CN110699414A - Method for preparing bacterial cellulose membrane by using enzymatic soybean hydrolysate - Google Patents
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- C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
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Abstract
A method for preparing a bacterial cellulose membrane by using enzymatic soybean hydrolysate, belonging to the technical field of cellulose membrane preparation. In order to solve the problem that the residual waste liquid of the soybean oil prepared by the enzymatic method cannot be reasonably utilized, the invention provides a method for preparing a bacterial cellulose membrane by utilizing enzymatic soybean hydrolysate, which comprises the following steps of (1) preparing an enzymatic soybean hydrolysate culture medium; (2) preparing a crude product of the bacterial cellulose membrane; (3) and (4) purifying the bacterial cellulose membrane. The method reasonably utilizes the residual waste liquid generated in the preparation of the soybean oil by the enzymatic method, does not need to treat a culture medium with acid water, and has the advantages of higher bacterial cellulose content, finer microfibril and higher maximum thermal degradation temperature, wherein the bacterial cellulose is synthesized by directly utilizing enzymatic soybean hydrolysate by a strain.
Description
Technical Field
The invention belongs to the technical field of bacterial cellulose membrane preparation, and particularly relates to a method for preparing a bacterial cellulose membrane by using enzymatic soybean hydrolysate.
Background
Bacterial Cellulose (BC) is a generic term for cellulose synthesized by any of microorganisms belonging to the genera Acetobacter (Acetobacter), Agrobacterium (Agrobacterium), Rhizobium (Rhizobium), Sarcina (Sarcina), and the like under different conditions. The bacterial cellulose forms a unique textile structure, and has the characteristics of high water absorption, high water retention, high transmittance to liquid and gas, high wet strength, in-situ processing and forming in particular in a wet state and the like due to the nanometer effect. The high purity and excellent performance enable the bacterial cellulose fiber to be widely applied in special fields.
At present, various industrial and agricultural wastes are studied as culture media for preparing bacterial cellulose, such as soybean molasses produced by processing concentrated protein, molasses produced by processing cane sugar or beet, and because bacterial strains for synthesizing bacterial cellulose utilize single glucose in the synthesis process of cellulose, high-strength acid hydrolysis (hydrochloric acid, nitric acid, sulfuric acid and the like) is required to be carried out on the molasses before the molasses is used as the culture media, so that the fermentation rate of the bacterial strains is improved. In addition, whey (albumin and albumin) produced by the production of soy protein isolate is also used, which is not beneficial to the utilization of the strain.
The enzymatic extraction of vegetable oil is a new oil extraction technology developed in the 70 s of the 20 th century, and as a new vegetable oil extraction technology, oil tissues and complexes such as lipoprotein, lipopolysaccharide and the like are subjected to enzymolysis on the basis of mechanical crushing, so that oil is liberated. The residual water phase of the enzymatic extraction of the vegetable oil still contains a plurality of valuable components, and the direct discarding avoids the waste of resources, so that the utilization of the residual water phase can be attempted.
Disclosure of Invention
The invention provides a method for preparing a bacterial cellulose membrane by using enzymatic soybean hydrolysate, aiming at solving the problem that the residual waste liquid in the enzymatic preparation of soybean oil cannot be reasonably utilized. The method comprises the following specific steps:
(1) preparation of an enzymatic soybean hydrolysate culture medium: taking the residual waste liquid of the soybean oil prepared by the enzyme method as a stock solution, adjusting the pH to 4.3-4.6, and removing impurities to be used as a culture medium;
(2) preparation of crude bacterial cellulose membrane: inoculating a fermentation strain into the liquid culture medium of the enzymatic soybean hydrolysate obtained in the step (1) for fermentation at the fermentation temperature of 25-32 ℃ for 8-15 days to obtain a bacterial cellulose membrane crude product;
(3) purification of bacterial cellulose membranes: boiling the crude bacterial cellulose membrane obtained in the step (2) with boiling water for 10-30min, boiling with 0.1-1M sodium hydroxide for 20-60min, soaking in water to adjust the pH value to be neutral, and changing water to wash until the cellulose membrane is transparent.
Preferably, the waste liquid in the step (1) is prepared by the following method: crushing soybean, tabletting, adding water to regulate quality to obtain soybean flour mucilage, extruding and puffing, mixing the obtained soybean flour with water to obtain soybean flour liquid, carrying out enzymolysis on the soybean flour liquid by using complex enzyme, and carrying out three-phase separation on the enzymolysis liquid to obtain hydrolysate.
