Vibrio parahaemolyticus cj5 and applications thereof

Abstract

The application discloses a Vibrio parahaemolyticus CJ5 and application thereof, and belongs to the technical field of microorganisms. The Vibrio parahaemolyticus CJ5 provided by the application is separated and screened from workshop water of a sea cucumber breeding base, the strain can efficiently and directionally degrade SC-FUC, the degradation product has a relatively high sulfate retention rate and a relatively concentrated molecular mass distribution, under the conditions that the growth temperature is 30 DEG C, the salinity is 3%, the pH value is 8.0 and the inoculation amount is 3%, the degradation rate of the strain to SC-FUC reaches 61.47%, the sulfate mass fraction of the degradation product is 34.55%, and the relative molecular mass distribution range is within 800 Da, so that the sea cucumber fucoidan with DP 2-5 can be prepared.

Description

Technical Field

[0001] This invention relates to a bacterium and its application, specifically to Vibrio parahaemolyticus CJ5 and its application in the degradation of sea cucumber fucose sulfate, belonging to the field of microbial technology. Background Technology

[0002] Fucoidan (FUC) is a polysaccharide compound rich in sulfated fucoidan, isolated from brown algae and echinoderms. Studies have shown that sea cucumber-derived fucoidan (also known as sea cucumber fucoidan sulfate, abbreviated as SC-FUC) has a regular linear structure. Compared to seaweed-derived fucoidan (SW-FUC), SC-FUC has a simpler structure, giving it unique advantages in both structure and function. SC-FUC exhibits various biological activities, including antiviral, antioxidant, anticoagulant, antitumor, and anti-inflammatory effects. However, the large molecular weight, high viscosity, and poor absorption of naturally occurring SC-FUC severely limit its application in pharmaceuticals, food, and cosmetics.

[0003] Currently, the main methods for degrading fucoidan are chemical methods, including acid hydrolysis and oxidation. However, chemical methods suffer from drawbacks such as stringent production conditions, inconsistent degrees of polymerization in the products, difficulty in purification, and low yields, thus limiting their widespread industrial application.

[0004] Biodegradation methods offer mild reaction conditions and high specificity, but research on the preparation of low-molecular-weight sea cucumber fucoidan oligosaccharides (SC-FO) through microbial degradation of SC-FUC is still in its early stages. Existing technologies face the following prominent problems and obvious limitations:

[0005] (1) Lack of dedicated microbial resources and superior strains capable of efficiently degrading SC-FUC;

[0006] (2) During the microbial degradation process, a large number of key active groups such as sulfate in SC-FUC are lost, the retention rate of active groups in the degradation products is low, and the relative molecular mass distribution of the degradation products is not concentrated.

[0007] Therefore, it is urgent to screen strains that can efficiently and directionally degrade SC-FUC and ensure that the degradation products have a high retention rate of active groups and a very concentrated molecular weight distribution. This will fill the gap in marine microbial resources for degrading SC-FUC, lay the foundation for developing industrial production processes of highly active fucoidan, and have important application value for the development of marine pharmaceuticals, functional foods, and cosmetics industries.

[0008] Chinese invention patent CN 106635920A discloses a bacterium capable of producing high levels of fucoidan-degrading enzymes—Pseudomonas alterniflora 0U03 strain. However, the fucoidan-degrading enzymes produced by this strain are directed to degrade algal-derived fucoidan (SW-FUC). Since SW-FUC and SC-FUC have different structures, it is unknown whether Pseudomonas alterniflora 0U03 strain can simultaneously and directedly degrade SC-FUC. Furthermore, the degradation rate of fucoidan and the sulfate retention rate of the degradation products have not been reported.

[0009] The existing paper, "Screening, Identification, Fermentation Condition Optimization and Enzymatic Properties Study of Fucoidan Sulfate-Producing Bacteria from Sea Cucumber," discloses a bacterium capable of degrading fucoidan (SC-FUC) from sea cucumber. Flavobacteriaceae .sp HDJ3, however, SC-FUC after Flavobacteriaceae The relative molecular mass distribution and sulfate retention rate of the degradation products after HDJ3 degradation were not reported. Summary of the Invention

[0010] To address the shortcomings of existing technologies, the present invention aims to provide a strain capable of efficiently and directionally degrading sea cucumber-derived fucoidan (SC-FUC) while ensuring that the degradation products have a high retention rate of active groups and a relatively concentrated molecular weight distribution.

