Method for producing gellan gum by fermentation of bagasse and cold-gelled gum
After treating bagasse powder with specific enzymes, it is used as a carbon source and sugar source of Sphingosine Monassia to ferment and produce gellan gum, which solves the problems of low productivity and high cost of microbial gellan gum in the prior art, and realizes effective utilization of resources and increase gellan gum production.
Patent Information
- Application Number
- CN202411944709.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-27
- Publication Date
- 2025-04-04
AI Technical Summary
The existing microbial gellan gum has low productivity and high production costs, which makes it economically unfeasible, and agricultural and forestry waste such as bagasse have not been effectively utilized.
Gelectyl gelatin is fermented and produced by fermenting bagasse powder with cellulase, amylase and saccharase as a carbon source and sugar source of Sphingosine Monassia. The method includes preparing bagasse powder, adding culture medium after treatment, fermenting, heating, centrifugation and washing to obtain gellan gum.
This method not only effectively utilizes waste such as sugarcane bagasse, reduces the fermentation cost of gellan gum, but also increases the yield and quality of gellan gum, and solves the problem of resource waste.
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Figure CN119753057A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of cold-setting gum production, and particularly relates to a method for producing gellan gum by fermenting bagasse and the cold-setting gum. Background Art
[0002] Gellan gum is a new type of microbial polysaccharide produced by Sphingomonas paucimobilis. It has excellent properties such as low dosage, high transparency, acid resistance, high temperature resistance, enzyme resistance, and good compatibility. It has a wide range of uses in the food, pharmaceutical, and chemical industries. However, due to the low productivity and high production cost of microbial gellan gum, the production of microbial gellan gum is not economically feasible. The use of low-cost carbon sources can help reduce the cost of gellan gum.
[0003] Bagasse is the main byproduct of the sugarcane sugar industry, accounting for 24%-27%. The fibrous residue after the sugarcane is crushed and squeezed to extract the juice is a solid waste of the sugar industry and a renewable resource that can be utilized. The main components of bagasse are 42-50% cellulose, 25-30% hemicellulose, 20-25% lignin and other ash. Its composition varies depending on the type, quality, growth conditions and squeezing technology of sugarcane. my country is a big agricultural country with abundant agricultural and forestry wastes rich in cellulose and hemicellulose, such as bagasse, corn cobs, wheat bran, and straw, which have long been neglected and not fully developed and utilized. Less than 2% of them are used for industrial combustion, and most of them are discarded in the fields or directly burned, causing environmental pollution. Summary of the invention
[0004] The invention aims to provide a method for producing gellan gum by fermenting bagasse and the gellan gum.
[0005] The first aspect of the present invention provides a method for producing gellan gum by fermenting bagasse, comprising:
[0006] preparing bagasse powder;
[0007] Add bagasse powder to the cellulase solution and place in a 50°C water bath for 12-48h;
[0008] Add amylase and saccharifying enzyme, and place in a 70℃ water bath for 1-10h;
[0009] Boil for 10 min and then dry to obtain bagasse extract;
[0010] Prepare a culture medium containing bagasse extract, adjust to pH=7.0, and sterilize;
[0011] The cultured Sphingomonas bacterial liquid is inoculated into the prepared culture medium at a 10% inoculation rate for fermentation;
[0012] After fermentation, the fermentation liquid was heated to 90°C with stirring, maintained for 0.5 h, cooled naturally to room temperature, and centrifuged at 10,000 rpm for 1 h;
[0013] The mixture was washed three times with washing liquid, the supernatant was removed, and the precipitate was dried at 50°C to obtain gellan gum.
[0014] In one embodiment of the present application, the method for preparing bagasse powder comprises:
[0015] The bagasse was naturally air-dried and then crushed, passed through a 100-mesh sieve, dried at 80°C to constant weight, and stored in a sealed container at room temperature.
