CN116622794A - Method for preparing bacterial cellulose by using vinegar residues - Google Patents
Method for preparing bacterial cellulose by using vinegar residues Download PDFInfo
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- CN116622794A CN116622794A CN202310734777.XA CN202310734777A CN116622794A CN 116622794 A CN116622794 A CN 116622794A CN 202310734777 A CN202310734777 A CN 202310734777A CN 116622794 A CN116622794 A CN 116622794A
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- 239000000052 vinegar Substances 0.000 title claims abstract description 82
- 235000021419 vinegar Nutrition 0.000 title claims abstract description 82
- 229920002749 Bacterial cellulose Polymers 0.000 title claims abstract description 67
- 239000005016 bacterial cellulose Substances 0.000 title claims abstract description 67
- 238000000034 method Methods 0.000 title claims abstract description 32
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 54
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 33
- 238000001914 filtration Methods 0.000 claims abstract description 21
- 108010059892 Cellulase Proteins 0.000 claims abstract description 17
- 229940106157 cellulase Drugs 0.000 claims abstract description 17
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- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 39
- 239000000243 solution Substances 0.000 claims description 39
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- 239000011259 mixed solution Substances 0.000 claims description 14
- XPFJYKARVSSRHE-UHFFFAOYSA-K trisodium;2-hydroxypropane-1,2,3-tricarboxylate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].[Na+].OC(=O)CC(O)(C(O)=O)CC(O)=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O XPFJYKARVSSRHE-UHFFFAOYSA-K 0.000 claims description 10
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- 238000011081 inoculation Methods 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 abstract description 9
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- 238000012258 culturing Methods 0.000 description 5
- 239000002054 inoculum Substances 0.000 description 5
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 4
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- 239000007864 aqueous solution Substances 0.000 description 4
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 4
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 4
- 239000008367 deionised water Substances 0.000 description 4
- 229910021641 deionized water Inorganic materials 0.000 description 4
- 238000001784 detoxification Methods 0.000 description 4
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- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
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- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 2
- YLQBMQCUIZJEEH-UHFFFAOYSA-N Furan Chemical compound C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
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- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 2
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- 235000009025 Carya illinoensis Nutrition 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
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- 102000004190 Enzymes Human genes 0.000 description 1
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- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
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- 238000010306 acid treatment Methods 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
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- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000001027 hydrothermal synthesis Methods 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 239000002440 industrial waste Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P2201/00—Pretreatment of cellulosic or lignocellulosic material for subsequent enzymatic treatment or hydrolysis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P2203/00—Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Processing Of Solid Wastes (AREA)
Abstract
The invention discloses a method for preparing bacterial cellulose by using vinegar residues, which comprises the following steps: (1) taking vinegar residues, drying and crushing; (2) placing the vinegar residues in an acetic acid solution for acidolysis; filtering, and washing the acidolysis vinegar residue to neutrality with water; (3) Placing vinegar residues in water, regulating the pH value to 4.8-5.0, and adding cellulase for enzymolysis; filtering; (4) Mixing acidolysis solution and enzymolysis solution, and adding active carbon to detoxify and decolor; filtering and centrifuging to obtain a fermentation medium; (5) Sterilizing the fermentation medium, inoculating Kang Pucha bacteria, standing and fermenting at normal temperature for 7-14 days to obtain the bacterial cellulose membrane. The method for preparing the bacterial cellulose by using the vinegar residue improves the utilization value of the vinegar residue, reduces the pressure of the vinegar residue on the environment, reduces the preparation cost of the bacterial cellulose, provides a new raw material for the industrial production of the bacterial cellulose, and simultaneously provides possibility for the high-valued application of the bacterial fiber.
Description
Technical Field
The invention relates to a method for preparing bacterial cellulose by using vinegar residues, and belongs to the technical field of preparation of bacterial cellulose.