[0011] To achieve the above objectives, the present invention adopts the following technical solution:

[0012] Vibrio parahaemolyticus CJ5, Latin name is Vibrio parahaemolyticus CJ5 is deposited at the China General Microbiological Culture Collection Center (CGMCC) on June 11, 2025, with accession number CGMCCNo.34858, located in Beijing, China.

[0013] The aforementioned application of Vibrio parahaemolyticus CJ5 in the degradation of sea cucumber fucose sulfate.

[0014] The advantages of this invention are as follows: The Vibrio parahaemolyticus CJ5 strain provided by this invention is isolated and screened from the workshop water of a sea cucumber farming base. This strain can efficiently and directionally degrade SC-FUC, and the degradation products have a high sulfate retention rate and a relatively concentrated molecular weight distribution. Under the conditions of growth temperature of 30℃, salinity of 3%, pH of 8.0, and inoculum size of 3%, the degradation rate of SC-FUC by this strain reached 61.47%, the sulfate mass fraction of the degradation products was 34.55%, and the relative molecular weight distribution range was within 800 Da, which can be used to prepare sea cucumber fucoidan with DP 2-5. Attached Figure Description

[0015] Figure 1 This is a colony morphology diagram of strain CJ5;

[0016] Figure 2 This is a diagram of the cell shape of strain CJ5;

[0017] Figure 3 This is a phylogenetic tree diagram of strain CJ5;

[0018] Figure 4 This is a graph showing the hemolysis test results for strain CJ5;

[0019] Figure 5 This is a graph showing the growth results of strain CJ5 at different culture temperatures;

[0020] Figure 6 This is a graph showing the growth results of strain CJ5 at different sodium chloride concentrations;

[0021] Figure 7 This is a graph showing the growth results of strain CJ5 at different pH values;

[0022] Figure 8 This is a chromatogram of PMP derivatives of monosaccharide standards;

[0023] Figure 9 This is a chromatogram of the PMP derivative of SC-FO;

[0024] Figure 10 This is the HPSEC map of SC-FO. Detailed Implementation

[0025] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments.

[0026] I. Sample Collection

[0027] In July 2023, multiple water samples were collected from the sea cucumber farming base of Yantai Anyuan Aquatic Products. The collected water was filtered through a sterile filter membrane, and the filter membrane was placed into 100mL sterile centrifuge tubes. The tubes were then sealed with sealing film, placed in ice boxes, and transported to the laboratory for later use.

[0028] II. Preparation of Sea Cucumber-Derived Fucoidan (SC-FUC)

[0029] The dried sea cucumber was pulverized and added to acetone for defatting at 4°C. After 24 hours, the acetone was evaporated, and a 0.1 mol / L sodium acetate buffer solution (pH 6.0, containing 10 g / L papain, 5 mmol / L EDTA and 5 mmol / L cysteine) was added at a material-to-liquid ratio of 1 g:30 mL. The mixture was continuously enzymatically hydrolyzed in a 60°C water bath for 24 hours. After centrifugation (4000 rpm, 20 min, 20°C), the supernatant was collected, and hexadecylpyridine chloride (final concentration 5 g / L) was added. After standing for 24 hours, the mixture was centrifuged again (4500 rpm, 15 min, 20°C), and the precipitate was collected.

[0030] A 3 mol / L sodium chloride solution and ethanol were mixed at a volume ratio of 20:3 to obtain a mixed solvent. The collected precipitate was dissolved in the mixed solvent and then purified by precipitation with 95% (v / v) ethanol. After that, it was washed with a gradient of ethanol (80% (v / v) and 95% (v / v)) and finally dried in a constant temperature drying oven at 60℃ to obtain SC-FUC crude sugar.

[0031] To eliminate interference from proteins and other structural polysaccharides in inducing enrichment and screening of bacterial strains, the obtained crude SC-FUC was further purified. Specifically, the crude SC-FUC was separated and purified using a DEAE Sepharose Fast Flow anion exchange column. Gradient elution was performed sequentially using distilled water, 0.2 mol / L sodium chloride solution, and 0.6 mol / L sodium chloride solution. The 0.6 mol / L sodium chloride elution fraction was analyzed to have the following characteristics: protein content less than 1% (w / w), relative molecular mass approximately 1570 kDa, fucose content exceeding 90% in the monosaccharide composition, and sulfate content exceeding 35%. This fraction was desalted and concentrated using an ultrafiltration membrane with a molecular weight cutoff of 1000 Da. The ultrafiltration retentate was collected and freeze-dried under vacuum to obtain a powdered high-purity SC-FUC sample.