[0016] In one embodiment of the present application, the concentration of the cellulase solution is 5-20wt%;
[0017] The weight-volume ratio of the bagasse powder to the cellulase solution is 1:100-1:500 g / ml.
[0018] In one embodiment of the present application, the final concentration of the amylase is 0.2-2 wt %; the final concentration of the saccharifying enzyme is 0.5-5 wt %.
[0019] In one embodiment of the present application, the raw materials of the culture medium include, in final concentrations: 10 g / L Na2HPO4, 1 g / L K2SO4, 1 g / L NaCl, 0.15 g / L (NH4)2SO4, 0.01 g / L CaCl2.2H2O, 0.2 g / L MgSO4.7H2O, 0.001 g / L FeSO4.7H2O, 0.5 g / L yeast extract, and 30-100 g / L bagasse extract; the above raw materials are dissolved in water when preparing the culture medium.
[0020] In one embodiment of the present application, the total number of bacterial colonies contained in each milliliter of the Sphingomonas bacterial solution is 1*10 8 CFU / mL.
[0021] In one embodiment of the present application, the washing liquid is a solution of isopropyl alcohol: water = 1:3 in volume ratio.
[0022] Accordingly, a second aspect of the present invention provides a gellan gum prepared by the method for producing gellan gum by fermenting bagasse as described above.
[0023] The beneficial effect of the present invention is that the method for producing gellan gum by fermenting bagasse of the present invention adopts the waste resource product bagasse, and utilizes the treated bagasse as the carbon source and sugar source for bacterial fermentation, which can not only solve the waste of resources, but also reduce the fermentation cost of gellan gum.
[0024] Other features and advantages of the present invention will be described in the following description, and partly become apparent from the description, or understood by practicing the present invention. The purpose and other advantages of the present invention are realized and obtained by the structures particularly pointed out in the description and the drawings.
[0025] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, preferred embodiments are given below and described in detail with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS
[0026] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.
[0027] Figure 1 It is a schematic diagram of the test results of the fermentation broth viscosity of the embodiment of the present invention and the comparative example 1;
[0028] Figure 2 Schematic diagram comparing the gellan gum yields of the embodiments of the present invention and the comparative example 1. DETAILED DESCRIPTION
[0029] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in conjunction with the accompanying drawings. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0030] Example 1
[0031] (1) The bagasse was naturally air-dried and then crushed, passed through a 100-mesh sieve, dried at 80°C to constant weight, and stored in a sealed container at room temperature.
[0032] (2) Add 5 wt% cellulase and incubate in a 50°C water bath for 48 h.
[0033] (3) Add amylase with a final concentration of 0.2 wt% and saccharifying enzyme with a final concentration of 0.5 wt%, place in a 70°C water bath for 10 h, boil for 10 min, and dry.
[0034] (4) Prepare a culture medium, wherein the raw materials of the culture medium include, in final concentrations, 10 g / L Na2HPO4, 1 g / L K2SO4, 1 g / L NaCl, 0.15 g / L (NH4)2SO4, 0.01 g / L CaCl2.2H2O, 0.2 g / LMgSO4.7H2O, 0.001 g / L FeSO4.7H2O, 0.5 g / L yeast extract, and 30 g / L bagasse extract; dissolve the above raw materials in water, adjust the pH to 7.0, and sterilize.
[0035] (5) The cultured Sphingomonas (1*10 8 CFU / mL) bacterial solution was inoculated into the prepared culture medium at a 10% inoculation rate for fermentation.
[0036] (6) After fermentation, the fermentation liquid was heated to 90°C with stirring for 0.5 h, cooled naturally to room temperature, and centrifuged at 10,000 rpm for 1 h.
[0037] (7) Wash three times with a solution of isopropanol:water = 1:3, remove the supernatant, and dry the precipitate at 50°C to obtain gellan gum.
[0038] Example 2
[0039] (1) The bagasse was naturally air-dried and then crushed, passed through a 100-mesh sieve, dried at 80°C to constant weight, and stored in a sealed container at room temperature.