Background
Cellulose is a three-dimensional (3D) network polymer consisting of glucose units linked by beta-1, 4 glycosidic linkages. Each repeating glucose unit has three hydroxyl groups. Cellulose is one of the most abundant, most environmentally friendly natural biopolymers, which can be divided into two categories depending on the source: plant Cellulose (PC) and Bacterial Cellulose (BC). PC is mainly obtained from cotton, wood and algae. BC is mainly obtained from various strains such as Staphylococcus (glucoacetobacter), sarcoma (Sarcina), and aerobe (aerobe). Compared with PC, BC has the advantages of high purity, high strength, high crystallinity, good biocompatibility, large specific surface area and the like, so BC is widely applied to the fields of medicines, food packaging, paper products, cosmetics, electronic products and the like. However, the high production cost of BC has limited its extensive academic research and commercial use. The media cost is about 30% of the total BC production cost, and therefore, some industrial and agricultural waste has been tested as an alternative medium for BC production, including pecan hulls, potato peel waste, orange peel waste, and winery waste.
Vinegar residues are main byproducts generated by fermenting raw materials (rice, corn, sorghum, sweet potato, barley and the like) in the vinegar production process, and are generally buried and landfilled directly by manufacturers, but can pollute soil and underground water. The vinegar residue mainly comprises cellulose, hemicellulose and lignin, and no report on the preparation of BC by taking the vinegar residue as a culture medium is available at present.
Current academic research focuses on innovative and sustainable methods to recycle biomass residues. Hydrothermal techniques are commonly used to pretreat lignocellulosic feedstock, disrupt material structure, promote hemicellulose hydrolysis, and increase accessibility of cellulose. Inorganic acids (e.g., sulfuric acid, hydrochloric acid) have found wide application in hydrothermal processes due to their relatively high catabolic activity. However, the use of mineral acids has a number of disadvantages, including corrosion of the equipment and the generation of sewage.
Disclosure of Invention
Aiming at the prior art, the invention provides a method for preparing bacterial cellulose by using vinegar residues. According to the method for preparing bacterial cellulose by using vinegar residues, acetic acid acidolysis of the vinegar residues is adopted, hemicellulose is degraded into xylose and arabinose under the effect of acid treatment, and a small part of cellulose is converted into glucose; then adding cellulase for enzymolysis, and degrading the cellulose into glucose under the action of enzyme treatment; the vinegar residue after acidolysis and enzymolysis treatment can be used as a carbon source for bacterial proliferation and used for preparing bacterial cellulose.
The invention is realized by the following technical scheme:
a method for preparing bacterial cellulose by using vinegar residues, which comprises the following steps:
(1) Taking vinegar residues, drying and crushing;
(2) Placing the dried and crushed vinegar residues into an acetic acid solution, and acidolyzing for 30-90 minutes at the temperature of 100-180 ℃; filtering to obtain acidolysis solution and acidolysis vinegar residue, and washing acidolysis vinegar residue with water to neutrality;
(3) Placing the acidolysis vinegar residues in water or buffer solution, and regulating the pH to 4.8-5.0 to obtain to-be-enzymatically hydrolyzed solution; adding cellulase, and carrying out enzymolysis for 4-24 hours at 48-52 ℃; filtering to obtain enzymolysis liquid;
(4) Mixing the acidolysis solution and the enzymolysis solution to obtain a mixed solution, and adding activated carbon to detoxify and decolor; filtering and centrifuging to obtain a fermentation medium;
(5) Sterilizing the fermentation medium at 121 ℃ for 20 minutes, inoculating Kang Pucha bacteria (conventional strains existing in the prior art), standing at normal temperature, fermenting and culturing for 7-14 days to obtain the bacterial cellulose membrane.
Further, in the step (1), the drying mode is selected from a natural drying method, a forced air drying method and the like; the drying temperature is 60-80 ℃.
Further, in the step (1), the crushed materials are sieved by a sieve with 30 to 100 meshes.
Further, in the step (2), the mass ratio of the vinegar residue to the acetic acid is 1:10-20.
Further, in the step (2), the concentration of the acetic acid solution is 1% -5% (mass percentage).
Further, in the step (3), the buffer solution is a citric acid-sodium citrate buffer solution, and the mass ratio of the vinegar residue to water or the buffer solution is 1:10-25.