[0032] III. Microbial strain enrichment, screening, isolation and purification

[0033] 1. Culture medium preparation

[0034] Liquid culture medium (w / v): SC-FUC 0.2% (sole carbon source), dipotassium hydrogen phosphate 0.1%, ammonium sulfate 0.2%, magnesium sulfate 0.1%, calcium chloride 0.01%, ferrous sulfate 0.01%, sodium chloride 3%, pH adjusted to 7.0.

[0035] Solid culture medium: Add 1.5% (w / v) agar powder to the liquid culture medium.

[0036] 2. Induced enrichment of strains

[0037] Add 5 mL of ddH2O to each sample tube. Take 2.0 mL of each collected sample and inoculate it into 50 mL of liquid culture medium with SC-FUC as the sole carbon source. Incubate at 25 °C and 180 rpm with shaking. After each enrichment culture for 5 days, transfer a portion of the culture medium to fresh liquid culture medium. Repeat this process for three consecutive rounds to gradually increase the relative abundance of the target strain, which is beneficial for the subsequent isolation and identification of the target strain.

[0038] 3. Initial screening of bacterial strains

[0039] The bacterial culture, after three rounds of enrichment, was serially diluted (10⁻⁶). -3 -10 -5 Then, 0.5 mL of the diluted solution was spread onto the surface of a solid culture medium using SC-FUC as the sole carbon source and incubated upside down at 25°C for 5 days. After incubation, all single colonies with significant morphological differences on the culture medium were observed and recorded. Typical single colonies were selected for three consecutive streak plate purifications. The purified strains were inoculated onto slant agar and incubated at 25°C until the bacterial growth was sufficient. Then, they were temporarily stored at 4°C for later use.

[0040] After the culture was completed, a total of 7 single colonies with obvious morphological differences were obtained, which were numbered and named with CJ (CJ1, CJ2, CJ3, CJ4, CJ5, CJ6, CJ7) and entered the next round of screening as candidate strains.

[0041] 4. Secondary screening of bacterial strains

[0042] The seven strains obtained from the initial screening were inoculated into 50 mL test tubes containing 15 mL of liquid culture medium, and then cultured at 25 °C and 180 rpm with shaking for 3 days. At 24 h, 36 h, 48 h, 60 h, and 72 h of culture, the remaining SC-FUC content in the culture medium was determined using the phenol-sulfuric acid method, and the degradation rate of SC-FUC by each strain was calculated accordingly. Finally, strains with a degradation rate exceeding 20% ​​were identified as candidate target bacteria for further research.

[0043] The degradation rates of SC-FUC by each strain at different culture times are shown in Table 1.

[0044] Table 1. Degradation rate of SC-FUC by various strains at different culture times

[0045]

[0046] Experimental results showed that after further cultivation, the seven strains exhibited different SC-FUC utilization abilities. Among them, strain CJ5 was the first to exceed 20% degradation rate of SC-FUC after 24 hours of cultivation, while the degradation rates of other strains did not reach 20% after the end of cultivation. Therefore, strain CJ5 was preliminarily identified as a candidate strain for efficient targeted degradation of SC-FUC.

[0047] IV. Colony morphology, cell shape, and physiological and biochemical characteristics of strain CJ5

[0048] 1. Colony morphology

[0049] Under aseptic conditions, strain CJ5 was inoculated onto a solid culture medium using the three-line method. After being cultured at 25°C until the bacterial colony was fully grown, it was temporarily stored at 4°C for later use.

[0050] The growth status of strain CJ5 on solid medium is shown in the figure. Figure 1 It was observed that strain CJ5 formed white, smooth, round colonies on the culture medium.

[0051] 2. Cell shape

[0052] Strain CJ5 was inoculated onto solid culture medium and cultured at 25°C for 24 h. The bacterial cells were then scraped from the solid culture medium into centrifuge tubes and washed three times with Tris-HCl buffer (pH 7.2-7.4) for 10 min each time. The cells were then fixed with 2.5% (w / v) glutaraldehyde solution for 3 h. After fixation, the cells were washed three more times with Tris-HCl buffer (pH 7.2-7.4) for 10 min each time, followed by two rinses with sterile pure water. The strain was then subjected to a gradient dehydration process using 20% ​​(v / v), 40% (v / v), 60% (v / v), 70% (v / v), and 80% (v / v) ethanol solutions, with each concentration treated for 15 min. Finally, the strain was dehydrated twice with anhydrous ethanol to completely remove water. After dehydration, the sample was immersed in a 1:1 mixture of tert-butanol and ethanol for 15 min, and then transferred to pure tert-butanol for storage. Finally, the sample was dropped onto a clean coverslip, pre-cooled at -80°C, then vacuum dried, and finally observed on an instrument.

