636 Xanthomonas campestris Species 0.000 title claims abstract description 45
- 238000000855 fermentation Methods 0.000 claims abstract description 74
- 230000004151 fermentation Effects 0.000 claims abstract description 73
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 36
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 24
- 238000000034 method Methods 0.000 claims abstract description 24
- 229910052757 nitrogen Inorganic materials 0.000 claims abstract description 18
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 16
- 238000002360 preparation method Methods 0.000 claims abstract description 13
- 238000004321 preservation Methods 0.000 claims abstract description 11
- 238000005273 aeration Methods 0.000 claims abstract description 10
- 230000000813 microbial effect Effects 0.000 claims abstract description 10
- 238000012258 culturing Methods 0.000 claims abstract description 8
- 235000015097 nutrients Nutrition 0.000 claims abstract description 8
- 238000006243 chemical reaction Methods 0.000 claims description 58
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 40
- 102000016943 Muramidase Human genes 0.000 claims description 39
- 108010014251 Muramidase Proteins 0.000 claims description 39
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 claims description 39
- 239000004325 lysozyme Substances 0.000 claims description 39
- 229960000274 lysozyme Drugs 0.000 claims description 39
- 235000010335 lysozyme Nutrition 0.000 claims description 39
- 108091005658 Basic proteases Proteins 0.000 claims description 31
- 239000002253 acid Substances 0.000 claims description 30
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 28
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 28
- 238000000108 ultra-filtration Methods 0.000 claims description 25
- 229940088598 enzyme Drugs 0.000 claims description 24
- 102000004190 Enzymes Human genes 0.000 claims description 23
- 108090000790 Enzymes Proteins 0.000 claims description 23
- 230000001105 regulatory effect Effects 0.000 claims description 22
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 21
- 239000000706 filtrate Substances 0.000 claims description 21
- 238000001914 filtration Methods 0.000 claims description 18
- 239000007788 liquid Substances 0.000 claims description 17
- 150000003839 salts Chemical class 0.000 claims description 17
- 239000012295 chemical reaction liquid Substances 0.000 claims description 15
- 230000000694 effects Effects 0.000 claims description 15
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 14
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims description 14
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 14
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical class O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 14
- 238000010438 heat treatment Methods 0.000 claims description 14
- 229910017604 nitric acid Inorganic materials 0.000 claims description 14
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims description 14
- 229920002472 Starch Polymers 0.000 claims description 13
- 239000007787 solid Substances 0.000 claims description 13
- 235000019698 starch Nutrition 0.000 claims description 13
- 239000008107 starch Substances 0.000 claims description 13
- 238000010008 shearing Methods 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 229930006000 Sucrose Natural products 0.000 claims description 9
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 9
- 239000005720 sucrose Substances 0.000 claims description 9
- 230000000415 inactivating effect Effects 0.000 claims description 8
- 239000002994 raw material Substances 0.000 claims description 8
- 102000004157 Hydrolases Human genes 0.000 claims description 7
- 108090000604 Hydrolases Proteins 0.000 claims description 7
- 240000008042 Zea mays Species 0.000 claims description 7
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 7
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 7
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 7
- 229940041514 candida albicans extract Drugs 0.000 claims description 7
- 210000002421 cell wall Anatomy 0.000 claims description 7
- 238000001816 cooling Methods 0.000 claims description 7
- 235000005822 corn Nutrition 0.000 claims description 7
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 7
- 239000002054 inoculum Substances 0.000 claims description 7
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 7
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 7
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 7
- 239000004317 sodium nitrate Substances 0.000 claims description 7
- 235000010344 sodium nitrate Nutrition 0.000 claims description 7
- 239000008223 sterile water Substances 0.000 claims description 7
- 238000009423 ventilation Methods 0.000 claims description 7
- 239000012138 yeast extract Substances 0.000 claims description 7
- 239000002518 antifoaming agent Substances 0.000 claims description 6
- 238000000605 extraction Methods 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 3
- 238000001694 spray drying Methods 0.000 claims description 3
- 239000012530 fluid Substances 0.000 claims description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 2
- 230000007071 enzymatic hydrolysis Effects 0.000 claims 2
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims 2
- 230000002255 enzymatic effect Effects 0.000 claims 1
- 150000003384 small molecules Chemical class 0.000 claims 1
- 235000013361 beverage Nutrition 0.000 abstract description 8
- 150000004676 glycans Chemical class 0.000 abstract description 3
- 229920001282 polysaccharide Polymers 0.000 abstract description 3
- 239000005017 polysaccharide Substances 0.000 abstract description 3
- 239000002609 medium Substances 0.000 description 22
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 18
- 239000001103 potassium chloride Substances 0.000 description 9
- 235000011164 potassium chloride Nutrition 0.000 description 9
- 235000013373 food additive Nutrition 0.000 description 5
- 239000002778 food additive Substances 0.000 description 5
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 5
- 238000005259 measurement Methods 0.000 description 4
- 239000000376 reactant Substances 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 238000000746 purification Methods 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 239000003513 alkali Substances 0.000 description 2
- 230000000593 degrading effect Effects 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000009629 microbiological culture Methods 0.000 description 2
- 125000001477 organic nitrogen group Chemical group 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 108010077805 Bacterial Proteins Proteins 0.000 description 1
- 241000589634 Xanthomonas Species 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 238000005266 casting Methods 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000013530 defoamer Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000013325 dietary fiber Nutrition 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 239000000416 hydrocolloid Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000001965 increasing effect Effects 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000004753 textile Substances 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Abstract
The application belongs to the preparation of microbial polysaccharide, and particularly relates to Xanthomonas campestris and application thereof in preparing low-viscosity xanthan gum. The method comprises inoculating Xanthomonas campestris with Latin name Xanthomonas campestris and preservation number of CGMCC No.17845 into fermentation medium containing carbon source, nitrogen source and nutrient element, culturing under aeration to obtain fermentation broth, and collecting low viscosity xanthan gum from the fermentation broth. The application solves the technical problems that the xanthan gum prepared by the prior art is relatively high in viscosity and not suitable for preparing the refreshing beverage, and the like, and has the advantages that the prepared xanthan gum meets the requirements of the prior national standard, the viscosity value is obviously lower than that of the prior product, and the xanthan gum is suitable for preparing the refreshing beverage, and the like.