Preferably, the step (1) of removing impurities is to filter the stock solution by a 300-400-mesh filter screen to remove impurities.
Preferably, the fermentation strain in the step (2) is black tea fungus.
Preferably, the concentration of the fermentation strain inoculated in the step (2) is 6X 106-6×108cfu/ml。
Preferably, the temperature conditions of the extrusion and expansion are that the first section is 40-60 ℃, the second section is 60-80 ℃, the third section is 80-95 ℃ and the fourth section is 95-105 ℃.
Preferably, the temperature conditions of the extrusion and expansion are that the first section is 48 ℃, the second section is 72 ℃, the third section is 86 ℃ and the fourth section is 102 ℃.
Preferably, the compound enzyme enzymolysis refers to enzymolysis by using a mixed enzyme soybean powder solution of alkaline protease accounting for 4-8 per mill of the mass fraction of the soybean powder and flavourzyme accounting for 1-4 per mill of the mass fraction of the soybean powder.
Preferably, the compound enzyme enzymolysis is to carry out enzymolysis on the soybean powder liquid by using a mixed enzyme of alkaline protease accounting for 5 per mill of the mass fraction of the soybean powder and flavourzyme accounting for 2 per mill of the mass fraction of the soybean powder.
Preferably, the three-phase separation is performed by using a three-phase horizontal separator under the conditions that the centrifugal rotating speed is 4300-.
Advantageous effects
The method reasonably utilizes the residual waste liquid generated in the preparation of the soybean oil by the enzymatic method, does not need to treat a culture medium with acid water, and has the advantages of higher bacterial cellulose content, finer microfibril and higher maximum thermal degradation temperature, wherein the bacterial cellulose is synthesized by directly utilizing enzymatic soybean hydrolysate by a strain.
Drawings
FIG. 1 shows the results of three-phase horizontal separation, wherein a is the separation result of example 1, b is the separation result of example 2, and c is the separation result of example 3;
FIG. 2 is a scanning electron microscope observation result of the bacterial cellulose obtained in example 1;
FIG. 3 is a scanning electron microscope observation result of the bacterial cellulose obtained in example 4;
FIG. 4 statistical results of diameters of bacterial cellulose obtained in example 1;
FIG. 5 statistical results of diameters of bacterial cellulose obtained in example 4;
FIG. 6 is an atomic force microscope observation of the bacterial cellulose obtained in example 1;
FIG. 7 is an atomic force microscope observation of the bacterial cellulose obtained in example 4;
FIG. 8 shows the thermal degradation temperature results of the bacterial cellulose obtained in example 1;
FIG. 9 shows the thermal degradation temperature results of the bacterial cellulose obtained in example 4.
Detailed Description
The present invention will be further described with reference to the following specific examples, but the present invention is not limited to these examples.
The materials, reagents, methods and apparatus used in the following examples, which are not specifically illustrated, are conventional in the art and are commercially available to those skilled in the art. The black tea fungus is produced by black tea fungus biological cultivation limited company of the beneficial fungus old and well-known family.
Example 1. a method for preparing a bacterial cellulose membrane using an enzymatic soy hydrolysate.
The method for preparing the bacterial cellulose membrane by using the enzymatic soybean hydrolysate comprises the following specific steps:
(1) preparation of an enzymatic soybean hydrolysate culture medium:
the enzymatic soybean hydrolysate is waste liquid left by preparing soybean oil by an enzymatic method, is hydrolysate obtained by crushing, tabletting, tempering, extruding and puffing, crushing and sieving, carrying out compound enzymolysis and three-phase separation on soybeans, and specifically comprises the following steps:
1) crushing soybean into 5 pieces;
2) pressing into bean slices with thickness of 3 mm;
3) mixing the bean slices obtained in the step 2) with water, wherein the mass fraction of the water is 10.5%, and obtaining soybean meal mucilage;
4) extruding and puffing the soybean meal mucilage obtained in the step 3), wherein the puffing temperature is 48 ℃ in the first section, 72 ℃ in the second section, 86 ℃ in the third section and 102 ℃ in the fourth section;
5) crushing the extruded and puffed soybean flour obtained in the step 4), and sieving the crushed soybean flour with a 70-mesh sieve, wherein the sieved soybean flour is mixed with water according to the weight ratio of 1 g: mixing 6.5ml of the mixture to prepare soybean powder liquid;
6) mixing alkaline protease 5 ‰ of semen glycines powder and flavourzyme 2 ‰ of semen glycines powder with the prepared semen glycines powder solution, hydrolyzing at 55 deg.C for 5 hr, and inactivating enzyme at 94 deg.C for 3 min.