[0040] (2) Add 10 wt% cellulase and place in a 50°C water bath for 24 h.
[0041] (3) Add amylase with a final concentration of 0.5 wt% and saccharifying enzyme with a final concentration of 2 wt% to the above solution, place in a 70°C water bath for 5 h, boil for 10 min, and dry.
[0042] (4) Prepare a culture medium, wherein the raw materials of the culture medium include, in final concentrations, 10 g / L Na2HPO4, 1 g / L K2SO4, 1 g / L NaCl, 0.15 g / L (NH4)2SO4, 0.01 g / L CaCl2.2H2O, 0.2 g / LMgSO4.7H2O, 0.001 g / L FeSO4.7H2O, 0.5 g / L yeast extract, and 30 g / L bagasse extract; dissolve the above raw materials in water, adjust the pH to 7.0, and sterilize.
[0043] (5) The cultured Sphingomonas (1*10 8 CFU / mL) bacterial solution was inoculated into the prepared culture medium at a 10% inoculation rate for fermentation.
[0044] (6) After fermentation, the fermentation liquid was heated to 90°C with stirring for 0.5 h, cooled naturally to room temperature, and centrifuged at 10,000 rpm for 1 h.
[0045] (7) Wash three times with a solution of isopropanol:water = 1:3, remove the supernatant, and dry the precipitate at 50°C to obtain gellan gum.
[0046] Example 3
[0047] (1) The bagasse was naturally air-dried and then crushed, passed through a 100-mesh sieve, dried at 80°C to constant weight, and stored in a sealed container at room temperature.
[0048] (2) Add 15 wt% cellulase and incubate in a 50°C water bath for 24 h.
[0049] (3) Add 1 wt% of amylase and 1 wt% of saccharifying enzyme to the above solution, place in a 70°C water bath for 5 h, boil for 10 min, and dry.
[0050] (4) Prepare a culture medium, wherein the raw materials of the culture medium include, in final concentrations, 10 g / L Na2HPO4, 1 g / L K2SO4, 1 g / L NaCl, 0.15 g / L (NH4)2SO4, 0.01 g / L CaCl2.2H2O, 0.2 g / LMgSO4.7H2O, 0.001 g / L FeSO4.7H2O, 0.5 g / L yeast extract, and 30 g / L bagasse extract; dissolve the above raw materials in water, adjust the pH to 7.0, and sterilize.
[0051] (5) The cultured Sphingomonas (1*10 8 CFU / mL) bacterial solution was inoculated into the prepared culture medium at a 10% inoculation rate for fermentation.
[0052] (6) After fermentation, the fermentation liquid was heated to 90°C with stirring for 0.5 h, cooled naturally to room temperature, and centrifuged at 10,000 rpm for 1 h.
[0053] (7) Wash three times with a solution of isopropanol:water = 1:3, remove the supernatant, and dry the precipitate at 50°C to obtain gellan gum.
[0054] Example 4
[0055] (1) The bagasse was naturally air-dried and then crushed, passed through a 100-mesh sieve, dried at 80°C to constant weight, and stored in a sealed container at room temperature.
[0056] (2) Add 20 wt% cellulase and place in a 50°C water bath for 12-48 hours.
[0057] (3) Add 1 wt% of amylase and 3 wt% of saccharifying enzyme to the above solution, place in a 70°C water bath for 4 h, boil for 10 min, and dry.
[0058] (4) Prepare a culture medium, wherein the raw materials of the culture medium include, in final concentrations, 10 g / L Na2HPO4, 1 g / L K2SO4, 1 g / L NaCl, 0.15 g / L (NH4)2SO4, 0.01 g / L CaCl2.2H2O, 0.2 g / LMgSO4.7H2O, 0.001 g / L FeSO4.7H2O, 0.5 g / L yeast extract, and 50 g / L bagasse extract; dissolve the above raw materials in water, adjust the pH to 7.0, and sterilize.