Description
Technical Field
The application belongs to the preparation of microbial polysaccharide, and particularly relates to Xanthomonas campestris and application thereof in preparing low-viscosity xanthan gum.
Background
According to the description of "Xanthan Gum" food additive (GB 1886.41-2015), xanthan Gum is a microbial polysaccharide obtained by taking Xanthomonas glabra as a producer, taking starch sugar as a main raw material, carrying out specific biological fermentation, purifying, drying and crushing.
The xanthan gum has excellent rheological property due to unique molecular structure, has good viscosity increasing property, acid and alkali resistance, pseudoplasticity and high temperature resistance, has emulsifying and suspending properties, is used as a stabilizer, a thickener, an emulsifier and a suspending agent, and is widely applied to industries such as food, medicine, petroleum, textile, casting and the like. With the improvement of the living standard of people and the development of beverage industry, the requirements of consumers on beverages are also higher and higher, and the consumers are more and more inclined to be refreshing, healthy and delicious. Xanthan gum is a widely used hydrocolloid that is soluble in cold and hot water to form a high viscosity solution with high suspension stability. However, the high viscosity of xanthan gum also seriously affects the taste of the beverage, which makes the beverage burned and has poor flavor release.
The xanthan gum has good resistance to acid, alkali, heat, enzyme and the like due to the special double-helix structure, and is not easy to degrade. According to the requirements of food additive xanthan gum (GB 1886.41-2015), the viscosity of 1% potassium chloride solution of xanthan gum is more than or equal to 600cp, the viscosity of 1% potassium chloride solution of xanthan gum in the market at present is mainly high-viscosity xanthan gum, the viscosity of 1% potassium chloride solution is more than 1400cp, the research on the production of low-viscosity xanthan gum is less, ultrasonic waves with power of 350W, frequency of 20Hz and the like are used for 1% xanthan gum (2% H is added) 2 O 2 ) The low-viscosity dietary fiber is prepared by degrading the xanthan gum by ultrasonic waves after the action for 3 hours, so that the viscosity of the 1% potassium chloride solution of the xanthan gum can reach 600cp. However, due to the fact that the ultrasonic attenuation and the power are not matched with the equipment, blank areas and the like are easy to generate, and the method is not suitable for large-scale production.
Disclosure of Invention
One of the purposes of the application is to provide a Xanthomonas campestris strain, the Latin name of which is Xanthomonas campestris and the preservation number of which is CGMCC No.17845. The strain can use sucrose, starch sugar and other carbon sources, and organic nitrogen and inorganic nitrogen as nitrogen sources, and has the advantages of strong activity, small grafting amount, high yield, 42g/L of yield, low viscosity of the produced xanthan gum, and 900-1200 cp of untreated 1% potassium chloride solution.
The strain in the application is submitted to China general microbiological culture Collection center for collection of China Committee of China number 1 and number 3 of North Chen West Lu 1 in the Korean area of Beijing city for 22 days in 2019, and the collection unit is called CGMCC for short and has the collection number: CGMCC No.17845.
The applicant obtains the strain in the application after multiple mutagenesis, separation and purification based on Xanthomonas campestris Xanthomonas campestris CGMCC No.10122, the strain is gram negative bacteria and short rod-shaped, can form capsule, and bacterial colony is light yellow, round bump, smooth surface and viscous liquid. The xanthan gum prepared by adopting the strain has lower viscosity.
The second purpose of the application is to provide the application of Xanthomonas campestris in preparing the xanthan gum with low viscosity.
The specific technical conception of the application is as follows:
the application of Xanthomonas campestris in preparing the low-viscosity xanthan gum comprises the following steps:
A. inoculating Latin name Xanthomonas campestris and accession number CGMCC No.17845 Xanthomonas campestris into fermentation medium containing carbon source, nitrogen source and nutrient element, and culturing under aeration to obtain fermentation broth;
B. and C, collecting the low-viscosity xanthan gum from the fermentation liquor prepared in the step A.
The main function of high ventilation fermentation is to facilitate the full utilization of culture medium and further improve the purity of xanthan gum, and the preferred technical implementation means is that the ventilation in the ventilation culture in the step A is 500m 3 /h～4200m 3 /h。
The more preferable technical implementation means is that the ventilation quantity in the step A is as follows:
0 to 10 hoursWhen (1): air volume 500m 3 /h～1200m 3 /h；
11-20 hours: air volume 1200m 3 /h～2000m 3 /h；
21-30 hours: air volume 2000m 3 /h～2800m 3 /h；
31-40 hours: air volume 2800m 3 /h～3600m 3 /h；
41-50 hours: air volume 3600m 3 /h～4200m 3 /h；
51 hours to tank discharge: air volume 4200m 3 /h～3800m 3 /h。
In order to facilitate the purification of the xanthan gum, the preferred technical implementation means is that the fermentation medium in the step A takes sucrose and/or starch sugar as a carbon source and inorganic and organic micromolecular nitrogen sources as nitrogen sources.