7) Adopting three horizontal separators, 4400rad/min, centrifuging for 16s to obtain hydrolysate, and obtaining a three-phase horizontal separation result as shown in a in figure 1, wherein the lowest layer is the soybean hydrolysate by the enzyme method, sterilizing the hydrolysate at 121 ℃ for 19min, and filtering the sterilized hydrolysate by a 300-mesh filter screen to remove precipitated impurities to obtain the soybean hydrolysate by the enzyme method.
Adjusting the pH value of the enzymatic soybean hydrolysate obtained in the process to 4.5, filtering by using a 300-mesh filter screen to remove impurities, and sterilizing the filtrate to obtain the enzymatic soybean hydrolysate liquid culture medium.
(2) Preparation of crude bacterial cellulose membrane:
1) preparing a standard HS culture medium, wherein the components of the culture medium are glucose (20g/L), yeast extract powder (5g/L), peptone (5g/L), citric acid (1.15g/L) and disodium hydrogen phosphate (2.7g/L), and the pH value is 6.0;
2) inoculating 6% black tea fungus by volume fraction into the HS culture medium in the step 1), culturing at 28 ℃ for 24h, and completing activation and rejuvenation of the strain;
3) preparing a seed solution, wherein the components of the seed solution are that the mass concentration of glucose is 40g/L, and the mass concentration of black tea is 5 g/L;
4) inoculating the black tea fungus activated in the step 2) into seed liquid, wherein the inoculation volume is 6% of the volume of the seed liquid, culturing for 24h at 28 ℃, and culturing until the concentration of the strain is 107cfu/ml。
Inoculating the strain into the liquid culture medium of the enzymatic soybean hydrolysate obtained in the step (1) for fermentation, wherein the inoculation volume fraction is 6%, the fermentation temperature is 28 ℃, and the fermentation time is 13 days, so as to obtain a crude bacterial cellulose membrane.
(3) Purification of bacterial cellulose membranes:
boiling the cellulose membrane obtained in the step (2) with water for 20min, then boiling with 0.2M sodium hydroxide for 40min, soaking in water to adjust the pH value to be neutral, and changing water to wash until the cellulose membrane is transparent.
Example 2. a method of preparing a bacterial cellulose membrane using an enzymatic soy hydrolysate.
The method for preparing the bacterial cellulose membrane by using the enzymatic soybean hydrolysate comprises the following specific steps:
(1) preparation of an enzymatic soybean hydrolysate culture medium:
the enzymatic soybean hydrolysate is waste liquid left by preparing soybean oil by an enzymatic method, is hydrolysate obtained by crushing, tabletting, tempering, extruding and puffing, crushing and sieving, carrying out compound enzymolysis and three-phase separation on soybeans, and specifically comprises the following steps:
1) crushing the soybeans into 4 pieces;
2) pressing into bean slices with thickness of 2 mm;
3) mixing the bean slices obtained in the step 2) with water, wherein the water accounts for 10 mass percent, and obtaining soybean meal mucilage;
4) extruding and puffing the soybean meal mucilage obtained in the step 3), wherein the puffing temperature is 40 ℃ in the first section, 60 ℃ in the second section, 80 ℃ in the third section and 95 ℃ in the fourth section;
5) crushing the extruded and puffed soybean flour obtained in the step 4), and sieving the crushed soybean flour with a 60-mesh sieve, wherein the sieved soybean flour is mixed with water according to the weight ratio of 1 g: mixing 6ml of the above materials to obtain soybean powder;
6) mixing alkaline protease 4 ‰ of semen glycines powder and flavourzyme 1 ‰ of semen glycines powder with the prepared semen glycines powder solution, hydrolyzing at 50 deg.C for 2 hr, and inactivating enzyme at 90 deg.C for 3 min.
7) Adopting three horizontal separators, 4300rad/min, centrifuging for 10s to obtain hydrolysate, and obtaining a three-phase horizontal separation result as shown in b in figure 1, wherein the lowest layer is the soybean hydrolysate by the enzyme method, sterilizing the hydrolysate at 121 ℃ for 18min, and filtering the sterilized hydrolysate by a 300-mesh filter screen to remove precipitated impurities to obtain the soybean hydrolysate by the enzyme method.
