High-yield bacterial cellulose strain and application thereof
By culturing Komagataeibacter nataicola JF11 strain in pre-fermented coconut water liquid medium, the problems of low yield and environmental sensitivity of Acetobacter xylinum strains were solved, achieving efficient and low-cost bacterial cellulose production, which is suitable for high-strength materials and other fields.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- HAINAN UNIV
- Filing Date
- 2026-01-14
- Publication Date
- 2026-05-12
AI Technical Summary
Existing Acetobacter xylinum strains have problems in bacterial cellulose production, including limited yield, sensitivity to fermentation environment, and high production cost, resulting in insufficient bacterial cellulose structure and performance.
The Komagataeibacter nataicola JF11 strain was cultured in pre-fermented coconut water liquid medium. By precisely matching the physiological characteristics of the strain, its metabolic pathways were activated, thereby improving the yield and production efficiency of bacterial cellulose.
It achieves efficient and low-cost bacterial cellulose production, with a simple cultivation process, high yield, and excellent bacterial cellulose structure and properties, making it suitable for high-strength materials and other fields.
Smart Images
Figure CN122012312A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of microbial technology, and more specifically, to a high-yield bacterial cellulosic strain and its applications. Background Technology
[0002] Bacterial cellulose (BC) is a natural polymer synthesized by microorganisms. It has unique properties such as high purity, high crystallinity, high water retention, good biocompatibility and biodegradability, and has broad application prospects in food (such as coconut jelly), medicine (such as artificial skin and drug carriers), cosmetics and advanced materials.
[0003] Currently, the main bacterial strain used for industrial production of bacterial cellulose is Acetobacter xylinum. However, Acetobacter xylinum has some inherent defects, such as: (1) limited bacterial cellulose production, resulting in high production costs; (2) being sensitive to fermentation environment (such as pH, dissolved oxygen, and inhibitors), and the fermentation process is unstable; (3) the structure and properties of the produced bacterial cellulose are still insufficient in certain specific applications (such as high-strength materials).
[0004] Based on this, a novel bacterial strain capable of stable and efficient BC synthesis in low-cost pre-fermented coconut water (FCW) liquid medium is provided, which is crucial for promoting the low-cost and high-performance application of bacterial cellulose. Summary of the Invention
[0005] In view of this, the present invention proposes a high-yield bacterial cellulosic strain and its application, aiming to solve at least one of the problems in the current background art.
[0006] This invention proposes a high-yield bacterial cellulosic strain, wherein the high-yield bacterial cellulosic strain is... Komagataeibacter nataicola JF11 is deposited at the Guangdong Provincial Center for Microbial Culture Collection, with accession number GDMCC NO:67317, and the deposit date is November 18, 2025.
[0007] The present invention also provides a microbial agent comprising the high-yield bacterial cellulosic strain described in the above technical solution.
[0008] The present invention also provides a composition comprising the high-yield bacterial cellulosic strain described in the above-described technical solution.
[0009] The present invention also provides an application of the high-yield bacterial cellulose strain as described in the above technical solution, wherein the application is in the production of bacterial cellulose.
[0010] The present invention also provides an application of the microbial agent as described in the above technical solution, wherein the application is in the production of bacterial cellulose.
[0011] The present invention also provides an application of the composition as described in the above technical solution, characterized in that the application is in the production of bacterial cellulose.
[0012] The present invention also provides a method for producing bacterial cellulose, comprising the following steps: (1) Preparation of culture medium: Prepare pre-fermented coconut water liquid culture medium. The formula of the pre-fermented coconut water (FCW) liquid culture medium is (NH4)2SO4 3g / L, MgSO4・7H2O 0.3g / L, KH2PO4 0.3g / L, pre-fermented coconut water 70%, sugar content adjusted to 6°Brix, pH 4.5, and sterilized at 115℃ for 25min; (2) Inoculation and culture: The high-yield bacterial cellulose strain is inoculated into the culture medium and cultured. (3) Harvesting and purification: After the static culture is completed, the bacterial cellulose membrane is collected, washed, and then dried at 70°C to constant weight to obtain the bacterial cellulose; The high-yield bacterial cellulose strain is the high-yield bacterial cellulose strain according to claim 1.
[0013] Preferably, the high-yield bacterial cellulosic strain described in step (2) is inoculated into the culture medium at a volume ratio of 2%.
[0014] Preferably, the washing process specifically involves: washing with deionized water, treating with 0.1 mol / L NaOH solution in a 75°C water bath for 2 hours, soaking in 0.5% acetic acid solution for 1 day, and rinsing with clean water.
[0015] Preferably, the culture in step (2) is specifically: static culture at 30°C for 4-8 days.
