Kosakonia caroliniana strain bsx-h4 and application thereof

Abstract

The application discloses a Kosakonia cowanii BSX-h4 strain and application thereof. The Kosakonia cowanii BSX-h4 strain is preserved in the Guangdong Microbial Culture Collection Center, and the preservation date is January 8, 2024, and the preservation number is GDMCC No: 64253. The BSX-h4 strain can be used for fermenting coffee, so that the fermentation process of coffee is stabilized, the quality of coffee beans is improved, the production batch stability is improved, the types and contents of flavor substances such as alcohols, ketones, aldehydes, pyrazines and ethers in green coffee beans and roasted coffee beans are obviously improved, the fruit flavor of coffee beverages is improved, and the strain has food safety, so that the strain can be developed into a coffee flavoring microbial starter, and used for improving coffee flavor and improving coffee quality.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of coffee fermentation. More particularly, it relates to a Kosakonia cowanii BSX-h4 strain and its application. BACKGROUND

[0002] Coffee bean refers to the fruit of the beverage crop coffee of the genus Coffea in the Rubiaceae family. After peeling and fermentation, green coffee beans can be obtained, and roasted beans can be obtained after roasting, which can be used to make coffee drinks.

[0003] During the primary processing of green coffee beans, microbial fermentation can convert sugar substances in coffee fruits into aromatic precursor substances such as alcohols, acids, ketones, and aldehydes, thereby imparting unique flavors to green coffee beans. For example, furfuryl alcohol is the main source of coffee aroma, which has a pleasant roasting and smoking aroma; trans-nonenal imparts a fresh woody aroma to coffee; cycloalkenes and basic cyclopentanone impart a caramel aroma to coffee; maltol and 3-methylbutanal impart a sweet taste to coffee; furan derivatives can impart fruit flavors such as pineapple and mango to coffee. The flavor of commercially available roasted beans (commercial beans) is mainly affected by the volatile mixture formed during the roasting process of the aromatic precursor substances in the green beans, which is an extremely important sensory attribute of roasted beans, determining the price of roasted beans.

[0004] Currently, the primary processing of small-grain coffee in China mainly relies on traditional wet fermentation, which is greatly affected by the types of microorganisms and temperature, leading to inconsistent quality of coffee beans and poor production batch stability. Developing microbial fermentation agents and stabilizing the microbial composition in wet processing are crucial for the formation of coffee flavor, quality, and economic benefits. Although there are reports of fermentation strains that can reduce the bitterness of coffee, the types and contents of flavor substances such as alcohols, acids, and ketones in coffee beans fermented by different types of microorganisms differ significantly. Moreover, coffee flavors are diverse, and continuous exploration of fermentation strains that can enhance different coffee flavors is beneficial for the development of different flavor coffee products and the long-term development of the coffee industry. SUMMARY

[0005] The present application provides a Kosakonia cowanii strain to address the above technical problems. Fermentation of coffee beans using this strain can increase the content of flavor substances such as alcohols and esters in the resulting green coffee beans, thereby improving the flavor of coffee.

[0006] The first object of the present application is to provide a Kosakonia cowanii strain.

[0007] The second object of the present application is to provide a microbial agent containing the strain.

[0008] A third object of the present application is to provide the use of the strain or the microbial inoculum in fermenting coffee beans.

[0009] A fourth object of the present application is to provide the use of the strain or the microbial inoculum in preparing a preparation for fermenting coffee beans.

[0010] A fifth object of the present application is to provide the use of the strain or the microbial inoculum in improving the flavor of coffee.

[0011] A sixth object of the present application is to provide the use of the strain or the microbial inoculum in preparing a preparation for improving the flavor of coffee.

[0012] A seventh object of the present application is to provide a fermented coffee.

[0013] The above objects of the present application are achieved by the following technical solutions.

[0014] The present application has isolated and purified a strain of Kosakonia cowanii that can be used for fermenting coffee beans, named BSX-h4 strain. The use of the strain can not only stabilize the coffee fermentation process, improve the uneven quality of coffee beans, and the poor production batch stability problem, but also improve the types and contents of flavor substances such as alcohols, ketones, aldehydes, pyrazines, and ethers in the fermented coffee beans and roasted beans, enhance the fruity flavor of the coffee beverage, and improve the flavor of coffee. In addition, the fermented coffee beans and roasted beans obtained by using the strain do not contain sulfides. Therefore, the BSX-h4 strain and its applications are claimed in the present application.