The more preferable technical implementation mode is that the fermentation medium in the step A consists of the following raw materials in percentage by mass:
3% -5% of starch sugar and/or sucrose; sodium nitrate 0.15-0.3%; corn steep liquor 0.2% -0.3%; yeast extract 0.03-0.06%; 0.1 to 0.2 percent of monopotassium phosphate; 0.1 to 0.2 percent of dipotassium hydrogen phosphate; 0.015 to 0.025 percent of magnesium sulfate; 0.03 to 0.05 percent of defoaming agent; the balance being sterile water.
In order to optimize the fermentation process and reduce the fermentation seed investment, the preferred technical implementation means is that Latin name Xanthomonas campestris and the preservation number of CGMCC No.17845 are subjected to enlarged culture in the step A, and then the Xanthomonas campestris is subjected to the enlarged culture according to the seed liquid: fermentation medium = 5% -15% inoculum size seed fluid is inoculated into sterilized fermentation medium.
In order to degrade the molecular chain of xanthan gum, reduce the viscosity of xanthan gum, remove salt and other small organic molecules in solution and improve the concentration of xanthan gum, the preferred technical implementation means is that the step B comprises the following process steps:
b1, enzymolysis reaction: regulating the pH of the prepared fermentation liquor, adding alkaline protease for enzymolysis reaction, regulating the pH after the reaction is finished, and adding lysozyme for enzymolysis reaction;
b2, inactivating the enzyme preparation: regulating the pH value of the enzymolysis liquid after the enzymolysis reaction in the step B to be 1.5-3.0, heating to 80-95 ℃ and preserving heat for 30-60 minutes;
b3, decoloring and filtering: cooling the reaction liquid prepared in the step C to 75-85 ℃, regulating the pH value to be 6.5-7.5, decoloring, adding a filter aid into the decolored reaction liquid, filtering, and collecting filtrate;
b4, shearing and ultrafiltering: and D, heating the filtrate prepared in the step D, homogenizing the heated reaction liquid under high pressure, and then performing ultrafiltration concentration until the solid content is 8-12%.
In order to hydrolyze the mycoprotein and other water-insoluble organic high molecular substances into water-soluble small molecular substances so as to expose the molecular chain of the xanthan gum and be easy to degrade, the preferred technical implementation mode is that the alkaline protease treatment condition in the step B1 is that 0.02-0.1% of alkaline protease is added according to the amount of fermentation liquor, and the reaction is carried out for 3-5 hours; the lysozyme treatment condition is that 0.01 to 0.05 percent of lysozyme is added according to the amount of the fermentation liquid, and the temperature is kept for 2 to 4 hours.
Further, the enzyme activity of the alkaline protease is 20 mu/mg, the enzymolysis reaction temperature of the alkaline protease is 50-60 ℃, and the pH=9.5-10.5; the lysozyme adopts microbial cell wall hydrolase-glycoside hydrolase, the enzyme activity is 2 mu/mg, the temperature of the enzymolysis reaction of the lysozyme is 30-40 ℃, and the pH value is 4.5-6.5.
The more preferable technical implementation means is that in the step B1, naOH solution or KOH solution with the mass percent concentration of 10-20% is used for adjusting the pH value in the enzymolysis reaction of alkaline protease; the pH is adjusted by 10-20% acid solution in the enzymolysis reaction of lysozyme, and the acid is one of hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid and nitric acid.
In order to realize the degradation of the molecular chain of the xanthan gum and reduce the viscosity of the xanthan gum while inactivating enzyme, the preferable technical implementation means is that 10% -20% of acid solution is used for adjusting the pH value to be 1.5-3.0 in the step B2, and the acid is hydrochloric acid, sulfuric acid or nitric acid.
In order to increase the filtering effect, the preferable technical implementation means is that 10-20% NaOH solution or KOH solution with mass percent concentration is used for adjusting the pH value to be 6.5-7.5 in the step B3; decolorizing with 1-3% active carbon for 30-40 min; adding filter aid accounting for 1-2% of the total volume, filtering, and collecting filtrate; the filter aid adopts a mixture of diatomite and pearl salt, and the diatomite: mass ratio of pearl salt=2 to 4:6 to 8.
In order to further reduce the viscosity of the xanthan gum, improve the concentration of the xanthan gum and remove salt and other small organic molecules in the solution, the preferable technical implementation means is that in the step B4, the filtrate is heated to 70-80 ℃ and homogenized under the condition of 30 Mpa; the ultrafiltration condition is that the molecular weight cut-off is 50000 daltons, the ultrafiltration pressure is 0.2-0.3 MPa, and the concentration is carried out until the solid content is 8-12%.
The method also comprises a step B5, wherein the process conditions of the step are as follows:
and B5, spray drying or alcohol extraction, drying and crushing the reaction liquid subjected to ultrafiltration concentration in the step B4 to prepare the solid low-viscosity xanthan gum.
To verify the viscosity of the xanthan gum prepared by the method of the application, the applicant carried out the following tests:
1. determination of viscosity
The preparation was carried out by the method described in "Xanthan Gum as food additive" (GB 1886.41-2015).
1 determination of viscosity
1.1 instruments and apparatus
A Brookfield rotational viscometer or other equivalent performance viscometer.
1.2 measurement conditions
1.2.1 rotor model: rotor No. 3.
2.2.2 rotor speed: 60 revolutions per minute.
1.2.3 measurement temperature: 24-25 ℃.
1.3 analytical procedure
1.3.1 preparation of a solution containing 1% sample and 1% Potassium chloride
1.3.1.1 3g of the sample and potassium chloride (accurate to 0.01 g) were weighed separately with clean, dry weighing paper and mixed well.
1.3.1.2 measuring 300ml distilled water into a 400ml beaker.
1.3.1.3 placing the beaker containing water under a stirrer, starting the stirrer, slowly adding the mixed sample into the water between the stirring blade and the wall of the cup, starting timing, continuously stirring at 800 revolutions per minute for 2 hours, and keeping the temperature at 24-25 ℃.