[0016] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention provides a high-yield bacterial cellulosic strain, namely... Komagataeibacter nataicola JF11, deposited at the Guangdong Provincial Center for Microbial Culture Collection (GDMCC NO:67317) on November 18, 2025, represents a high-yield bacterial cellulose strain. This invention provides a strain capable of efficiently producing bacterial cellulose with high yield, a simple cultivation process, and low production costs. Attached Figure Description
[0017] Various other advantages and benefits will become apparent to those skilled in the art upon reading the following detailed description of preferred embodiments. The accompanying drawings are for illustrative purposes only and are not intended to limit the invention. Furthermore, the same reference numerals denote the same parts throughout the drawings. In the drawings: Picture 1 This is a schematic diagram showing the yield results of Example 2 and Comparative Examples 1-4; Picture 2 The figures show the results of bacterial biofilm formation experiments in Example 2 and Comparative Examples 1-4; Picture 3 The graph shows the time-series changes in the activity of key enzymes in Example 2 and Comparative Examples 1-4. Detailed Implementation
[0018] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention. It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the present invention.
[0019] Furthermore, regarding the numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included within this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.
[0020] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.
[0021] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.
[0022] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.
[0023] This invention provides a high-yield bacterial cellulosic strain, wherein the high-yield bacterial cellulosic strain is... Komagataeibacter nataicola JF11 is deposited at the Guangdong Provincial Center for Microbial Culture Collection, with accession number GDMCC NO:67317, and the deposit date is November 18, 2025.
[0024] Preferably, the present invention also provides a method for culturing high-yield bacterial cellulosic strains, specifically: (1) Setting up the culture medium: Pre-fermented coconut water (FCW) liquid culture medium: (NH4)2SO4 3g / L, MgSO4•7H2O 0.3g / L, KH2PO4 0.3g / L, pre-fermented coconut water 70%, sugar content adjusted to 6°Brix, pH 4.5, sterilized at 115℃ for 25min; Fresh coconut water (NCW) liquid culture medium: (NH4)2SO4 3g / L, MgSO4•7H2O 0.3g / L, KH2PO4 0.3g / L, fresh coconut water 70%, sugar concentration adjusted to 6°Brix, pH 4.5, sterilized at 115℃ for 25min.
[0025] (2) Inoculation amount: Inoculate the high-yield bacterial cellulose strain into cooled pre-fermented coconut water (FCW) liquid medium or fresh coconut water (NCW) liquid medium at a ratio of 2% (v / v).
[0026] (3) Culture conditions: static culture at 30℃ for 6 days.
[0027] (4) Harvesting and determination: After the culture is completed, collect the bacterial cellulose membrane, wash it with deionized water, soak it in 0.1 mol / L NaOH solution, and water bath at 75℃ for about 2 hours. When the bacterial cellulose membrane turns milky white, take it out and soak it in 0.5% acetic acid solution for 1 day. Finally, rinse the bacterial cellulose membrane repeatedly with clean water. Dry it at 70℃ to constant weight and weigh the dry weight.
[0028] The natural components (such as sugars and amino acids) in the pre-fermented coconut water (FCW) liquid culture medium selected in this invention can specifically activate the JF11 metabolic pathway. Specifically, through the pre-fermentation process, the main carbon sources (sucrose and glucose) in fresh coconut water that easily trigger metabolic shunting are efficiently converted into lactic acid, ethanol, and acetic acid, which are preferred by *Codonopsis cocovenenans*. Therefore, the pre-fermented coconut water (FCW) liquid culture medium is precisely matched to the physiological characteristics of *Codonopsis cocovenenans*, maximizing its metabolic flow towards bacterial cellulose synthesis, thereby achieving a specific increase in bacterial cellulose production. This invention does not specifically limit the source of each component in the culture medium; any source well known to those skilled in the art can be used.
[0029] The present invention also provides a microbial agent comprising the high-yield bacterial cellulosic strain described in the above technical solution.
[0030] The present invention also provides a composition comprising the high-yield bacterial cellulosic strain described in the above-described technical solution.
[0031] The present invention also provides an application of the high-yield bacterial cellulose strain as described in the above technical solution, wherein the application is in the production of bacterial cellulose.
[0032] The present invention also provides an application of the microbial agent as described in the above technical solution, wherein the application is in the production of bacterial cellulose.
[0033] The present invention also provides an application of the composition as described in the above technical solution, characterized in that the application is in the production of bacterial cellulose.
