WO2014133249A1 - Method for preparing microbial cellulose gel - Google Patents

Abstract

The present invention provides a method for preparing a microbial cellulose gel, which is characterized by introducing a microbial strain into a medium and then culturing the microbial strain to prepare a first cellulose gel which is first gelled, pulverizing the first cellulose gel to obtain micronized cellulose, and subjecting the micronized cellulose to second gelation to obtain a second cellulose gel. The cellulose gel prepared according to the present invention has a more micronized fiber length but similar moisture content or density as compared with the first cellulose gel which is first gelled, thereby manufacturing the same quality of cellulose product, and can decrease the amounts of raw materials used and remarkably increase the production yield, thereby significantly improving the competitiveness of microbial cellulose-related products.

Description

 Specification

 Title of invention: Microbial Cellulose Gel Manufacturing Method

 [1] The present invention relates to a method for producing a microbial salulose gel, specifically

 Micronized by pulverizing the salose gel produced first through microbial culture

 A method of producing a cellulose gel having excellent productivity and quality by obtaining cellulose and re-secondary gelling is proposed.

 [2]

 Background

 [3] Cellulose, the basic structural component of plant cell walls, contains 33% of all plant material.

 It is the most abundant organic compound produced in nature.

 Salolos is processed to produce paper and fibers, and because it can be chemically transformed into materials that make commodities such as plastics, photographic films and rayon

 It is an economically important substance. Other cellulose derivatives are used in adhesives, explosives, food thickeners and moisture proof coatings.

 [4] Cellulose is the most abundant biological material resource in nature, especially acetic acid bacteria.

 Since the production of salulose has been reported, bacterial cells produced by microorganisms (bacterial cellulose) has been the subject of constant research as a new material, microbial bacterium salulose is not only an additive in food but also ， It is attracting much attention as a new value-added industrial material.

 [5] The plant-derived salose and the bacterium sallose make a big structural difference. Microbial-derived bacterial cells

 With beta-di-glucopyranose (β-D-glucopyranose)

 Long-chains bound by beta-1,4-glucoside ((3-1,4-glucoside) are in the form of ribbon-like bundles in which hydrogen bonds are maintained, microfibrils

 It is structurally different from salose derived from plants in the form of bundles.

In addition, unlike plant-derived cellulose, bacterial salulose contains no ingredients other than salulose, and is easy to isolate and purify. In other words, the plant has a crystalline structure containing hemicellulose, lignin, etc., as well as salulose.In the case of bacterial cellulose, other impurities other than salulose produced by bacteria Since it is not included, it is easy to separate and purify it.

[7] Microbial-derived bacterial cellulose is the only natural salulose that can be obtained on film. Gel-like microbial salulose sheets are not only very similar to the film but also more dense and three-dimensional than pulp. As it has a network structure, it has high absorbency and water retention, and it is possible to develop a product utilizing this property. have. For example, microbial cellulose films produced by culturing acetobacter genus microorganisms were subjected to post-processing and pressing in a gel state to reduce moisture content. It can be used as a cosmetic mask pack or a medical burn treatment. In particular, the microbial salose of the gel state is easy to contain an additive therein, and is effective in expressing various functionalities.

 [8] The conventional film-type microbial salose sheet was manufactured by the following method. First, as shown in FIG. 1, a medium is prepared and inoculated with appropriate bacteria, followed by culturing for a predetermined period in a culture chamber in which temperature and humidity are controlled to obtain a cell-shaped bulk sheet formed in a gel state in an upper portion of the culture medium. .

 [9] The bulk microbial salloose sheet thus prepared was surfaced during culturing.

 There is a problem such as damage that does not grow clean and evenly occurs (see Fig. 2). In this case, since undesired poor microbial cells must be completely disposed of, the amount of cellulose sheets that can be used as a real product is inevitably deteriorated.

 [10] In addition, the bulk microbial cells must be processed to cut into a suitable thickness, for example, to be used as a mask pack, and to slice cells with low physical strength uniformly into thin thicknesses. Is a very difficult process. In particular, microorganisms grown in uneven thickness

 Cellulose accelerates the problem of yield degradation due to the small amount of sheets that can be used as actual products during the slicing process.

 [11] Finally, despite the various functions and value of microbial salulose, there are limitations in improving the yield of actual products due to the quality problems of the bulk microbial cells produced and the difficulty of the slicing process. Therefore, in order to develop an industry utilizing microbial cellloose, development of microbial cellloose production and processing technology is more urgently needed.