Further, in the step (3), the mass ratio of the cellulase to the liquid to be subjected to enzymolysis is 1:50-200.
Further, in the step (4), the mass ratio of the activated carbon to the mixed solution is 1:20-100.
Further, in the step (4), activated carbon is added, and then the mixture is stirred for 15 to 60 minutes at the temperature of between 25 and 100 ℃ to detoxify and decolor.
Further, in the step (5), the inoculation amount is 8-15% (volume percent).
Further, the method also comprises the following steps: (6) Purifying bacterial cellulose in sodium hydroxide solution to obtain a purified bacterial cellulose membrane; the concentration of the sodium hydroxide solution is 1-5% (mass percent), the purification temperature is 80-100 ℃ and the time is 30-60 minutes.
According to the method for preparing bacterial cellulose by using the vinegar residues, acetic acid is firstly used for acidolysis, hemicellulose in the vinegar residues can be converted into xylose and arabinose, a small amount of cellulose is converted into glucose, the reducing sugar content and the conversion rate are improved, meanwhile, the surface structure of the cellulose is fluffy, the cellulose is easier to be subjected to enzymolysis in the follow-up process, the using amount of cellulase is reduced, and the preparation cost of the bacterial cellulose is saved. Acetic acid is an organic acid and has low acidity and less damage to equipment than inorganic acids (sulfuric acid, hydrochloric acid). And then, performing enzymolysis by using cellulase, wherein the cellulose is thoroughly degraded by the vinegar residue under the action of the cellulase. And then, adsorbing impurities such as furan, acid-soluble lignin and the like in the acidolysis solution and the enzymolysis solution by using activated carbon to prepare a detoxified decoloration acidolysis and enzymolysis mixed solution, wherein the pH value of the mixed solution accords with the growth of bacterial cellulose, and the pH value is not required to be regulated in the fermentation culture process, so that the process flow is shortened. Then Kang Pucha bacteria are inoculated, bacterial cellulose membranes are synthesized by fermentation, and the cost of the culture medium is reduced and the equipment cost is reduced by normal-temperature static culture under the condition that a carbon source and a nitrogen source required by microorganism growth are not additionally added. And (3) performing impurity removal and sterilization treatment on the bacterial cellulose membrane in a sodium hydroxide solution to prepare the purified bacterial cellulose membrane. According to the invention, kang Pucha bacteria are utilized to prepare bacterial cellulose, and Kang Pucha bacteria can generate tea polyphenol (with antibacterial property) in the proliferation process, so that the bacterial cellulose also has antibacterial property.
According to the method for preparing the bacterial cellulose by using the vinegar residues, the renewable resources of the vinegar residues are converted into the microbial cellulose, so that the utilization value of the vinegar residues is improved, the pressure of the vinegar residues on the environment is reduced, meanwhile, the preparation cost of the bacterial cellulose is reduced, a new raw material is provided for the industrial production of the bacterial cellulose, and meanwhile, the possibility is provided for the high-valued application of the bacterial fibers.
Drawings
Fig. 1: SEM image of bacterial cellulose.
Fig. 2: diameter profile of bacterial cellulose.
Fig. 3: FTIR plot of bacterial cellulose.
Fig. 4: XRD pattern of bacterial cellulose.
Fig. 5: yield comparative schematic.
Detailed Description
The invention is further illustrated below with reference to examples. However, the scope of the present invention is not limited to the following examples. Those skilled in the art will appreciate that various changes and modifications can be made to the invention without departing from the spirit and scope thereof.
The instruments, reagents and materials used in the examples below are conventional instruments, reagents and materials known in the art and are commercially available. The experimental methods, detection methods, and the like in the examples described below are conventional experimental methods and detection methods known in the prior art unless otherwise specified.
Example 1 preparation of bacterial cellulose Using Vinegar residue
The method comprises the following steps:
(1) And (3) carrying out forced air drying on the vinegar residues, crushing, and sieving the crushed vinegar residues with a 100-mesh sieve, wherein the drying temperature is 60 ℃.