[0034] Example 1 Screening of strains (1) Seed liquid for production was purchased from a coconut fiber (bacterial cellulose) production plant in Haikou City, Hainan Province. The seed liquid was diluted tenfold and spread on a pre-fermented coconut water (FCW) solid medium for separation. Single colonies were picked for liquid fermentation verification. Those that could produce film were BC-producing bacteria. Multiple pure strains were obtained through multiple purifications. (2) Screening was conducted using BC yield as the criterion to identify high-yielding bacterial cellulosic strains. Komagataeibacter nataicola JF11; (3) Strain identification: The strain was identified as *Colocasia esculenta* by 16S rRNA gene sequence analysis.
[0035] Example 2 High-yield bacterial cellulose strain Komagataeibacter nataicola JF11 cultivation (1) Preparation of pre-fermented coconut water Select fresh coconuts of uniform maturity, clean the shells, open them with a sterile knife, collect the coconut water, filter it through double-layer sterile gauze to remove suspended fruit pulp particles, and obtain clear coconut water with an initial sugar content of about 4.0 and an initial pH of 5-6. Place it in the laboratory for natural fermentation for 1-7 days until the pH stabilizes at 3.5-4.0 and the sugar content stabilizes at about 2.0, then stop the fermentation and store it at -20℃ for later use.
[0036] (2) Preparation of pre-fermented coconut water (FCW) liquid culture medium Add 200 mL of deionized water to a sterile beaker, then add 3.000 g of (NH4)2SO4, 0.300 g of MgSO4・7H2O, and 0.300 g of KH2PO4 in sequence. Stir with a magnetic stirrer until completely dissolved to form a salt mother liquor. Pour 700 mL of refrigerated pre-fermented coconut water into a 1 L volumetric flask, add the above-mentioned salt stock solution, and stir well; adjust the sugar content of the mixture with sucrose powder, adding 1 g of sucrose and stirring until dissolved, then measuring with a saccharimeter until the sugar content accurately reaches 6.0°Brix; slowly add 0.1 mol / L hydrochloric acid dropwise to the culture medium, while monitoring the pH in real time with a pH meter, and accurately adjust the pH to 4.5 ± 0.05; Dilute the culture medium to 1L with deionized water, stir well, and dispense 50mL into 100mL Erlenmeyer flasks. Place the dispensed culture medium into an autoclave, set the temperature to 115℃ and the pressure to 0.07MPa, and autoclave for 25min. After sterilization, allow the pressure to release naturally to atmospheric pressure, remove the medium, and place it on a sterile workbench to cool to room temperature for later use.
[0037] (3) Preparation of fresh coconut water (NCW) liquid culture medium Except for using 700 mL of fresh coconut water instead of pre-fermented coconut water, the other salt ratios, sugar content adjustments, pH adjustments, volumetric dispensing, and sterilization conditions were the same as those for pre-fermented coconut water (FCW) liquid culture medium. (4) Activation of strain A small amount of JF11 bacterial culture was picked from the preservation tube and inoculated onto pre-fermented coconut water (FCW) solid medium using the streak plating method. Place the inoculated pre-fermented coconut water (FCW) solid medium in a 30°C incubator and invert it for 3-7 days until uniform, smooth, milky-white colonies form on the surface of the medium. Use a sterile inoculation loop to pick a single colony from the medium and transfer it to 5 ml of pre-fermented coconut water (FCW) liquid medium for activation for 48 hours. Then, inoculate 2% of this liquid medium into an Erlenmeyer flask containing 50 ml of pre-fermented coconut water (FCW) liquid medium. Place the flask in an incubator at 30°C and incubate statically for 48 hours, observing the bacterial culture every 8 hours. When the OD of the bacterial culture reaches a certain level... 600 When the value reaches around 0.6 (a film appears), keep it for later use.
[0038] Using a sterile pipette, aspirate the seed culture and inoculate it at a ratio of 2% (v / v) into pre-fermented coconut water (FCW) liquid medium cooled to room temperature or fresh coconut water (NCW) liquid medium. After inoculation, gently invert the Erlenmeyer flask 3-5 times to ensure thorough mixing of the seed culture with the medium. The inoculated Erlenmeyer flasks were placed in a constant temperature incubator at 30℃±0.5℃ for 6 days. After incubation, the bacterial cellulose membrane was collected and washed with deionized water. Then, the cellulose membrane was soaked in 0.1mol / L NaOH solution and incubated at 75℃ for about 2 hours. Once the membrane turned milky white, it was removed and soaked in 0.5% acetic acid solution for 1 day. Finally, the membrane was repeatedly rinsed with clean water. It was then dried at 70℃ to constant weight, and the dry weight of bacterial cellulose BC was measured.
[0039] Comparative Example 1 All other steps are the same as in Example 2, the only difference being that the cultured bacterial strain is... komagataeibacter nataicola JG15.