 [12]

 Detailed description of the invention

 Technical challenges

 The present invention has been made under the technical background described above, and an object of the present invention is to provide a new method for preparing microbial cell rose gel.

In addition, another object of the present invention is to provide a method for producing a secondary microbial cell-loose gel in which the function and effect are not degraded by utilizing a primary microorganism salulose gel that has surface damage during the culture process. will be.

In addition, another object of the present invention is to improve the production yield of microbial salulose gel to promote the expansion and development of the industry utilizing microbial cells.

[16] In addition, still another object and technical features of the present invention in the following detailed description It will be presented in detail.

 [17]

 Challenge solution

 In order to achieve the above object, the present invention is prepared by introducing a microorganism strain into the medium, followed by culturing to prepare a first gelled first cell cellulose gel, and by grinding the first cell cellulose gel to finely divided ) To obtain a micronized salose, and to micronize the micronized salose to prepare a second cell rose gel.

 [19] Preferably, the second cell cellulose gel is prepared at a temperature of 60 ° C. or higher, and the salose obtained by pulverizing the first cell cellulose gel and micronized is, for example.

 It can be gelled by adding an excipient. In the gelation process of finely divided salulose, a large amount of water may be further included, and other functional additives may be further included to change the functionality of the cellulose gel.

 [2] The second salose gel may be compressed to improve density and reduce moisture content. The compressed second cellulose gel may be sliced to a thin thickness and processed into a product for a mask pack or a medical therapeutic through a molding process.

 [21] The second cell rose gel has a higher density of fibers than the first cell rose gel.

 The water content remains the same while being lowered, and the density can be maintained to the same or similar degree.

 [22] The present invention also provides a salose gel prepared by the above-described method, in which the length of fiber in cellulose is finely divided into 0.1 to lOimn, and the water content of the fiber is 1:99. It provides a micronized fibrous salulose gel, characterized in that in the range of ~ 30:70.

 [23]

 Effects of the Invention

 [24] According to the present invention, the second finally obtained through two gelling processes

 While the salose gel has a finer fiber length than the first gelled first salose gel, the moisture content and density are similar to produce a cellulose product of the same quality.

[25] In addition, it is possible to contribute to cost reduction by reducing the amount of raw materials used in the production of microbial salulose gel-related products, and to significantly increase the production yield of salulose gels, thereby greatly improving the competitiveness of microbial cellulose-related products. Can be.

In addition, the cellulose gel has excellent moldability, which makes it easy to process the application of salulose, and may be applied to various fields by including functional additives and pigments.

 [27]

Brief description of the drawings 1 is a manufacturing process diagram of a conventional microbial cell rose gel

 [29] Figure 2 is a photograph showing the surface damage of the prepared bulk cell cellulose gel

 3 is a manufacturing process diagram of a microorganism salulose gel according to the present invention

 [31] Figure 4 is a photograph showing a first salose gel

 5A and 5B are photographs showing micronized salolos

 6A and 6B are surface and cross-sectional microstructure photographs of a first salose gel

 7A to 7C are microstructure photographs of a second cell rose gel according to grinding time.

 8 is a photograph showing a mask pack prepared from a second salose gel

 9 is a photograph showing a mask pack made of a first cellulose gel

 10 is a photograph showing another mask pack made of a second salose gel

 [38]

 Best mode for carrying out the invention

 [39] The present invention is the first gelled first by culturing the microorganism strain in the medium

 Preparing a cellose gel; Grinding the first salose gel in a state in which water is mixed to obtain finely divided cells out of the gel state; Water was mixed with the micronized cell out of the gel state, and a second gel was prepared by secondary gelling by adding an excipient and stirring at a temperature of 60 ° C. or higher; The excipient is a microorganism characterized in that it is a gelling agent selected from galactomannan, glucomannan, guar gum, locust bean gum, pluronic, baby, algin, carrageenan, xanthan gum, tara gum, tamarind gum, gellan A method of preparing a salose gel is proposed.

 [40]

 Form for the implementation of the invention

 [41] The present invention relates to a novel microbial salulose gel manufacturing process, and proposes a method for producing a cellulose gel, in which raw materials are reduced and mass production is easy while maintaining physical properties and quality through a second gelation process. do.

 Specifically, the present invention is to prepare a microbial cell with high crystallinity, excellent mechanical strength, adsorptivity, water retention, etc., the secondary cell after pulverizing the primary cellulose gel obtained through the political vessel ¾ Gelling increases the yield of cellulose gels while improving processability and product functionality.

 A process for preparing a microbial cell cellulose gel according to the present invention is as shown in FIG. 3.