(2) And (3) immersing the vinegar residue obtained in the step (1) into 2% dilute acetic acid solution for acidolysis, wherein the mass ratio of the vinegar residue to the dilute acetic acid is 1:20, the acidolysis temperature is 90 ℃, and the acidolysis time is 120min. Filtering, and washing the acidolysis vinegar residue with deionized water to neutrality.
(3) Adding a citric acid-sodium citrate buffer solution into the vinegar residue obtained in the step (2), wherein the mass ratio of the vinegar residue to the citric acid-sodium citrate buffer solution is 1:25, and regulating the pH value to be 4.8 to obtain a solution to be subjected to enzymolysis; adding cellulase for enzymolysis, wherein the mass ratio of the cellulase to the liquid to be subjected to enzymolysis is 1:200, the enzymolysis temperature is 50 ℃, the enzymolysis time is 6 hours, and filtering is carried out.
(4) Mixing the acidolysis solution and the enzymolysis solution obtained in the step (2) and the step (3), adding powdered activated carbon for detoxification and decoloration, wherein the mass ratio of the activated carbon to the mixed solution is 1:50, the temperature is 80 ℃, the stirring time is 40min, filtering and centrifuging.
(5) And (3) regulating the pH value of the mixed solution obtained in the step (4) to 5.0. Sterilizing at 121deg.C for 20min to obtain fermentation medium.
(6) And (3) inoculating Kang Pucha bacteria into the culture medium obtained in the step (5), and fermenting and culturing at 30 ℃ for 8 days, wherein the inoculum size is 6% of the volume of the culture medium.
(7) Purifying the bacterial cellulose obtained in the step (6) in a sodium hydroxide solution, wherein the mass ratio of the sodium hydroxide to the aqueous solution is 1:20, the temperature is 80 ℃, the time is 30min, and the purified bacterial cellulose membrane is obtained, wherein an SEM (scanning electron microscope) graph is shown in figure 1, a diameter distribution is shown in figure 2, an FTIR (light-emitting diode) graph is shown in figure 3, and an XRD (X-ray diffraction) graph is shown in figure 4.
Comparative example: the Kang Pucha bacteria were inoculated into a standard medium, hensman medium, and cultured at 30℃for 8 days with an inoculum size of 6% of the medium volume. The purification process is the same as above.
The purified bacterial cellulose prepared in both media was freeze-dried and dry weight was measured. Yield versus, for example, FIG. 5, it can be seen that bacterial cellulose production from vinegar residue medium will be approximately 5 times higher than standard medium production.
Example 2 preparation of bacterial cellulose Using Vinegar residues
The method comprises the following steps:
(1) The vinegar residue is washed and decontaminated, and is subjected to forced air drying, and the crushed vinegar residue is sieved by a 50-mesh sieve, wherein the drying temperature is 70 ℃.
(2) And (3) immersing the vinegar residue obtained in the step (1) into a dilute acetic acid solution with the concentration of 3% for acidolysis, wherein the mass ratio of the vinegar residue to the dilute acetic acid is 1:15, the acidolysis temperature is 100 ℃, and the acidolysis time is 100min. Filtering, and washing the acidolysis vinegar residue with deionized water to neutrality.
(3) Adding a citric acid-sodium citrate buffer solution into the vinegar residue obtained in the step (2), wherein the mass ratio of the vinegar residue to the citric acid-sodium citrate buffer solution is 1:20, and regulating the pH value to be 4.9 to obtain a solution to be subjected to enzymolysis; adding cellulase for enzymolysis, wherein the mass ratio of the cellulase to the liquid to be subjected to enzymolysis is 1:150, the enzymolysis temperature is 50 ℃, the enzymolysis time is 12 hours, and filtering is carried out.
(4) Mixing the acidolysis solution and the enzymolysis solution obtained in the step (2) and the step (3), adding powdered activated carbon for detoxification and decoloration, wherein the mass ratio of the activated carbon to the mixed solution is 1:30, the temperature is 90 ℃, the stirring time is 50min, filtering and centrifuging.