[0059] (5) The cultured Sphingomonas (1*10 8 CFU / mL) bacterial solution was inoculated into the prepared culture medium at a 10% inoculation rate for fermentation.
[0060] (6) After fermentation, the fermentation liquid was heated to 90°C with stirring for 0.5 h, cooled naturally to room temperature, and centrifuged at 10,000 rpm for 1 h.
[0061] (7) Wash three times with a solution of isopropanol:water = 1:3, remove the supernatant, and dry the precipitate at 50°C to obtain gellan gum.
[0062] Example 5
[0063] (1) The bagasse was naturally air-dried and then crushed, passed through a 100-mesh sieve, dried at 80°C to constant weight, and stored in a sealed container at room temperature.
[0064] (2) Add 20 wt% cellulase and incubate in a 50°C water bath for 12 h.
[0065] (3) Add 2 wt% of amylase and 5 wt% of saccharifying enzyme to the above solution, place in a 70°C water bath for 1 hour, boil for 10 minutes, and dry.
[0066] (4) Prepare a culture medium, wherein the raw materials of the culture medium include, in final concentrations, 10 g / L Na2HPO4, 1 g / L K2SO4, 1 g / L NaCl, 0.15 g / L (NH4)2SO4, 0.01 g / L CaCl2.2H2O, 0.2 g / LMgSO4.7H2O, 0.001 g / L FeSO4.7H2O, 0.5 g / L yeast extract, and 100 g / L bagasse extract; dissolve the above raw materials in water, adjust the pH to 7.0, and sterilize.
[0067] (5) The cultured Sphingomonas (1*10 8 CFU / mL) bacterial solution was inoculated into the prepared culture medium at a 10% inoculation rate for fermentation.
[0068] (6) After fermentation, the fermentation liquid was heated to 90°C with stirring for 0.5 h, cooled naturally to room temperature, and centrifuged at 10,000 rpm for 1 h.
[0069] (7) Wash three times with a solution of isopropanol:water = 1:3, remove the supernatant, and dry the precipitate at 50°C to obtain gellan gum.
[0070] Comparative Example 1
[0071] (1) Prepare a culture medium, wherein the raw materials of the culture medium include, in final concentrations, 10 g / L Na2HPO4, 1 g / L K2SO4, 1 g / L NaCl, 0.15 g / L (NH4)2SO4, 0.01 g / L CaCl2.2H2O, 0.2 g / LMgSO4.7H2O, 0.001 g / L FeSO4.7H2O, and 0.5 g / L yeast extract; dissolve the above raw materials in water, adjust the pH to 7.0, and sterilize.
[0072] Yeast extract is a commonly used culture medium component that can be refined from yeast and used to prepare microbial culture medium to provide nitrogen source and B vitamins.
[0073] (2) The cultured Sphingomonas (1*10 8 CFU / mL) bacterial solution was inoculated into the prepared culture medium at a 10% inoculation rate for fermentation.
[0074] (3) After fermentation, the fermentation liquid was heated to 90°C with stirring for 0.5 h, cooled naturally to room temperature, and centrifuged at 10,000 rpm for 1 h.
[0075] (4) Washing the mixture three times with a solution of isopropanol:water = 1:3, removing the supernatant, and drying the precipitate at 50°C to obtain gellan gum.
[0076] Fermentation broth viscosity test:
[0077] 50 mL of fermentation liquid was taken out from Examples 1-5 and Comparative Example 1 at regular intervals and placed in a small beaker so that the end of the rotor was submerged by the fermentation liquid. The rotational viscometer NDJ-1 was started and the result was recorded after the reading stabilized after two rotations, which was the viscosity of the fermentation liquid. Figure 1 As shown, as the fermentation time increases, the viscosity of the nutrient solution in Examples 1-5 reaches the highest value at 24-26h, indicating that the gellan gum yield is the highest at this time. At the same time, the viscosity in Control Example 1 is much lower than that in Examples 1-5, indicating that bagasse extract plays an important role in the production of gellan gum.