 First, a cellulose culture process by microbial fermentation is performed (step S110). A culture medium is prepared by filling a prepared medium and a microorganism strain. Specifically, a medium for producing a bacterial cell in which one or more sucrose, glucose, and fructose supplements is added as a carbon source is prepared, and the medium is inoculated with the strain. After incubation in aerobic conditions.

[45] Microbial cell-producing strains include Acetobacter sp., Agrobacterium sp., Rhizobium sp. And Pseudomonas. The genus Pseudomonas sp. And Sarcina sp. Include the acetobacter xylium, acetobacter pasteurinus and aceto, especially among the genus Acetobacter sp. A. hansenii is known a lot. The present invention does not particularly limit the strain used to produce microbial salose, and various strains may be used depending on the type of culture medium.

 In addition, as an auxiliary carbon source for the production of microbial salulos, citrus and persimmon vinegar, apple juice, grape juice, citrus juice, beer waste liquid, coconut by-products, and the like may be used. It does not limit the type of natural materials used in the process. ᅳ

[47] A bacterial cell is obtained from the culture solution. As suitable culture conditions, the incubation temperature is maintained at a temperature in the range of 10 to 40 ° C., preferably 25 to 35 ° C., and the culture period is maintained at 5 to 20 days. In order to create aerobic conditions in the culture, the oxygen concentration in the culture chamber may be maintained in the range of l-100% (w / w), preferably 20-80% (w / w).

 The culture of the first cell to obtain a cellulose gel (step S120), the resulting first cellulose gel is a bulk of the thick film form, as shown in Figure 4 in a constant density and moisture content Keep it. Depending on the culture period, the first cellulose may have a thickness of about 5 mm to about 20 mm.

 [49] The first salose gel obtained was subjected to pulverization of undifferentiated cells through pulverization.

 (Step S130). Grinding | pulverization can grind | pulverize a bulk type salulose gel in the range of 0.1-10 mm of fiber length through an industrial mixer etc., for example. A large amount of water may be added in the process of micronization through milling, and water may be subsequently added after milling. The finely divided cellulose after grinding may have an increased water content and a lower density when compared with the first cellulose rose.

 [50] The appearance of micronized salose is shown in FIGS. 5A and 5B. The first salose gel used for grinding may be used as well as cells grown normally through culture, as well as the poorly grown cells shown in FIG. 2A. The pulverization method for the micronization of the first cell cellulose gel is not particularly limited, and may be chemically treated by physical crushing, pressing, stirring or the like, or by a mixed reaction with the additives.

 [1] Undifferentiated Cellulose is Secondarily Gelled to Prepare a Second Cellulose Gel (Step

 S140). The second salose gel can be gelated by adding an excipient, for example, to the cell finely divided by milling the first salose gel.

Excipients include, for example, galactomannan, glucomannan, guar gum, locust bean gum, pullulonic, agar, algin, carrageenan, xanthan gum, tara gum, tamarind gum, gellan, etc. Gelling agents may be used, especially excipients There is no limit to the kind.

 In the process of gelling micronized cells, a large amount of water may be further included, and other functional additives may be further included to change the functionality of the cellulose gel.

 The second cell cellulose gel is preferably prepared at a temperature of 60 ° C. or higher for facilitation and uniform mixing of the micronized cells with the substance added for releasing the micronized cells.

 [54] The second salose gel obtained has a higher density of fibers than the first cellulose gel.

 Lower, the moisture content remains the same, and the density remains the same or a similar degree. In addition, while the length of the fiber in the second salose gel is reduced through the micronization process, the water content compared to the fiber is maintained in the range of 1:99 ~ 30:70 due to the water added in the second gelling process.

 The manufactured second cell cellulose gel may be imparted with functionality and moldability to be utilized in various products through a post process (step S150). For example, the second salose gel may be compressed to improve density and reduce moisture content. In addition, the compressed second cellulose gel may be sliced to a thin thickness or processed into a product for a mask pack or a medical therapeutic product through a molding process. Through the pressing process, the physical strength such as the tensile strength of the second salose gel may be increased, and the moisture content of the second salose gel may be controlled through the water discharge. For example, in the case of the first cell cellulose gel, the water content is 98%, the fiber is about 2% ratio, the second cellulose gel may be pressed to adjust the water content at a rate of 50 to 60%.

 [56] In the subsequent process for such processing or gelling the second cell with a rose gel

 The process may be sterilized or further include functional additives such as emulsifiers (Beeswax, Tween # 60, etc.), preservatives (MP, PP, Saliethan, etc.), moisturizers (Glycerin, etc.).