[0053] Electron microscopy results of strain CJ5 are shown in the figure. Figure 2It was observed that strain CJ5 was a smooth, short rod-shaped organism, approximately 0.3 μm-0.5 μm wide and 1.0 μm-1.5 μm long, and possessed a single flagellum.

[0054] 3. Physiological and biochemical characteristics

[0055] Physiological and biochemical identification revealed that strain CJ5 is a Gram-negative bacterium and exhibits the following characteristics:

[0056] (1) β-N-acetylglucosidase (BNAG) was positive (+), indicating that strain CJ5 may have the potential to degrade complex polysaccharides containing N-acetylglucosamine structures (such as chitin or sulfated polysaccharides);

[0057] (2) Both α-galactosidase (AGAL) and β-galactosidase (BGAL) were negative (-), indicating that strain CJ5 has a weak ability to break down galactosidic bonds.

[0058] (3) It can utilize glucose, mannose, rhamnose and lactose, indicating that strain CJ5 can efficiently metabolize these sugars, indirectly supporting the possibility that strain CJ5 may have the ability to obtain carbon sources by degrading polysaccharides.

[0059] V. Species Identification of Strain CJ5

[0060] The 16S rRNA gene is a conserved DNA sequence encoding bacterial ribosomal RNA (rRNA), widely distributed in the genomes of various bacteria. After sequencing, the predicted 16S rRNA sequences were compared with the NCBI 16S rRNA database using BLAST, with the parameter `identify>95`. Then, the top 30 16S rRNA sequences with the highest `identify` were selected (all sequences were included if insufficient), and multiple sequence alignment and splicing were performed using MAFFT software. Finally, a phylogenetic tree of strain CJ5 was constructed using FastTree software. The results are shown below. Figure 3 .

[0061] Comparison using the BLAST online program in the NCBI database revealed that strain CJ5 was similar to... Vibrio parahaemolyticus Because the sequence similarity of DH076 (Vibrio parahaemolyticus) reached 95%, strain CJ5 was classified as a Vibrio strain and named Vibrio parahaemolyticus CJ5, with the Latin name [missing information]. Vibrio parahaemolyticus CJ5.

[0062] VI. Detection of hemolysis by Vibrio parahaemolyticus CJ5

[0063] Take 1 μL of Vibrio parahaemolyticus CJ5 bacterial suspension and inoculate it onto a blood agar plate. After incubating at 37℃ for 24 h, observe the hemolysis results.

[0064] The hemolysis test results of Vibrio parahaemolyticus CJ5 are shown in the figure. Figure 4 .Depend on Figure 4 It can be seen that Vibrio parahaemolyticus CJ5 did not show a clear hemolytic zone on my wife's blood agar, and the Kanagawa test was negative. This indicates that Vibrio parahaemolyticus CJ5 is non-virulent.

[0065] VII. Growth of Vibrio parahaemolyticus CJ5 under different culture temperatures, sodium chloride concentrations, and pH conditions.

[0066] 1. Detection of Vibrio parahaemolyticus CJ5 growth at different culture temperatures

[0067] Vibrio parahaemolyticus CJ5 was inoculated into a 50 mL test tube containing 15 mL of liquid culture medium and cultured at 25 °C and 180 rpm for 12 h with shaking. Subsequently, 5% (v / v) inoculum was transferred to a 250 mL Erlenmeyer flask containing 50 mL of liquid culture medium. 200 μL of the sample was added to a 96-well plate, and the initial OD of the culture medium was measured using a microplate reader. 600nm Then, the samples were cultured at 20℃, 25℃, 30℃, 35℃, and 40℃ for 24 h with shaking. After the culture was completed, 200 μL of sample was added to a 96-well plate, and the OD of the culture medium was measured using a microplate reader after 24 h of culture. 600nm The growth rate of the strain was measured by the OD value of the culture medium after 24 hours of culture. 600nm OD of the culture medium at the beginning of the experiment 600nm The difference (△OD) 600nm () indicates. Each experimental group has 3 parallels.