1.3.1.4 the stirring is stopped, the cup is removed, and the solution is turned up and down with a stirring rod or the like.
1.3.2 determination
An appropriate amount of a solution containing 1% of the sample and 1% of potassium chloride was placed in a 100ml high-format beaker, and the measurement was performed under predetermined measurement conditions.
2. Determination of the shear Property values
2.1 measurement method
The viscosity values of rotor number 3 at 6 rpm and 60 rpm were determined at 1.3, respectively.
2.2 calculation of results
Shear performance value N, calculated as:
N＝η 1 /η 2
wherein:
η 1 -viscosity value at 6 revolutions per minute in centipoise (cP);
η 2 -viscosity value at 60 revolutions per minute in centipoise (cP).
The experimental results are as follows:
the low-viscosity xanthan gum produced by the application has the viscosity of 600-800 cP and the shearing performance value of more than 7.2, has a certain suspension function, greatly reduces the viscosity compared with the common xanthan gum, and is suitable for preparing fresh beverage according to the method described in food additive xanthan gum (GB 1886.41-2015).
The essential characteristics and remarkable technical progress of the application are as follows:
1. the application provides a Xanthomonas campestris strain, which can utilize carbon sources such as sucrose, starch sugar and the like, and organic nitrogen and inorganic nitrogen are utilized as nitrogen sources, so that the produced Xanthan gum not only meets the performance index requirement of food additive Xanthan Gum (GB 1886.41-2015) on Xanthan Gum, but also has lower viscosity and higher shearing performance value.
2. In the application, sucrose and/or starch sugar are used as carbon sources and inorganic and organic micromolecular nitrogen sources are used as nitrogen sources in a fermentation medium, so that the purification of xanthan gum is easy to realize; the high ventilation rate fermentation is adopted in the fermentation process, so that the full utilization of the culture medium is facilitated, and the purity of the xanthan gum is further improved.
3. In the application, in the post-fermentation extraction, diluted fermentation liquor is subjected to double-enzyme hydrolysis by alkaline protease and lysozyme, so that bacterial protein and other water-insoluble organic high molecular substances are hydrolyzed into water-soluble small molecular substances, and a xanthan gum molecular chain is exposed and is easy to degrade;
4. in the extraction after fermentation, the pH=1.5-3.0 and the temperature of 80-95 ℃ are controlled, so that the enzyme preparation is inactivated; and secondly, degrading a xanthan gum molecular chain to reduce the viscosity of the xanthan gum.
5. The application uses high temperature and high pressure to carry out the homogenization shearing treatment, which can further reduce the viscosity of the xanthan gum.
6. The application adopts ultrafiltration concentration, and effectively removes salt and other small organic molecules in the solution while improving the concentration of the xanthan gum.
7. The low-viscosity xanthan gum produced by the application is detected by the method recorded by the current national standard, the viscosity of the 1% potassium chloride solution is 600-800 cP, the shearing performance value is above 7.2, and the low-viscosity xanthan gum has a certain suspension function, and meanwhile, the viscosity is greatly reduced compared with that of the common xanthan gum, so that the low-viscosity xanthan gum is suitable for preparing fresh beverage.
The strain in the application is submitted to China general microbiological culture Collection center for collection of China Committee of China number 1 and number 3 of North Chen West Lu 1 in the Korean area of Beijing city for 22 days in 2019, and the collection unit is called CGMCC for short and has the collection number: CGMCC No.17845.
Detailed Description
The present application is further described below with reference to examples, but the present application is not limited thereto, and the claims of the present application should be construed as being limited thereto, and any equivalent means according to the specification may be substituted without departing from the scope of the present application.
Example 1
The application of Xanthomonas campestris in preparing the low-viscosity xanthan gum comprises the following steps:
A. inoculating Latin name Xanthomonas campestris and accession number CGMCC No.17845 Xanthomonas campestris into fermentation medium containing carbon source, nitrogen source and nutrient element, and culturing under aeration to obtain fermentation broth;
B. and C, collecting the low-viscosity xanthan gum from the fermentation liquor prepared in the step A.
The ventilation quantity in the step A is as follows:
0 to 10 hours: air volume 500m 3 /h～1200m 3 /h；
11-20 hours: air volume 1200m 3 /h～2000m 3 /h；
21-30 hours: air volume 2000m 3 /h～2800m 3 /h；
31-40 hours: air volume 2800m 3 /h～3600m 3 /h；
41-50 hours: air volume 3600m 3 /h～4200m 3 /h；
51 hours to tank discharge: air volume 4200m 3 /h～3800m 3 /h。
The fermentation medium in the step A consists of the following raw materials in percentage by mass:
3% of starch sugar; sodium nitrate 0.15%; corn steep liquor 0.2%; yeast extract 0.03%; potassium dihydrogen phosphate 0.1%; dipotassium hydrogen phosphate 0.1%; 0.015% of magnesium sulfate; 0.03% of defoaming agent; the balance being sterile water.
In the step A, latin name is Xanthomonas campestris, and after the xanthomonas campestris with the preservation number of CGMCC No.17845 is subjected to expansion culture, the xanthomonas campestris is subjected to seed liquid: fermentation medium = 5% inoculum size seed solution was inoculated into sterilized fermentation medium.
The step B comprises the following process steps:
b1, enzymolysis reaction: regulating the pH of the prepared fermentation liquor, adding alkaline protease for enzymolysis reaction, regulating the pH after the reaction is finished, and adding lysozyme for enzymolysis reaction;
b2, inactivating the enzyme preparation: adjusting the pH value of the enzymolysis liquid after the enzymolysis reaction in the step B to be 1.5, heating to 80 ℃ and preserving heat for 30 minutes;
b3, decoloring and filtering: cooling the reaction liquid prepared in the step C to 75 ℃, regulating the pH value to be 6.5, decoloring, adding a filter aid into the decolored reaction liquid, filtering, and collecting filtrate;
b4, shearing and ultrafiltering: and D, heating the filtrate prepared in the step D, homogenizing the heated reaction liquid under high pressure, and then performing ultrafiltration concentration to reach the solid content of 8%.