 [57] In general, microbial salorose obtained through cultivation is difficult to process slices and the like, and yield of raw materials is low due to low yield of normal products, while there is a limit in mass production, but is obtained through grinding and secondary gelation. The microbial salulose gel of the invention greatly reduces the amount of raw materials to be introduced, which significantly increases the moldability for cost reduction and commercialization.

 [58]

 [59] Hereinafter, the features of the present invention in more detail through the preferred embodiment

 Explain.

 [60] Apocrypha Example 1-Undifferentiated Seminarmother Gem

As a strain for producing bacterial salolos, the KCG326 strain of the genus Gluconacetobacter was used. Glucose 10% (w / w), yeast extract l% (w / w), CaC0 ₃ 2% (w /), agar 1.5%> (w / w), remaining purified water Basal medium was prepared by mixing pH 6.8.

[62] SPL Incu for plant tissue culture containing 200 m £ of 272 medium as a preculture Inoculate one platinum strain from a flat plate agar medium into a tissue container

 Incubated at 27 ° C. for 48 hours. After inoculating the pre-culture solution l <¾ (w / w) in a tray containing the culture broth 1,300ι was incubated for 14 days at 30 ° C.

 [0063] The resulting bulk bacterial salose in gel was isolated from the culture. As shown in FIG. 4, the separated cellulose gel (the first cellulose gel) had a thick film form and a normal growth and abnormal growth (nonuniform growth).

 [64] The separated first cell was added to the blender gel in a blender for 5 to 10 minutes.

 It was crushed to obtain micronized salose. The obtained micronized cell rose out of the gel state and took the form of a mixed solution dispersed in water.

 [65] 75 wt% of distilled water (DI water) was added to the micronized salose, and used as an excipient.

 Carrageenan and xanthan gum were added to lwt% and 0.5wt%, respectively, and then stirred uniformly while maintaining the silver at 75 ° C. After a period of time, the micronized cellulose was gelled with increasing density to obtain a second salose gel. The obtained second cellulose gel was found to have a reduced fiber content, but the moisture content and density were similar to those of the first cellulose gel.

 6A and 6B are microstructure photographs of the first salose gel, and the surface of the first cell prepared by incubating at 30 ° C. for 14 days was lyophilized to observe the surface of the first gel. It can be seen that the first cellulose gel before pulverization constitutes a three-dimensional structure, as shown in the figure. The upper surface of the cellulose gel is in the form of a porous sheet in which nanofibers have a mesh shape, and the cross-section is Porosity

^'' NanoShi ¾ is layered and forms a three-dimensional structure connected to each other by fibers.

 I can see that there is.

 The microstructure of the second cell cellulose gel, which is pulverized at different times, is shown in FIGS.

 Shown in 7c. The grinding time was 5 minutes for the sample of FIG. 7A, 10 minutes for the sample pole of 7b, and 15 minutes for the sample of 7c. Using an electron microscope (S M, JSM-6400, JEOL, Japan) was confirmed at 10kV 10 ~ 12 mm distance conditions, each sample was measured by gold coating for 70 seconds using a sputter coater (Sputter coater). After freeze-drying the cellulose gel after grinding, the microstructure was confirmed, and it was confirmed that the nanofibers were broken. As the grinding time increased, the length of the nanofibers was shortened. It showed a net shape similar to, and the water content was similar as shown in Table 1 below. ，

[68] Table 1 [Table 1]

Moisture Content Comparison

[70] Apocrypha Example 2-Microbial Samloose Gem Mask Pack

 In order to prepare a microbial cell with mask gel, a citrus fruit was fermented and the primary microorganism salulose gel was pulverized with a mixer, grinder and homomixer to obtain micronized cellulose. The crushed gamgle fermented undifferentiated cell was weighed to 30 ^% in the total crude solution, placed in the main container, and the distilled water was quantitatively weighed (70 wt%) and mixed with the citrus fermented cell cellulose in the main container. Dispersion was dispersed for 20 minutes or more at 3000 rpm using a homomixer. At the time of dispersion, the main vessel was heated with a hot plate so that the temperature of the crude liquid was 60 ° C or higher.

 After the dispersion is completed, the crosslinking agent is quantitatively mixed with distilled water, and then introduced into a main container, followed by carrageenan, LBG (Locust Bean Gum), and xanthan gum.