[0068] The results of the growth detection of Vibrio parahaemolyticus CJ5 at different culture temperatures are shown in the figure. Figure 5 .Depend on Figure 5 It is known that Vibrio parahaemolyticus CJ5 can grow at temperatures ranging from 20℃ to 40℃, with the optimal growth temperature being 30℃.

[0069] 2. Detection of Vibrio parahaemolyticus CJ5 growth under different sodium chloride concentrations

[0070] 500 μL of Vibrio parahaemolyticus CJ5 culture cultured to the exponential phase was inoculated into 50 mL of liquid culture medium with sodium chloride concentrations (w / v) of 1%, 2%, 3%, 4%, and 5%, respectively. 200 μL of the sample was added to each well of a 96-well plate, and the OD value of the culture medium at the initial stage of the experiment was measured using a microplate reader. 600nm Then, the sample was shaken and cultured at 30°C for 24 hours at 150 rpm. After the culture was completed, 200 μL of sample was added to a 96-well plate, and the OD of the culture medium was measured using a microplate reader after 24 hours of culture. 600nm The growth rate of the strain was measured by the OD value of the culture medium after 24 hours of culture. 600nm OD of the culture medium at the beginning of the experiment 600nmThe difference (△OD) 600nm () indicates. Each experimental group has 3 parallels.

[0071] The results of the growth detection of Vibrio parahaemolyticus CJ5 under different sodium chloride concentrations are shown in the figure. Figure 6 .Depend on Figure 6 It is known that Vibrio parahaemolyticus CJ5 grows in a salinity range of 1%-5%, with an optimal salinity of 3%.

[0072] 3. Detection of Vibrio parahaemolyticus CJ5 growth at different pH values

[0073] 500 μL of Vibrio parahaemolyticus CJ5 culture cultured to the exponential phase was inoculated into 50 mL of liquid culture medium with pH values ​​of 5.0, 6.0, 7.0, 8.0, and 9.0, respectively. 200 μL of the sample was added to a 96-well plate, and the OD value of the culture medium at the initial stage of the experiment was measured using a microplate reader. 600nm Then, the sample was incubated on a shaker at 150 rpm at 30°C for 24 h. After incubation, 200 μL of sample was added to a 96-well plate, and the OD of the culture medium was measured using a microplate reader after 24 h of incubation. 600nm The growth rate of the strain was measured by the OD value of the culture medium after 24 hours of culture. 600nm OD of the culture medium at the beginning of the experiment 600nm The difference (△OD) 600nm () indicates. Each experimental group has 3 parallels.

[0074] The results of the growth of Vibrio parahaemolyticus CJ5 at different pH values ​​are shown in the figure. Figure 7 .Depend on Figure 7 It can be seen that the pH range for Vibrio parahaemolyticus CJ5 growth is 5.0-9.0, with the optimal growth pH being 8.0.

[0075] In summary, Vibrio parahaemolyticus CJ5 can grow in environments ranging from 20℃ to 40℃, with pH values ​​of 5.0 to 9.0 and sodium chloride concentrations (w / v) of 1% to 5%. The optimal growth temperature is 30℃, the optimal salinity is 3%, and the optimal pH is 8.0. Therefore, Vibrio parahaemolyticus CJ5 exhibits a wide optimal temperature range and strong tolerance to salt and acids / alkalis.

[0076] 8. Effect of different inoculum amounts on the degradation rate of SC-FUC

[0077] Based on the optimized culture conditions for Vibrio parahaemolyticus CJ5, the pH of the liquid culture medium was adjusted to 8.0, and the culture was carried out at 30℃. Vibrio parahaemolyticus CJ5 was inoculated into liquid culture medium containing SC-FUC at inoculum concentrations of 0.5%, 1%, 2%, 3%, 4%, 5%, and 6%, with three replicates per group. Samples were collected after 36 hours of incubation on a shaker at 150 rpm, and the degradation rate of SC-FUC was determined using the phenol-sulfuric acid method.

[0078] The degradation rate of SC-FUC at different inoculum amounts is shown in Table 2.

[0079] Table 2 Degradation rate of SC-FUC at different inoculum amounts

[0080]

[0081] As shown in Table 2, the degradation rate of SC-FUC gradually increases with the increase of inoculum amount. When the inoculum amount is 3%, the degradation rate reaches the highest level of 61.47%. Further increasing the inoculum amount will decrease the degradation rate of SC-FUC.

[0082] It is evident that the optimal inoculum size for Vibrio parahaemolyticus CJ5 to degrade SC-FUC is 3%, with a polysaccharide degradation rate exceeding 60%, demonstrating a good SC-FUC degradation effect.