[0078] Gellan gum production detection:
[0079] The gellan gum powder obtained in Examples 1-5 and Comparative Example 1 was weighed, and the gellan gum yield was calculated according to the following formula:
[0080] Gellan gum yield = powder weight / fermentation liquid volume;
[0081] The results are as follows Figure 2 As shown, the gellan gum produced by Examples 1-5 is much greater than that of Control Example 1, indicating that the sugarcane bagasse extract is helpful for bacterial biosynthesis of gellan gum. The gellan gum yield of Example 5 is the highest, which may be due to the addition of the most cellulase, which can process the sugarcane bagasse to generate a larger proportion of monosaccharides.
[0082] The various devices selected in this application (components whose specific structures are not described) are all universal standard parts or components known to technical personnel in this field, and their structures and principles can be known to technical personnel through technical manuals or through conventional experimental methods.
[0083] Based on the above ideal embodiments of the present invention, the relevant staff can make various changes and modifications without departing from the technical concept of the present invention through the above description. The technical scope of the present invention is not limited to the contents of the specification, and its technical scope must be determined according to the scope of the claims.
Claims
1. A method for producing gellan gum by fermenting bagasse, characterized in that: include: preparing bagasse powder; Add bagasse powder to the cellulase solution and place in a 50°C water bath for 12-48h; Add amylase and saccharifying enzyme, and place in a 70℃ water bath for 1-10h; Boil for 10 min and then dry to obtain bagasse extract; Prepare a culture medium containing bagasse extract, adjust to pH=7.0, and sterilize; The cultured Sphingomonas bacterial liquid is inoculated into the prepared culture medium at a 10% inoculation rate for fermentation; After fermentation, the fermentation liquid was heated to 90°C with stirring, maintained for 0.5 h, cooled naturally to room temperature, and centrifuged at 10,000 rpm for 1 h; The mixture was washed three times with washing liquid, the supernatant was removed, and the precipitate was dried at 50°C to obtain gellan gum.
2. The method for producing gellan gum by fermenting bagasse according to claim 1, characterized in that: The method for preparing bagasse powder comprises: The bagasse was naturally air-dried and then crushed, passed through a 100-mesh sieve, dried at 80°C to constant weight, and stored in a sealed container at room temperature.
3. The method for producing gellan gum by fermenting bagasse according to claim 1, characterized in that: The concentration of the cellulase solution is 5-20wt%; The weight-volume ratio of the bagasse powder to the cellulase solution is 1:100-1:500 g / ml.
4. The method for producing gellan gum by fermenting bagasse according to claim 1, characterized in that: The final concentration of the amylase is 0.2-2wt%; The final concentration of the saccharifying enzyme is 0.5-5wt%.
5. The method for producing gellan gum by fermenting bagasse according to claim 1, characterized in that: The raw materials of the culture medium include: 10g / L Na2HPO4, 1g / L K2SO4, 1g / L NaCl, 0.15g / L (NH4)2SO4, 0.01g / L CaCl2.2H2O, 0.2g / L MgSO4.7H2O, 0.001g / L FeSO4.7H2O, 0.5g / L yeast extract, and 30-100g / L bagasse extract.
6. The method for producing gellan gum by fermenting bagasse according to claim 1, characterized in that: The total number of bacterial colonies contained in each milliliter of the Sphingomonas bacterial solution is 1*10 8 CFU / mL.
7. The method for producing gellan gum by fermenting bagasse according to claim 1, characterized in that: The washing liquid is a solution of isopropanol:water=1:3 in volume ratio.
8. A gellan gum, characterized in that: The method for producing gellan gum by fermenting bagasse as described in any one of claims 1 to 7 is adopted.