The alkaline protease treatment condition in the step B1 is that 0.02 percent of alkaline protease is added according to the amount of fermentation liquor, and the reaction is carried out for 3 hours under the heat preservation condition; the lysozyme treatment condition is that 0.01 percent of lysozyme is added according to the amount of the fermentation liquid, and the temperature is kept for 2 hours for reaction.
The enzyme activity of the alkaline protease is 20 mu/mg, the enzymolysis reaction temperature of the alkaline protease is 50-60 ℃, and the pH=9.5-10.5; the lysozyme adopts microbial cell wall hydrolase-glycoside hydrolase, the enzyme activity is 2 mu/mg, the temperature of the enzymolysis reaction of the lysozyme is 30-40 ℃, and the pH value is 4.5-6.5.
In the step B1, 10% NaOH solution or KOH solution with mass percent concentration is selected as pH value in the enzymolysis reaction of alkaline protease; the pH is adjusted by 10% acid solution in the enzymolysis reaction of lysozyme, and the acid is one of hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid and nitric acid.
In step B2, the ph=1.5 is adjusted with a 10% acid solution, wherein the acid is hydrochloric acid, sulfuric acid or nitric acid.
In the step B3, 10% NaOH solution or KOH solution with mass percentage concentration is used to adjust ph=6.5; decolorizing with 1-3% active carbon for 30-40 min; adding filter aid accounting for 1% of the total volume, filtering, and collecting filtrate; the filter aid adopts a mixture of diatomite and pearl salt, and the diatomite: mass ratio of pearl salt=2 to 4:6 to 8.
In the step B4, the temperature of the filtrate is raised to 70 ℃ and the filtrate is homogenized under the condition of 30 Mpa; the ultrafiltration condition is that the molecular weight cut-off is 50000 daltons, the ultrafiltration pressure is 0.2 MPa-0.3 MPa, and the concentration is carried out until the solid content is 8% -12%.
The method also comprises a step B5, wherein the process conditions of the step are as follows:
and B5, spray drying or alcohol extraction, drying and crushing the reaction liquid subjected to ultrafiltration concentration in the step B4 to prepare the solid low-viscosity xanthan gum.
Example 2
This embodiment differs from embodiment 1 in that:
the application of Xanthomonas campestris in preparing the low-viscosity xanthan gum comprises the following steps:
A. the Latin name is Xanthomonas campestris, and the preservation number is CGMCC No.17845, after the xanthomonas campestris is subjected to expansion culture, the xanthomonas campestris is subjected to seed liquid: inoculating the seed solution into a sterilized fermentation medium containing a carbon source, a nitrogen source and nutrient elements, and culturing under aeration culture to prepare a fermentation broth, wherein the fermentation medium is an inoculum size of 5% -15%;
the fermentation medium consists of the following raw materials in percentage by mass:
starch sugar 5%; sodium nitrate 0.3%; corn steep liquor 0.3%; yeast extract 0.06%; potassium dihydrogen phosphate 0.2%; dipotassium hydrogen phosphate 0.2%; magnesium sulfate 0.025%; 0.05% of defoaming agent; the balance being sterile water;
B. and C, collecting the low-viscosity xanthan gum from the fermentation liquor prepared in the step A, wherein the method comprises the following process steps of:
b1, enzymolysis reaction: regulating the pH of the prepared fermentation liquor, adding alkaline protease for enzymolysis reaction, regulating the pH after the reaction is finished, and adding lysozyme for enzymolysis reaction; the alkaline protease treatment condition is that 0.1% alkaline protease with the enzyme activity of 20 mu/mg is added according to the amount of fermentation broth, the temperature is 50-60 ℃ and the pH=9.5-10.5 is kept for reaction for 5 hours, and 10-20% NaOH solution or KOH solution with the mass percent concentration is used for adjusting the pH; the lysozyme treatment condition is that 0.05 percent of lysozyme is added according to the amount of the fermentation broth, and the temperature is kept for reaction for 4 hours; the lysozyme adopts microbial cell wall hydrolase-glycoside hydrolase, the enzyme activity is 2 mu/mg, the enzymolysis reaction temperature of the lysozyme is 30-40 ℃, the pH value is 4.5-6.5, the pH value is adjusted by using 20% acid solution, and the acid is one of hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid and nitric acid;
b2, inactivating the enzyme preparation: adjusting the pH value of the enzymolysis liquid after the enzymolysis reaction in the step B to be 3.0, heating to 95 ℃, and preserving heat for 60 minutes, wherein the pH value is adjusted by using a 20% acid solution, and the acid is hydrochloric acid, sulfuric acid or nitric acid;
b3, decoloring and filtering: cooling the reaction liquid prepared in the step C to 85 ℃, adjusting the pH value to be 7.5, and decoloring by adopting 1-3 per mill of activated carbon for 30-40 minutes; adding filter aid accounting for 1-2% of the total volume, filtering, and collecting filtrate; the filter aid adopts a mixture of diatomite and pearl salt, and the diatomite: mass ratio of pearl salt=2 to 4:6 to 8; adjusting the pH by using a NaOH solution or a KOH solution with the mass percent concentration of 20%;
b4, shearing and ultrafiltering: heating the filtrate prepared in the step D to 80 ℃, and homogenizing under the condition of 30 Mpa; concentrating the homogenized reactant by ultrafiltration until the solid content is 8% -12%, wherein the ultrafiltration condition is that the molecular weight cut-off is 50000 daltons, and the ultrafiltration pressure is 0.2-0.3 MPa.