[0040] Comparative Example 2 All other steps are the same as in Example 2, the only difference being that the cultured bacterial strain is... Komagataeibacter nataicola JY831.
[0041] Comparative Example 3 All other steps are the same as in Example 2, the only difference being that the cultured bacterial strain is... Komagataeibacter nataicola JY6211.
[0042] Comparative Example 4 The other steps are the same as in Example 2, the only difference being that the cultured bacterial strain is ( Novacetimonas cocois) WE7.
[0043] The strains of Comparative Examples 1 to 3 were all isolated and purified from the production seed liquid of a coconut fiber (bacterial cellulose) production plant in Haikou City, Hainan Province, while the strain of Comparative Example 4 was isolated and purified from deteriorated coconut milk fermentation product.
[0044] Performance testing (1) The yields of bacterial cellulose (BC) obtained in Example 2 and Comparative Examples 1-4 were statistically analyzed, and the results are as follows: Picture 1 As shown, based on Picture 1 It can be seen that the bacterial cellulose yield obtained in Example 2 was the highest.
[0045] (2) JF11 was inoculated into pre-fermented coconut water (FCW) liquid medium and fresh coconut water (NCW) liquid medium. The BC yield results are as follows: Picture 2 As shown, based on Picture 2 It can be seen that the BC yield in pre-fermented coconut water (FCW) liquid medium is significantly higher than that in fresh coconut water (NCW) liquid medium.
[0046] (3) Based on Picture 2 It was found that the inflection points for BC yield growth were observed on days 2, 4, and 6 of JF11 inoculation in pre-fermented coconut water (FCW) liquid medium and fresh coconut water (NCW) liquid medium. These three inflection points were used to measure uridine diphosphate glucose pyrophosphorylase (UDP-). glucose pyrophosphosphprylase The content of UGP was as follows: Picture 3 As shown, based on Picture 3 It can be seen that JF11 exhibits high UGP enzyme activity in pre-fermented coconut water (FCW) liquid medium. This enzyme is a key rate-limiting enzyme in the bacterial cellulose synthesis pathway, and its increased activity directly promotes BC synthesis.
[0047] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit it. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the specific implementation of the present invention. Any modifications or equivalent substitutions that do not depart from the spirit and scope of the present invention should be covered within the scope of protection of the claims of the present invention.
Claims
1. A high-yield bacterial cellulose strain, characterized in that, The high-yield bacterial cellulose strain is Komagataeibacter nataicola JF11 is deposited at the Guangdong Provincial Center for Microbial Culture Collection, with accession number GDMCC NO:67317, and the deposit date is November 18, 2025.
2. A microbial agent, characterized in that, The bacterial agent includes the high-yield bacterial cellulose strain as described in claim 1.
3. A composition, characterized in that, The composition comprises the high-yield bacterial cellulose strain of claim 1.
4. The application of a high-yield bacterial cellulosic strain as described in claim 1, characterized in that, The application is in the production of bacterial cellulose.
5. The application of the microbial agent as described in claim 2, characterized in that, The application is in the production of bacterial cellulose.
6. An application of the composition as described in claim 3, characterized in that, The application is in the production of bacterial cellulose.
7. A method for producing bacterial cellulose, characterized in that, Includes the following steps: (1) Preparation of culture medium: Prepare pre-fermented coconut water (FCW) liquid culture medium. The formula of the pre-fermented coconut water (FCW) liquid culture medium is (NH4)2SO4 3g / L, MgSO4・7H2O 0.3g / L, KH2PO4 0.3g / L, pre-fermented coconut water 70%, sugar content adjusted to 6°Brix, pH 4.5, and sterilized at 115℃ for 25min; (2) Inoculation and culture: The high-yield bacterial cellulose strain is inoculated into the culture medium and cultured. (3) Harvesting and purification: After the static culture is completed, the bacterial cellulose membrane is collected, washed, and then dried at 70°C to constant weight to obtain the bacterial cellulose; The high-yield bacterial cellulose strain is the high-yield bacterial cellulose strain according to claim 1.
8. The method for producing bacterial cellulose according to claim 7, characterized in that, The high-yield bacterial cellulose strain described in step (2) is inoculated into the culture medium at a volume ratio of 2%.
9. The method for producing bacterial cellulose according to claim 7, characterized in that, The washing process specifically involves: washing with deionized water, treating with 0.1 mol / L NaOH solution in a 75°C water bath for 2 hours, soaking in 0.5% acetic acid solution for 1 day, and rinsing with clean water.
10. The method for producing bacterial cellulose according to claim 7, characterized in that, The specific culture described in step (2) is: static culture at 30℃ for 4-8 days.