[0015] Specifically, the Kosakonia cowanii BSX-h4 strain was deposited with the Guangdong Microbial Culture Collection Center on January 8, 2024, and the deposit number is GDMCC No: 64253.

[0016] The present application also provides a microbial inoculum containing the strain.

[0017] The use of the strain or the microbial inoculum in the present application can ferment coffee beans, stabilize the coffee fermentation process, and improve the content of flavor substances in the fermented coffee beans and the subsequent roasted beans. Therefore, the use of the strain or the microbial inoculum in fermenting coffee beans is claimed in the present application.

[0018] The present application also claims the use of the strain or the microbial inoculum in preparing a preparation for fermenting coffee beans.

[0019] The present application also claims the use of the strain or the microbial inoculum in improving the flavor of coffee.

[0020] The application also claims the use of the strain or the microbial inoculum of claim in the preparation of a preparation for improving the flavor of coffee.

[0021] Specifically, the improvement of the flavor of coffee is to increase the content of flavor substances in coffee beans.

[0022] Specifically, the increase of the content of flavor substances in coffee beans includes the increase of the content of substances that cannot be detected originally.

[0023] Specifically, the coffee beans include green coffee beans and roasted beans.

[0024] Specifically, the flavor substances include decanal, 5-methyl furfural, hexyl acetate, propyl hexanoate and / or furanone, which are characteristic aroma components that add fruity and floral characteristics to coffee. Among the flavor substances, decanal has a strong floral and fruity aroma, sometimes compared to the aroma of orange flowers or jasmine and the flavor of citrus fruits, and is widely used in the manufacture of perfumes and essences to provide floral and fruity flavors to products. 5-methyl furfural brings a sweet, caramel-like flavor characteristic of coffee. Hexyl acetate has a flavor similar to that of a ripe apple or pear. Propyl hexanoate has a flavor similar to that of an apple, pear or banana, with a slight fatty or cheesy aroma. Furanone has a rich fruity flavor and a jammy taste, and has a cranberry flavor when diluted.

[0025] Specifically, the coffee is Arabica coffee.

[0026] The application also provides a fermented coffee, which is prepared from coffee beans using the strain or the microbial inoculum of the application in the fermentation process.

[0027] Specifically, the content of flavor substances in the coffee is increased, and the flavor substances include decanal, 5-methyl furfural, hexyl acetate, propyl hexanoate and / or furanone.

[0028] The application has the following beneficial effects:

[0029] The application provides a Kosakonia cowanii strain, named BSX-h4 strain, which is preserved in the Guangdong Microbial Culture Collection Center on January 8, 2024, and has a preservation number of GDMCC No: 64253. The use of the BSX-h4 strain of the application for the fermentation of coffee not only stabilizes the coffee fermentation process and improves the uneven quality of coffee beans and the poor production batch stability, but also significantly increases the content of flavor substances such as alcohols, ketones, aldehydes, pyrazines and ethers in green coffee beans and roasted beans, enhances the fruity and floral flavors of the coffee beverage produced, and the strain has food safety, which can be developed as a coffee flavoring microbial starter for improving the flavor of coffee and improving the quality of coffee. BRIEF DESCRIPTION OF DRAWINGS

[0030] Figure 1 Figure a is the plate of BSX-h4 strain inoculated with streak; Figure b is the single colony of BSX-h4 strain.

[0031] Figure 2 Figure is the result of gram staining of BSX-h4 strain.

[0032] Figure 3 Figure is the phylogenetic tree constructed based on 16S rDNA sequence and gyrB sequence of BSX-h4 strain.

[0033] Figure 4 Figure is the statistical result of unique and common volatile substance species of green coffee beans of natural fermentation group (CK) and strain fermentation group (H4).

[0034] Figure 5 Figure is the statistical result of unique and common volatile substance species of roasted beans of natural fermentation group (CK) and strain fermentation group (H4). DETAILED DESCRIPTION

[0035] The present application will be further described in conjunction with the accompanying drawings and specific examples, but the examples do not limit the present application in any form. Unless otherwise specified, the reagents, methods and devices used in the present application are conventional reagents, methods and devices in the technical field.

[0036] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0037] The BSX-h4 strain described in the embodiments of the present application is Kosakonia cowanii BSX-h4 strain, which was preserved in Guangdong Microbial Culture Collection Center on January 8, 2024, with the preservation number of GDMCC No: 64253 and the preservation address of 5th Floor, Experimental Building, 100, Xianlie Middle Road, Yuexiu District, Guangzhou, Guangdong Province.