The remainder was the same as in example 1.
Example 3
This embodiment differs from embodiment 1 in that:
the application of Xanthomonas campestris in preparing the low-viscosity xanthan gum comprises the following steps:
A. the Latin name is Xanthomonas campestris, and the preservation number is CGMCC No.17845, after the xanthomonas campestris is subjected to expansion culture, the xanthomonas campestris is subjected to seed liquid: inoculating the seed solution into a sterilized fermentation medium containing a carbon source, a nitrogen source and nutrient elements, and culturing under aeration culture to prepare a fermentation broth, wherein the fermentation medium is an inoculum size of 5% -15%;
the fermentation medium consists of the following raw materials in percentage by mass:
starch sugar 4%; sodium nitrate 0.2%; corn steep liquor 0.25%; yeast extract 0.04%; 0.15% of potassium dihydrogen phosphate; 0.15% of dipotassium hydrogen phosphate; 0.02% of magnesium sulfate; 0.04% of defoaming agent; the balance being sterile water;
B. and C, collecting the low-viscosity xanthan gum from the fermentation liquor prepared in the step A, wherein the method comprises the following process steps of:
b1, enzymolysis reaction: regulating the pH of the prepared fermentation liquor, adding alkaline protease for enzymolysis reaction, regulating the pH after the reaction is finished, and adding lysozyme for enzymolysis reaction; the alkaline protease treatment condition is that 0.06% alkaline protease with the enzyme activity of 20 mu/mg is added according to the amount of fermentation broth, the temperature is 50-60 ℃ and the pH=9.5-10.5 is kept for 4 hours for reaction, and 15% NaOH solution or KOH solution with the mass percent concentration is used for adjusting the pH; the lysozyme treatment condition is that 0.03 percent of lysozyme is added according to the amount of the fermentation broth, and the temperature is kept for reaction for 3 hours; the lysozyme adopts microbial cell wall hydrolase-glycoside hydrolase, the enzyme activity is 2 mu/mg, the enzymolysis reaction temperature of the lysozyme is 30-40 ℃, the pH value is 4.5-6.5, the pH value is adjusted by using 15% acid solution, and the acid is one of hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid and nitric acid;
b2, inactivating the enzyme preparation: adjusting the pH value of the enzymolysis liquid after the enzymolysis reaction in the step B to be 2.5, heating to 90 ℃, preserving heat for 45 minutes, and adjusting the pH value by using a 15% acid solution, wherein the acid is hydrochloric acid, sulfuric acid or nitric acid;
b3, decoloring and filtering: cooling the reaction liquid prepared in the step C to 80 ℃, adjusting the pH value to be 7.0, and decoloring by adopting 1-3 per mill of activated carbon for 30-40 minutes; adding filter aid accounting for 1-2% of the total volume, filtering, and collecting filtrate; the filter aid adopts a mixture of diatomite and pearl salt, and the diatomite: mass ratio of pearl salt=2 to 4:6 to 8; adjusting the pH by using 15% NaOH solution or KOH solution with mass percent concentration;
b4, shearing and ultrafiltering: heating the filtrate prepared in the step D to 75 ℃, and homogenizing under the condition of 30 Mpa; concentrating the homogenized reactant by ultrafiltration until the solid content is 8% -12%, wherein the ultrafiltration condition is that the molecular weight cut-off is 50000 daltons, and the ultrafiltration pressure is 0.2-0.3 MPa.
The remainder was the same as in example 1.
Example 4
This embodiment differs from embodiment 1 in that:
the application of Xanthomonas campestris in preparing the low-viscosity xanthan gum comprises the following steps:
A. the Latin name is Xanthomonas campestris, and the preservation number is CGMCC No.17845, after the xanthomonas campestris is subjected to expansion culture, the xanthomonas campestris is subjected to seed liquid: inoculating the seed solution into a sterilized fermentation medium containing a carbon source, a nitrogen source and nutrient elements, and culturing under aeration culture to prepare a fermentation broth, wherein the fermentation medium is an inoculum size of 5% -15%;
the fermentation medium consists of the following raw materials in percentage by mass:
equal amounts of starch sugar and sucrose 4.5%; sodium nitrate 0.25%; corn steep liquor 0.25%; yeast extract 0.05%; 0.18% of potassium dihydrogen phosphate; 0.18% of dipotassium hydrogen phosphate; magnesium sulfate 0.022%; 0.045% of defoaming agent; the balance being sterile water;
B. and C, collecting the low-viscosity xanthan gum from the fermentation liquor prepared in the step A, wherein the method comprises the following process steps of:
b1, enzymolysis reaction: regulating the pH of the prepared fermentation liquor, adding alkaline protease for enzymolysis reaction, regulating the pH after the reaction is finished, and adding lysozyme for enzymolysis reaction; the alkaline protease treatment condition is that 0.08% alkaline protease with the enzyme activity of 20 mu/mg is added according to the amount of fermentation broth, the temperature is 50-60 ℃ and the pH=9.5-10.5 is kept for 4.5 hours, and 18% NaOH solution or KOH solution with the mass percent concentration is used for adjusting the pH; the lysozyme treatment condition is that 0.01 to 0.05 percent of lysozyme is added according to the amount of fermentation liquor, and the temperature is kept for 3.5 hours; the lysozyme adopts microbial cell wall hydrolase-glycoside hydrolase, the enzyme activity is 2 mu/mg, the enzymolysis reaction temperature of the lysozyme is 30-40 ℃, the pH value is 4.5-6.5, 18% acid solution is used for regulating the pH value, and the acid is one of hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid and nitric acid;
b2, inactivating the enzyme preparation: adjusting the pH value of the enzymolysis liquid after the enzymolysis reaction in the step B to be 2.5, heating to 92 ℃, preserving heat for 55 minutes, and adjusting the pH value by using 18% acid solution, wherein the acid is hydrochloric acid, sulfuric acid or nitric acid;
b3, decoloring and filtering: cooling the reaction liquid prepared in the step C to 82 ℃, adjusting the pH=7.2, and decoloring by adopting 1-3 per mill of activated carbon for 30-40 minutes; adding filter aid accounting for 1-2% of the total volume, filtering, and collecting filtrate; the filter aid adopts a mixture of diatomite and pearl salt, and the diatomite: mass ratio of pearl salt=2 to 4:6 to 8; 10-20% NaOH solution or KOH solution with the mass percent concentration is used for regulating the pH value;
b4, shearing and ultrafiltering: heating the filtrate prepared in the step D to 70-80 ℃ and homogenizing under the condition of 30 Mpa; concentrating the homogenized reactant by ultrafiltration until the solid content is 8% -12%, wherein the ultrafiltration condition is that the molecular weight cut-off is 50000 daltons, and the ultrafiltration pressure is 0.2-0.3 MPa.