Adjusting the pH value of the enzymatic soybean hydrolysate obtained in the process to 4.3, filtering by using a 300-mesh filter screen to remove precipitated impurities, and sterilizing the filtrate to obtain the enzymatic soybean hydrolysate liquid culture medium.
(2) Preparation of crude bacterial cellulose membrane:
1) preparing a standard HS culture medium, wherein the components of the culture medium are glucose (20g/L), yeast extract powder (5g/L), peptone (5g/L), citric acid (1.15g/L) and disodium hydrogen phosphate (2.7g/L), and the pH value is 6.0;
2) inoculating 6% black tea fungus by volume fraction into the HS culture medium in the step 1), culturing at 25 ℃ for 20h, and completing activation and rejuvenation of the strain;
3) preparing a seed solution, wherein the components of the seed solution are that the mass concentration of glucose is 20g/L, and the mass concentration of black tea is 3 g/L;
4) inoculating the activated black tea fungus obtained in the step 2) into a seed solution, wherein the inoculation volume is 6% of the volume of the seed solution, culturing at 25 ℃ for 20h until the concentration of the fungus strain is 106cfu/ml。
Inoculating the strain into the liquid culture medium of the enzymatic soybean hydrolysate obtained in the step (1) for fermentation, wherein the inoculation volume fraction is 6%, the fermentation temperature is 25 ℃, and the fermentation time is 8 days, so as to obtain a crude bacterial cellulose membrane.
(3) Purification of bacterial cellulose membranes:
boiling the cellulose membrane obtained in the step (4) with water for 10min, then boiling with 0.1M sodium hydroxide for 20min, soaking in water to adjust the pH value to be neutral, and changing water to wash until the cellulose membrane is transparent.
Example 3. a method of preparing a bacterial cellulose membrane using an enzymatic soy hydrolysate.
The method for preparing the bacterial cellulose membrane by using the enzymatic soybean hydrolysate comprises the following specific steps:
(1) preparation of an enzymatic soybean hydrolysate culture medium:
the enzymatic soybean hydrolysate is waste liquid left by preparing soybean oil by an enzymatic method, is hydrolysate obtained by crushing, tabletting, tempering, extruding and puffing, crushing and sieving, carrying out compound enzymolysis and three-phase separation on soybeans, and specifically comprises the following steps:
1) crushing soybean into 6 pieces;
2) pressing into bean slices with thickness of 4 mm;
3) mixing the bean slices obtained in the step 2) with water, wherein the water accounts for 11% by mass, and obtaining soybean meal mucilage;
4) extruding and puffing the soybean meal mucilage obtained in the step 3), wherein the puffing temperature is 60 ℃ in the first section, 80 ℃ in the second section, 95 ℃ in the third section and 105 ℃ in the fourth section;
5) crushing the extruded and puffed soybean flour obtained in the step 4), and sieving the crushed soybean flour with a 80-mesh sieve, wherein the sieved soybean flour is mixed with water according to the weight ratio of 1 g: mixing 7ml of the above materials to obtain soybean powder;
6) mixing alkaline protease 8 ‰ of semen glycines powder and flavourzyme 4 ‰ of semen glycines powder with the prepared semen glycines powder solution, hydrolyzing at 60 deg.C for 6 hr, and inactivating enzyme at 90 deg.C for 3 min.
7) And (3) centrifuging at 4500rad/min for 20s by using a three-phase horizontal separator to obtain a hydrolysate, wherein the three-phase horizontal separation result is shown as c in figure 1, the lowest layer is the enzymatic soybean hydrolysate, sterilizing the hydrolysate at 121 ℃ for 20min, and filtering the sterilized hydrolysate by using a 400-mesh filter screen to remove precipitated impurities to obtain the enzymatic soybean hydrolysate.
Adjusting the pH value of the enzymatic soybean hydrolysate obtained in the process to 4.6, filtering by adopting a 400-mesh filter screen to remove precipitated impurities, and sterilizing the filtrate to obtain the enzymatic soybean hydrolysate liquid culture medium.