[0038] Example 1: Obtaining and identification of BSX-h4 strain

[0039] 1. Obtaining of BSX-h4 strain

[0040] The BSX-h4 strain described in the present application is obtained by using plate dilution method to separate and purify from the fermentation broth of Coffea arabica L., and the specific process is as follows:

[0041] The fermentation broth of Coffea arabica L. is filtered with six layers of gauze, and the filtrate is gradient diluted (10 -1 , 10 -2 , 10-3 , 10 -4 and 10 -5 ), respectively, 100 μL of different concentrations of the diluted filtrate was taken and coated in LB liquid medium, 3 repeats for each gradient; culture at 37°C, pick out single colonies and inoculate on LB solid medium; the separated strains were purified and cultured, and after 3 times of continuous purification, preserved by glycerol freezing method.

[0042] 2. Identification of BSX-h4 strain

[0043] (1) Morphological identification

[0044] The BSX-h4 strain was inoculated on LB solid medium and cultured at 37°C for 48 h, and then the morphology of single colonies was observed. The single colonies of BSX-h4 strain were inoculated in LB liquid medium and cultured at 37°C with 150 r / min for 16-24 h, and then a proper amount of bacterial solution was taken for Gram staining, and the Gram staining characteristics were observed under a microscope.

[0045] The colony morphology of BSX-h4 strain is shown in Figure 1 ; Figure 1 a in the figure is the plate inoculated with BSX-h4 strain; Figure 1 b in the figure is the single colony of BSX-h4 strain. As can be seen from Figure 1 , the colony of BSX-h4 strain is round, yellow, smooth-edged, wet-surfaced, opaque and convex in the middle.

[0046] The Gram staining result of BSX-h4 strain is shown in Figure 2 . As can be seen from Figure 2 , BSX-h4 strain is a Gram-negative bacterium, and the bacterial body is rod-shaped with a size of 0.5-0.8 μm x 4.0-6.0 μm.

[0047] (2) Molecular biological identification

[0048] The BSX-h4 strain was activated in LB solid medium and cultured at 37°C for 48 h, and then single colonies were picked and inoculated in LB liquid medium and cultured at 37°C with 150 r / min for 12 h; the bacterial body was collected by centrifugation, and the genomic DNA of BSX-h4 strain was extracted according to the instructions of OMEGA bacterial DNA extraction kit; the extracted genomic DNA of the strain was used as a template, and PCR amplification was performed with 16S rRNA primers and gyrB primers, respectively; the amplification products were sent to a sequencing company for sequencing after electrophoresis detection.

[0049] The nucleotide sequence of the 16S rRNA primer is as follows:

[0050] 27F: 5'-AGAGTTTGATCCTGGCTCAG-3';

[0051] 1492R: 5'-GGTTACCTTGTTACGACTT-3'.

[0052] The nucleotide sequences of the gyrB primers are as follows:

[0053] F: 5'-TTGGTGCTTCTGTTGTT-3';

[0054] R: 5'-CATCCCCAAAGCCAG-3'.

[0055] The PCR reaction system (total 25 μL) is as follows: 2x PCR Buffer 12.5 μL, 2 mM dNTPs 5 μL, 10 pmoL / mL upstream primer 0.75 μL, 10 pmoL / mL downstream primer 0.75 μL, KOD FX (1.0 U / μL) 0.5 μL, DNA 1.0 μL, ddH2O to 25 μL. The PCR amplification procedure is as follows: 94°C pre-denaturation for 5 min, 98°C denaturation for 10 s, 55°C annealing for 30 s, 72°C extension for 30 s, a total of 35 cycles, and finally 72°C extension for 5 min.

[0056] The 16S rDNA sequence of the BSX-h4 strain is as shown in SEQ ID NO. 1, and the gyrB sequence is as shown in SEQ ID NO. 2.