(5) And (3) regulating the pH of the mixed solution obtained in the step (4) to 6.0. Sterilizing at 121deg.C for 20min to obtain fermentation medium.
(6) And (3) inoculating Kang Pucha bacteria into the culture medium obtained in the step (5), and fermenting and culturing at 30 ℃ for 10 days, wherein the inoculum size is 8% of the volume of the culture medium.
(7) Purifying the bacterial cellulose obtained in the step (6) in a sodium hydroxide solution, wherein the mass ratio of the sodium hydroxide to the aqueous solution is 1:25, the temperature is 90 ℃, and the time is 40min, so as to obtain the purified bacterial cellulose membrane.
Example 3 preparation of bacterial cellulose Using Vinegar residues
The method comprises the following steps:
(1) Washing the vinegar residue, removing impurities, naturally drying, pulverizing, sieving with 60 mesh sieve, and drying at 70deg.C.
(2) And (3) immersing the vinegar residue obtained in the step (1) into 4% dilute acetic acid solution for acidolysis, wherein the mass ratio of the vinegar residue to the dilute acetic acid is 1:15, the acidolysis temperature is 110 ℃, and the acidolysis time is 90min. Filtering, and washing the acidolysis vinegar residue with deionized water to neutrality.
(3) Adding a citric acid-sodium citrate buffer solution into the vinegar residue obtained in the step (2), wherein the mass ratio of the vinegar residue to the citric acid-sodium citrate buffer solution is 1:15, and regulating the pH value to be 4.9 to obtain a solution to be subjected to enzymolysis; adding cellulase for enzymolysis, wherein the mass ratio of the cellulase to the liquid to be subjected to enzymolysis is 1:150, the enzymolysis temperature is 50 ℃, the enzymolysis time is 18 hours, and filtering is carried out.
(4) Mixing the acidolysis solution and the enzymolysis solution obtained in the step (2) and the step (3), adding powdered activated carbon for detoxification and decoloration, wherein the mass ratio of the activated carbon to the mixed solution is 1:25, the temperature is 90 ℃, the stirring time is 80min, filtering and centrifuging.
(5) And (3) taking the mixed solution obtained in the step (4) as a carbon source, and adjusting the pH to 5.0. Sterilizing at 121deg.C for 20min to obtain fermentation medium.
(6) Inoculating Kang Pucha bacteria into the culture medium obtained in the step (5), fermenting and culturing at 30 ℃ for 12 days, wherein the inoculum size is 10% of the volume of the culture medium.
(7) Purifying the bacterial cellulose obtained in the step (6) in a sodium hydroxide solution, wherein the mass ratio of the sodium hydroxide to the aqueous solution is 1:30, the temperature is 90 ℃, and the time is 50min, so as to obtain the purified bacterial cellulose membrane.
Example 4 preparation of bacterial cellulose Using Vinegar residues
The method comprises the following steps:
(1) Washing the vinegar residue, removing impurities, naturally drying, pulverizing, sieving with 80 mesh sieve, and drying at 80deg.C.
(2) And (3) immersing the vinegar residue obtained in the step (1) into a dilute acetic acid solution with the concentration of 5% for acidolysis, wherein the mass ratio of the vinegar residue to the dilute acetic acid is 1:10, the acidolysis temperature is 120 ℃, and the acidolysis time is 80min. Filtering, and washing the acidolysis vinegar residue with deionized water to neutrality.
(3) Adding a citric acid-sodium citrate buffer solution into the vinegar residue obtained in the step (2), wherein the mass ratio of the vinegar residue to the citric acid-sodium citrate buffer solution is 1:10, and regulating the pH value to be 5.0 to obtain a solution to be subjected to enzymolysis; adding cellulase for enzymolysis, wherein the mass ratio of the cellulase to the liquid to be subjected to enzymolysis is 1:100, the enzymolysis temperature is 50 ℃, the enzymolysis time is 24 hours, and filtering is carried out.