(2) Preparation of crude bacterial cellulose membrane:
1) preparing a standard HS culture medium, wherein the components of the culture medium are glucose (20g/L), yeast extract powder (5g/L), peptone (5g/L), citric acid (1.15g/L) and disodium hydrogen phosphate (2.7g/L), and the pH value is 6.0;
2) inoculating 6% black tea fungus by volume fraction into the HS culture medium in the step 1), culturing at 32 ℃ for 48h, and completing activation and rejuvenation of the strain;
3) preparing a seed solution, wherein the components of the seed solution are that the mass concentration of glucose is 60g/L, and the mass concentration of black tea is 7 g/L;
4) inoculating the black tea fungus activated in the step 2) into seed liquid, wherein the inoculation volume is 6% of the volume of the seed liquid, culturing for 48h at 32 ℃, and culturing until the concentration of the strain is 108cfu/ml。
Inoculating the strain into the liquid culture medium of the enzymatic soybean hydrolysate obtained in the step (1) for fermentation, wherein the inoculation volume fraction is 6%, the fermentation temperature is 32 ℃, and the fermentation time is 15 days, so as to obtain a crude bacterial cellulose membrane.
(3) Purification of bacterial cellulose membranes:
boiling the cellulose membrane obtained in the step (4) with boiling water for 30min, then boiling with 1M sodium hydroxide for 60min, soaking in water to adjust the pH value to be neutral, and changing water to wash until the cellulose membrane is transparent.
Example 4 comparative example to example 1.
The same as example 1, except that in the crude preparation of the bacterial cellulose membrane in step (2) of this example, the fermentation medium used in 4) was also a standard HS medium, and a bacterial cellulose membrane was prepared.
Compared with the bacterial cellulose synthesized in the embodiment 1 and the embodiment 4, the yield of the cellulose prepared by the culture medium of the enzymatic soybean hydrolysate in the embodiment 1 is 1.78g/L, and is improved by 30.0 percent compared with the yield of the standard HS culture medium in the embodiment 4 which is 1.25 g/L;
the surface morphology of the freeze-dried bacterial cellulose was observed by an SEM scanning electron microscope (SEM, SU8010, HITACHI), and the results showed that the bacterial cellulose consisted of rod-shaped nanofibers and formed a porous three-dimensional network structure. The results obtained by scanning electron microscopy in example 1 are shown in FIG. 2 and the results obtained by scanning electron microscopy in example 4 are shown in FIG. 3, and comparing the two figures, it is seen that the fibers produced by the standard HS medium of example 4 are slightly coarser, while the bacterial fibers produced by the enzymatic soy hydrolysate medium of example 1 are slightly finer. The type of surface medium affects the morphological properties of the fibers in the bacterial cellulose.
The diameters of the cellulose obtained in example 1 and example 4 were counted, the statistical result of example 1 is shown in fig. 4, the statistical result of example 4 is shown in fig. 5, the fiber diameters of the cellulose in fig. 4 and fig. 5 are distributed between 40nm and 200nm, the fiber diameter obtained by the medium of enzymatic soybean hydrolysate in example 1 is 100nm, the average fiber diameter of the standard HS medium in example 4 is 114nm, and the microfiber of the bacterial cellulose obtained by the medium of enzymatic soybean hydrolysate in example 1 is finer compared with the fiber diameter of the bacterial cellulose.
The atomic force microscope can be used for observing the microscopic morphology and microscopic details of the surface of the bacterial cellulose, the compact and aggregated typical structure of the freeze-dried bacterial cellulose can be observed more clearly, the atomic force microscope shows a nanoscale network structure, and the bacterial cellulose microfibers are closely stacked and arranged irregularly. The three-dimensional structure of the cellulose obtained in example 1 was observed by an AFM atomic force microscope (Bruker, Germany) and the results are shown in FIG. 6 and that of the cellulose obtained in example 4 and that of the bacterial cellulose produced in the medium for comparison with the standard HS medium and the medium for enzymatic hydrolysis of soybean in example 1 are shown in FIG. 7, in which the width of the diameter of the microfibril was different and the diameter of the microfibril formed in the standard HS medium was wider and was consistent with the results observed by an SEM electron microscope.