The remainder was the same as in example 1.
Example 5
This embodiment differs from embodiment 1 in that:
the application of Xanthomonas campestris in preparing the low-viscosity xanthan gum comprises the following steps:
A. the Latin name is Xanthomonas campestris, and the preservation number is CGMCC No.17845, after the xanthomonas campestris is subjected to expansion culture, the xanthomonas campestris is subjected to seed liquid: inoculating the seed solution into a sterilized fermentation medium containing a carbon source, a nitrogen source and nutrient elements, and culturing under aeration culture to prepare a fermentation broth, wherein the fermentation medium is an inoculum size of 5% -15%;
the fermentation medium consists of the following raw materials in percentage by mass:
3.5% of starch sugar and sucrose in a mass ratio of 3:7; sodium nitrate 0.18%; corn steep liquor 0.22%; yeast extract 0.035%; potassium dihydrogen phosphate 0.12%; 0.12% of dipotassium hydrogen phosphate; magnesium sulfate 0.018%; 0.035% of defoamer; the balance being sterile water;
B. and C, collecting the low-viscosity xanthan gum from the fermentation liquor prepared in the step A, wherein the method comprises the following process steps of:
b1, enzymolysis reaction: regulating the pH of the prepared fermentation liquor, adding alkaline protease for enzymolysis reaction, regulating the pH after the reaction is finished, and adding lysozyme for enzymolysis reaction; the alkaline protease treatment condition is that 0.04 percent of alkaline protease with the enzyme activity of 20 mu/mg is added according to the amount of fermentation broth, the temperature is 50-60 ℃ and the pH=9.5-10.5 is kept for 3.5 hours, and the pH is adjusted by using 12 percent NaOH solution or KOH solution with the mass percent concentration; the lysozyme treatment condition is that 0.01 to 0.05 percent of lysozyme is added according to the amount of fermentation liquor, and the temperature is kept for 2.5 hours; the lysozyme adopts microbial cell wall hydrolase-glycoside hydrolase, the enzyme activity is 2 mu/mg, the enzymolysis reaction temperature of the lysozyme is 30-40 ℃, the pH value is 4.5-6.5, the pH value is regulated to be 12% of acid solution, and the acid is one of hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid and nitric acid;
b2, inactivating the enzyme preparation: adjusting the pH value of the enzymolysis liquid after the enzymolysis reaction in the step B to be 1.5, heating to 86 ℃, preserving heat for 35 minutes, and adjusting the pH value by using a 12% acid solution, wherein the acid is hydrochloric acid, sulfuric acid or nitric acid;
b3, decoloring and filtering: cooling the reaction solution prepared in the step C to 78 ℃, adjusting the pH value to be 6.8, and decoloring by adopting 1-3 per mill of activated carbon for 30-40 minutes; adding filter aid accounting for 1-2% of the total volume, filtering, and collecting filtrate; the filter aid adopts a mixture of diatomite and pearl salt, and the diatomite: mass ratio of pearl salt=2 to 4:6 to 8; adjusting the pH by using a 12% NaOH solution or KOH solution with the mass percentage concentration;
b4, shearing and ultrafiltering: heating the filtrate prepared in the step D to 72 ℃, and homogenizing under the condition of 30 Mpa; concentrating the homogenized reactant by ultrafiltration until the solid content is 8% -12%, wherein the ultrafiltration condition is that the molecular weight cut-off is 50000 daltons, and the ultrafiltration pressure is 0.2-0.3 MPa.
The remainder was the same as in example 1.
The properties of the products prepared in the examples above are shown in the following table.
Claims (16)
1. The Xanthomonas campestris is characterized in that the Latin name of the strain is Xanthomonas camp estris, and the preservation number is CGMCC No.17845.
2. Use of xanthomonas campestris according to claim 1 in the preparation of a low viscosity xanthan gum.
3. The application according to claim 2, characterized in that the application comprises the steps of:
A. inoculating Latin name Xanthomonas campestris and accession number CGMCC No.17845 Xanthomonas campestris into fermentation medium containing carbon source, nitrogen source and nutrient element, and culturing under aeration to obtain fermentation broth;
B. and C, collecting the low-viscosity xanthan gum from the fermentation liquor prepared in the step A.
4. The use according to claim 3, wherein the aeration rate in the aeration culture in step A is 500m 3 /h～4200m 3 /h。
5. The use according to claim 3 or 4, wherein the ventilation in step a is:
0 to 10 hours: air volume 500m 3 /h～1200m 3 /h；
11-20 hours: air volume 1200m 3 /h～2000m 3 /h；
21-30 hours: air volume 2000m 3 /h～2800m 3 /h；
31-40 hours: air volume 2800m 3 /h～3600m 3 /h；
41-50 hours: air volume 3600m 3 /h～4200m 3 /h；
51 hours to tank discharge: air volume 4200m 3 /h～3800m 3 /h。
6. The method according to claim 3, wherein the fermentation medium in the step A uses sucrose and/or starch sugar as a carbon source and inorganic and organic small molecule nitrogen sources as nitrogen sources.