(4) Mixing the acidolysis solution and the enzymolysis solution obtained in the step (2) and the step (3), adding powdered activated carbon for detoxification and decoloration, wherein the mass ratio of the activated carbon to the mixed solution is 1:20, the temperature is 100 ℃, the stirring time is 100min, filtering and centrifuging.
(5) And (3) taking the mixed solution obtained in the step (4) as a carbon source, and adjusting the pH to 6.0. Sterilizing at 121deg.C for 20min to obtain fermentation medium.
(6) Inoculating Kang Pucha bacteria into the culture medium obtained in the step (5), fermenting and culturing at 30 ℃ for 14 days, wherein the inoculum size is 12% of the volume of the culture medium.
(7) Purifying the bacterial cellulose obtained in the step (6) in a sodium hydroxide solution, wherein the mass ratio of the sodium hydroxide to the aqueous solution is 1:25, the temperature is 100 ℃, and the time is 45min, so as to obtain the purified bacterial cellulose membrane.
The foregoing examples are provided to fully disclose and describe how to make and use the claimed embodiments by those skilled in the art, and are not intended to limit the scope of the disclosure herein. Modifications that are obvious to a person skilled in the art will be within the scope of the appended claims.
Claims (10)
1. A method for preparing bacterial cellulose by using vinegar residues, which is characterized by comprising the following steps:
(1) Taking vinegar residues, drying and crushing;
(2) Placing the dried and crushed vinegar residues into an acetic acid solution, and acidolyzing for 30-90 minutes at the temperature of 100-180 ℃; filtering to obtain acidolysis solution and acidolysis vinegar residue, and washing acidolysis vinegar residue with water to neutrality;
(3) Placing the acidolysis vinegar residues in water or buffer solution, and regulating the pH to 4.8-5.0 to obtain to-be-enzymatically hydrolyzed solution; adding cellulase, and carrying out enzymolysis for 4-24 hours at 48-52 ℃; filtering to obtain enzymolysis liquid;
(4) Mixing the acidolysis solution and the enzymolysis solution to obtain a mixed solution, and adding activated carbon to detoxify and decolor; filtering and centrifuging to obtain a fermentation medium;
(5) Sterilizing the fermentation medium, inoculating Kang Pucha bacteria, standing and fermenting at normal temperature for 7-14 days to obtain the bacterial cellulose membrane.
2. The method for preparing bacterial cellulose by using vinegar residues according to claim 1, wherein: in the step (1), the drying mode is selected from a natural drying method and a forced air drying method; the drying temperature is 60-80 ℃.
3. The method for preparing bacterial cellulose by using vinegar residues according to claim 1, wherein: in the step (2), the mass ratio of the vinegar residue to the acetic acid is 1:10-20.
4. The method for preparing bacterial cellulose by using vinegar residues according to claim 1, wherein: in the step (2), the concentration of the acetic acid solution is 1% -5%.
5. The method for preparing bacterial cellulose by using vinegar residues according to claim 1, wherein: in the step (3), the buffer solution is a citric acid-sodium citrate buffer solution, and the mass ratio of the vinegar residue to water or the buffer solution is 1:10-25.
6. The method for preparing bacterial cellulose by using vinegar residues according to claim 1, wherein: in the step (3), the mass ratio of the cellulase to the liquid to be subjected to enzymolysis is 1:50-200.
7. The method for preparing bacterial cellulose by using vinegar residues according to claim 1, wherein: in the step (4), the mass ratio of the activated carbon to the mixed solution is 1:20-100.
8. The method for preparing bacterial cellulose by using vinegar residues according to claim 1, wherein: in the step (5), the inoculation amount is 8% -15%.
9. The method for producing bacterial cellulose using vinegar residues according to any one of claims 1 to 8, further comprising the steps of: (6) Purifying bacterial cellulose in sodium hydroxide solution to obtain the purified bacterial cellulose membrane.
10. The method for preparing bacterial cellulose by using vinegar residues as claimed in claim 9, wherein: in the step (6), the concentration of the sodium hydroxide solution is 1-5%, the purification temperature is 80-100 ℃ and the time is 30-60 minutes.
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