The maximum thermal degradation temperature was analyzed by TGA thermogravimetric analyzer (Pyris 6 TGA, Perkin Elmer co., ltd. The results of example 1 are shown in fig. 8 and the results of example 4 are shown in fig. 9, the bacterial cellulose exhibits two distinct thermal degradation stages, the first of which occurs at 90 ℃ to 100 ℃, mainly the amount of absorbed moisture on the surface and the loss of interlayer coordinated water molecules. The second thermal degradation stage occurs at 300 ℃ to 400 ℃, which is thermal degradation and cracking of the bacterial cellulose framework, and finally, the bacterial cellulose is decomposed into water, carbon dioxide and the like. Example 4 the thermal degradation rate temperatures of the bacterial cellulose produced in the standard HS medium and the enzymatic soy hydrolysate medium of example 1 were 331.67 deg.C and 338.89 deg.C, respectively. The thermal degradation rate temperature increased by 7.22 ℃.
The cellulose yield, morphology, diameter and degree of thermal degradation obtained for example 2 and example 3 were similar to those of example 1.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (10)
1. A method for preparing a bacterial cellulose membrane by using enzymatic soybean hydrolysate is characterized by comprising the following steps:
(1) preparation of an enzymatic soybean hydrolysate culture medium: taking the residual waste liquid of the soybean oil prepared by the enzyme method as a stock solution, adjusting the pH to 4.3-4.6, and removing impurities to be used as a culture medium;
(2) preparation of crude bacterial cellulose membrane: inoculating a fermentation strain into the liquid culture medium of the enzymatic soybean hydrolysate obtained in the step (1) for fermentation at the fermentation temperature of 25-32 ℃ for 8-15 days to obtain a bacterial cellulose membrane crude product;
(3) purification of bacterial cellulose membranes: boiling the crude bacterial cellulose membrane obtained in the step (2) with boiling water for 10-30min, boiling with 0.1-1M sodium hydroxide for 20-60min, soaking in water to adjust the pH value to be neutral, and changing water to wash until the cellulose membrane is transparent.
2. The method for preparing the bacterial cellulose membrane by using the enzymatic soybean hydrolysate according to claim 1, wherein the waste liquor in the step (1) is prepared by the following method: crushing soybean, tabletting, adding water to regulate quality to obtain soybean flour mucilage, extruding and puffing, mixing the obtained soybean flour with water to obtain soybean flour liquid, carrying out enzymolysis on the soybean flour liquid by using complex enzyme, and carrying out three-phase separation on the enzymolysis liquid to obtain hydrolysate.
3. The method for preparing bacterial cellulose membrane by using enzymatic soybean hydrolysate as claimed in claim 1, wherein the step (1) of removing impurities comprises filtering the stock solution by using a 300-mesh 400-mesh filter screen.
4. The method for preparing a bacterial cellulose membrane by using the enzymatic soybean hydrolysate according to claim 1, wherein the fermentation strain of the step (2) is black tea fungus.
5. The method for preparing bacterial cellulose membrane by using enzymatic soybean hydrolysate according to claim 1, wherein the concentration of inoculated fermentation strain in the step (2) is 6 x 106-6×108cfu/ml。
6. The method for preparing a bacterial cellulose membrane from the enzymatic soybean hydrolysate according to claim 2, wherein the temperature conditions for the extrusion and expansion are 40-60 ℃ in one section, 60-80 ℃ in the second section, 80-95 ℃ in the third section and 95-105 ℃ in the fourth section.
7. The method for preparing a bacterial cellulose membrane by using the enzymatic soybean hydrolysate as claimed in claim 6, wherein the temperature conditions of the extrusion and expansion are 48 ℃ in one section, 72 ℃ in the second section, 86 ℃ in the third section and 102 ℃ in the fourth section.
8. The method for preparing the bacterial cellulose membrane by using the enzymatic soybean hydrolysate as claimed in claim 2, wherein the compound enzyme enzymolysis is carried out by using a mixed enzyme soybean powder solution of alkaline protease accounting for 4-8% of the soybean powder by mass and flavourzyme accounting for 1-4% of the soybean powder by mass.
9. The method for preparing the bacterial cellulose membrane by using the enzymatic soybean hydrolysate as claimed in claim 8, wherein the complex enzyme enzymolysis is carried out on the soybean powder liquid by using a mixed enzyme of alkaline protease accounting for 5 per mill of the mass fraction of the soybean powder and flavourzyme accounting for 2 per mill of the mass fraction of the soybean powder.
10. The method for preparing bacterial cellulose membrane by using enzymatic soybean hydrolysate as claimed in claim 2, wherein the three-phase separation is performed by using a three-phase horizontal separator at 4300-.
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