(Xanthangum) and the like was mixed with glycerin (Glycerin) at room temperature. When the temperature of the undifferentiated salose mixed with distilled water was 60 ^o C or more, the natural polymer mixed with glycerin was added to the main container, and then mixed and dissolved in an azimixer for 10 minutes at 800 rpm. After dissolution was completed, preservatives, emulsifiers, fragrances, solubilizers, and the like were added, followed by further dissolution in the azimixer for 10 minutes at 800 rpm. Finally, defoaming was performed to complete the secondary cell cellulose gel.

 [73] The finished gel is coated on a transparent release paper for the purpose of exfoliation effect and product protection.

 Was carried out. The coating conditions were maintained at a temperature in the range of -5 ° C to the phase, the humidity was maintained at about 50%, and the coating thickness was maintained at about 0.5 to 1.2 mm.

 [0075] The coated second salose gel can be formed in various forms and can be pressed before or after gel coating. The second salose gel was pressed and molded into a facial shape to prepare a cosmetic mask pack (see FIG. 8). For comparison, the first salulose gelol before micronization was pressed and molded to prepare a mask pack of the same type (see FIG. 9). The obtained mask packs were almost identical in color, shape, physical strength and the like.

 [75] Also, a mask pack formed of a first cell with a rose gel and a second gelled powder after grinding

 Tensile strength of each of the mask packs formed of cellulose quality was tested. As shown in Table 2, the second cellulose gel was used as the mask pack.

Tensile strength was found to be sufficient for use, and it was confirmed that the tensile strength could be further improved depending on the level of compression. TABLE 2

 [Table 2]

Tensile Strength Comparison

 Thus, microbial salose gels prepared according to the present invention are characterized by low raw materials.

 It can be seen that it is very effective to manufacture a large amount of high quality mask pack even after being put.

 FIG. 10 illustrates a second cell prepared in the same method as another embodiment.

 By adding a functional material expressing the color was prepared a mask pack close to golden color. As such, the microorganism salulose gel according to the present invention can be utilized as a variety of cosmetic mask pack, it is effective to apply as a medical treatment by adding other additives.

 [8] The existing cosmetic mask pack mainly contains plant salose.

 It was prepared by impregnating a lotion with a nonwoven fabric or by applying collagen to a nonwoven fabric. Non-woven fabric in mask pack material is the most commonly used material or water-retaining property because of its simple molding and thickness control. On the other hand, the pharmaceutical image therapy can be prepared by impregnating the medical treatment gauze in a fibrous material of the medical treatment, in this case, in order to combine the image therapy to plentifully layered gauze.

 On the other hand, the microbial salulose gel according to the present invention is excellent in water retainability, excellent moldability, and physical strength, so it is necessary to use a separate nonwoven fabric to maintain its shape as in the conventional mask pack. There is no need to form a thick layered structure because it is easy to impregnate other materials even when used for medical purposes.

 [82] Bacterial cellulose derived from microorganisms has higher crystallinity and superior physical properties such as mechanical strength, adsorption, water retention, suspension stability, and binding properties compared to cellulose produced by ordinary plants, so that food additives, industrial materials, The microbial cell cellulose gel of the present invention is remarkably improved in the production yield of cellulose gel by pulverizing the first salose gel obtained through static culture followed by second gelation. Increase and accordingly microorganisms

 It will greatly improve the industrial availability of salolos gels.

 [83]

 The present invention has been exemplarily described through the preferred embodiments, but the present invention is not limited only to the specific embodiments as described above, but the scope of the technical idea and the scope of the claims of the present invention are specifically described. It may be modified, changed or improved in various forms within.

[85] Industrial availability

 The present invention is directed to a method for producing microorganism cells, which will be used in various chemical industries in connection with the cell-related industries.

Claims

Claim

Primary gelling agent by adding microbial strain to the culture medium and incubating 1  Preparing cellulose gel, The first sal was ground in a mixed state with water in a rose gel to obtain a finely divided sal, which was out of gel state, and the water was mixed with the finely divided cell, which had gone out of gel state, and an excipient was added. To a second gel by stirring at a temperature of 60 ^° C or more to prepare a second saloseogel.  The excipient is a microbial cell characterized in that it is a gelling agent selected from galactomannan, glucomannan, guar gum, locust bean gum, pluronic, agar, algin, carrageenan, xanthan gum, tara gum, tamarind gum, and gellan. Rose gel production method.  According to claim 1,  The method for producing a microbial sal loose gel further comprising the step of compressing the second cell with a rose gel.  In paragraph 2,  The method of claim 1, further comprising the step of slicing the second cell with a rose gel.

In paragraph 1,  The second cell cellulose gel is characterized in that the density of the fiber is lower than that of the first cellulosic gel.