7. Use according to claim 3, characterized in that the fermentation medium in step a consists of the following raw materials in mass percent:
3% -5% of starch sugar and/or sucrose; sodium nitrate 0.15-0.3%; corn steep liquor 0.2% -0.3%; yeast extract 0.03-0.06%; 0.1 to 0.2 percent of monopotassium phosphate; 0.1 to 0.2 percent of dipotassium hydrogen phosphate; 0.015 to 0.025 percent of magnesium sulfate; 0.03 to 0.05 percent of defoaming agent; the balance being sterile water.
8. The application of claim 3, wherein in the step A, latin name is Xanthomonas campestris, and the preservation number is CGMCC No.17845, after the Xanthomonas campestris is subjected to expansion culture, the method comprises the following steps: fermentation medium = 5% -15% inoculum size seed fluid is inoculated into sterilized fermentation medium.
9. A use according to claim 3, characterized in that said step B comprises the following process steps:
b1, enzymolysis reaction: regulating the pH of the prepared fermentation liquor, adding alkaline protease for enzymolysis reaction, regulating the pH after the reaction is finished, and adding lysozyme for enzymolysis reaction;
b2, inactivating the enzyme preparation: regulating the pH value of the enzymolysis liquid after the enzymolysis reaction in the step B to be 1.5-3.0, heating to 80-95 ℃ and preserving heat for 30-60 minutes;
b3, decoloring and filtering: cooling the reaction liquid prepared in the step C to 75-85 ℃, regulating the pH value to be 6.5-7.5, decoloring, adding a filter aid into the decolored reaction liquid, filtering, and collecting filtrate;
b4, shearing and ultrafiltering: and D, heating the filtrate prepared in the step D, homogenizing the heated reaction liquid under high pressure, and then performing ultrafiltration concentration until the solid content is 8-12%.
10. The use according to claim 9, characterized in that the alkaline protease treatment conditions in step B1 are that 0.02% -0.1% alkaline protease is added according to the amount of the fermentation broth, and the reaction is carried out for 3-5 hours; the lysozyme treatment condition is that 0.01 to 0.05 percent of lysozyme is added according to the amount of the fermentation liquid, and the temperature is kept for 2 to 4 hours.
11. The use according to claim 10, characterized in that the enzymatic activity of the alkaline protease is 20 u/mg, the temperature of the enzymatic hydrolysis of the alkaline protease is 50-60 ℃, pH = 9.5-10.5; the lysozyme adopts microbial cell wall hydrolase-glycoside hydrolase, the enzyme activity is 2 mu/mg, the temperature of the enzymolysis reaction of the lysozyme is 30-40 ℃, and the pH value is 4.5-6.5.
12. The use according to claim 11, characterized in that in step B1, the alkaline protease enzymatic hydrolysis is performed with a pH-adjusting solution of 10% to 20% NaOH or KOH; the pH is adjusted by 10-20% acid solution in the enzymolysis reaction of lysozyme, and the acid is one of hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid and nitric acid.
13. The use according to claim 9, characterized in that in step B2 the ph=1.5-3.0 is adjusted with a 10-20% acid solution, said acid being selected from hydrochloric acid, sulfuric acid or nitric acid.
14. Use according to claim 9, characterized in that in step B3 the ph=6.5-7.5 is adjusted with 10-20% NaOH solution or KOH solution in mass percent; decolorizing with 1-3% active carbon for 30-40 min; adding filter aid accounting for 1-2% of the total volume, filtering, and collecting filtrate; the filter aid adopts a mixture of diatomite and pearl salt, and the diatomite: mass ratio of pearl salt=2 to 4:6 to 8.
15. The use according to claim 9, characterized in that in step B4 the filtrate is heated to 70-80 ℃ and homogenized at a pressure of 30 Mpa; the ultrafiltration condition is that the molecular weight cut-off is 50000 daltons, the ultrafiltration pressure is 0.2-0.3 MPa, and the concentration is carried out until the solid content is 8-12%.
16. The use according to claim 9, further comprising a step B5, the process conditions of which are as follows:
and B5, spray drying or alcohol extraction, drying and crushing the reaction liquid subjected to ultrafiltration concentration in the step B4 to prepare the solid low-viscosity xanthan gum.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117568233A (en) * | 2023-12-22 | 2024-02-20 | 内蒙古工业大学 | Preparation method of xanthan gum capable of reducing pyruvic acid content and improving salt tolerance |
| CN117568232A (en) * | 2023-12-21 | 2024-02-20 | 内蒙古工业大学 | Xanthomonas campestris capable of producing high-yield temperature-resistant instant xanthan gum and application thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117568232A (en) * | 2023-12-21 | 2024-02-20 | 内蒙古工业大学 | Xanthomonas campestris capable of producing high-yield temperature-resistant instant xanthan gum and application thereof |
| CN117568233A (en) * | 2023-12-22 | 2024-02-20 | 内蒙古工业大学 | Preparation method of xanthan gum capable of reducing pyruvic acid content and improving salt tolerance |
| CN117568233B (en) * | 2023-12-22 | 2024-03-22 | 内蒙古工业大学 | Preparation method of xanthan gum capable of reducing pyruvic acid content and improving salt tolerance |
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