[0083] IX. Analysis of Degradation Products

[0084] Liquid samples were collected after fermentation for 36 hours under optimal culture conditions. The samples were sterilized at 121℃ for 20 minutes, centrifuged (4000 rpm, 10 minutes, 4℃), and the supernatant was collected. Anhydrous ethanol was added at a volume ratio of 1:3, and the samples were refrigerated at 4℃ overnight. The samples were centrifuged again (4000 rpm, 10 minutes, 4℃), and the supernatant was collected. The supernatant was concentrated to 1 / 5 of the original volume by rotary evaporation. The supernatant was then desalted by dialysis using a dialysis bag with a molecular weight cutoff of 200 Da. After freeze-drying, sea cucumber fucoidan (SC-FO) was prepared and used to determine its monosaccharide composition, sulfate content, and relative molecular mass distribution.

[0085] 1. Monosaccharide composition

[0086] The monosaccharide composition of the monosaccharide standards and SC-FO was determined by pre-column derivatization-high performance liquid chromatography (PMP). The results are shown in [Figure number missing]. Figure 8 and Figure 9 .

[0087] Depend on Figure 8 and Figure 9 It can be seen that SC-FO is composed of three monosaccharides: fuc, glucuronic acid (GlcUA), and galactosamine (GalN), with a molar ratio of Fuc:GlcUA:GalN = 1.00:0.04:0.03. This indicates that SC-FO is mainly composed of fuc, with trace amounts of glucuronic acid and galactosamine.

[0088] 2. Sulfate content (mass fraction)

[0089] The sulfate content of SC-FO was determined using the barium chloride-gelatin turbidimetric method. The results showed that the sulfate mass fraction of SC-FO was 34.55%. This indicates that the degradation of SC-FO by Vibrio parahaemolyticus CJ5 effectively preserves important bioactive groups such as sulfate, resulting in SC-FO with high sulfate content.

[0090] 3. Relative molecular mass distribution

[0091] The relative molecular mass distribution of SC-FO was determined using high performance gel size exclusion chromatography (HPSEC). The specific chromatographic conditions are as follows:

[0092] (1) Chromatograph: Agilent 1260 high performance liquid chromatograph;

[0093] (2) Chromatographic column: TSK-gel G4000 PWXL (id 7.8mm×30.0cm);

[0094] (3) Detector: Differential Detector (RID);

[0095] (4) Mobile phase: 0.2 mol / L sodium chloride solution;

[0096] (5) Flow rate: 0.5 mL / min;

[0097] (6) Column temperature: 40°C;

[0098] (7) Standard products: Dextran series standard products;

[0099] (8) Molecular weight distribution calculation: Agilent GPC software was used for calculation.

[0100] The relative molecular mass determination results of SC-FO are shown in the figure. Figure 10 .Depend on Figure 10 It can be seen that the HPSEC peak of SC-FO is symmetrical and uniformly distributed. Through calculation, the relative molecular mass distribution range of SC-FO is within 800 Da. SC-FO with a degree of polymerization (DP) of 2-5 can be prepared, and the relative molecular mass distribution is very concentrated.

[0101] The above results indicate that Vibrio parahaemolyticus CJ5 has a highly efficient and excellent biodegradation effect on SC-FUC, and can be used to prepare DP2-5 sea cucumber fucoidan oligosaccharide.

[0102] IX. Preservation of Microbial Strains

[0103] Vibrio parahaemolyticus CJ5 (Latin name: Vibrio parahaemolyticusCJ5 has been deposited with the China General Microbiological Culture Collection Center (CGMCC) on June 11, 2025, with accession number CGMCCNo.34858, located in Beijing, China.

[0104] It should be noted that the above embodiments are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation of the present invention. Those skilled in the art can make other variations or modifications based on the above description. It is impossible to exhaustively list all possible implementations here. All obvious variations or modifications derived from the technical solutions of this invention are still within the scope of protection of this invention.

Claims

1. Vibrio parahaemolyticus ( Vibrio parahaemolyticus CJ5, characterized in that, The Vibrio parahaemolyticus CJ5 is deposited at the China General Microbiological Culture Collection Center (CGMCC) on June 26, 2025, with accession number CGMCC No. 34858, located in Beijing, China.

2. The application of Vibrio parahaemolyticus CJ5 as described in claim 1 in the degradation of sea cucumber fucoidan sulfate.

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