[0057] BLAST comparison and analysis of the sequencing results of the 16S rDNA sequence and the gyrB sequence of the BSX-h4 strain in the NCBI database with the 16S rDNA and gyrB sequences of existing strains found that the BSX-h4 strain has a similarity of more than 98% with the 16S rDNA sequence and the gyrB sequence of multiple Kosakonia cowanii. Based on the 16S rDNA sequence and the gyrB sequence of the BSX-h4 strain, a phylogenetic tree was constructed using the MEGA 11 software by the Neighbour-joining method, as shown in FIG. 1. It can be known from FIG. 1 that the BSX-h4 strain has the closest genetic relationship with the reported Kosakonia cowanii. Figure 3 Figure 3 Based on the morphological and molecular biological identification results of the BSX-h4 strain, the BSX-h4 strain was identified as Kosakonia cowanii.

[0058] Example 2 Detection of the edible safety of the BSX-h4 strain

[0059] ​The edible safety of the BSX-h4 strain was detected according to GB 15193.3-2014 Food Safety National Standard Acute Oral Toxicity Test. ICR mice with a body weight range of 18-22 g were selected, 20 males and 20 females, and were fed in 8 cages, 5 females or 5 males per cage. After 2 days of adaptation in the ABSL-2 feeding area of the Yunnan University Experimental Animal Center (20-26°C, relative humidity 40%-70%), the gavage test was performed. The mice were fasted for 4 h before the test, and the mice were required to continue fasting for 1 h after being given the BSX-h4 bacterial suspension. The mice were allowed to drink water during the fasting period. The BSX-h4 strain (OD 600 = 3.0) was resuspended for use after being washed twice with sterile water. The mice were gavaged at doses of 0, 5, 10, and 15 g / kg, and the gavage volume was 30 mL / kg. All mice were observed for 7 days after gavage, and the number of dead mice was recorded.

[0060] The results of the edible safety test of the BSX-h4 strain are shown in Table 1. As shown in Table 1, no mice died, and no poisoning occurred during the 7-day observation after gavage of the mice, and the mice were active during the test period, indicating that the BSX-h4 strain is safe and non-toxic.

[0061] Table 1 Death of mice at different gavage doses

[0062]

[0063] Example 3 Effect of fermentation treatment of BSX-h4 strain on the volatile substance content of green coffee beans and roasted beans

[0064] 1. Fermentation and roasting treatment of coffee beans

[0065] The fermentation treatment of coffee beans was divided into two groups, namely a natural fermentation group (CK) and a strain fermentation group (H4). Both groups of fermentation treatment systems retained the microorganisms naturally carried by coffee beans, and the systems contained microorganism strains from natural sources such as fruit peels, fruit flesh, and picking environments. The difference between the strain fermentation group and the natural fermentation group was that the strain fermentation group was artificially inoculated with 1% (v / v) of BSX-h4 strain active liquid (bacterial concentration was 5.0 log CFU / mL) in the fermentation system.

[0066] The specific process was as follows: coffee fresh fruits collected at the same site were mechanically peeled, and 2.0 kg of peeled coffee beans were placed in 8.0 L glass fermentation tanks, and 4.0 L of deionized water was added. The H4 fermentation group was added with 1% (v / v) of BSX-h4 strain active liquid, and the control group was added with an equal amount of deionized water. Each group of coffee bean fermentation systems was placed in a 30°C constant temperature incubator for fermentation for 60 h. Three replicates were set for each group.

[0067] After the fermentation treatment, the coffee beans in each group are placed in a cool environment for natural drying, and a coffee shelling machine is used to remove the coffee bean shells and silver skins to obtain green coffee beans, which are stored in a sealed environment at room temperature; the green coffee beans obtained after the fermentation treatment are taken and subjected to roasting treatment under the same conditions. The roasting conditions are as follows: 20 g of coffee beans are roasted each time, a coffee bean roaster is used for roasting, the temperature of the coffee beans is 160℃, dehydration starts when the temperature rises to 180℃, after 10 minutes of dehydration, the temperature is adjusted to the roasting temperature of 220℃ for roasting, and the roasting time is 10 minutes.

[0068] 2. Detection of volatile substances in green coffee beans and roasted beans

[0069] The present application uses gas chromatography-mass spectrometry (GC-MS) technology to detect and analyze the volatile substances contained in the green coffee beans and roasted beans of the natural fermentation group and the strain fermentation group.

[0070] The specific process is as follows: the green coffee beans and roasted beans obtained after the fermentation treatment in each group are ground into powder and passed through a 20-mesh sieve for use; 6 g of the prepared green coffee bean and roasted bean powder is accurately weighed in a 100 mL blue cap bottle, 50 mL of ethyl acetate (boiling point 76.5-77.5℃) is added, and the mixture is placed in an ultrasonic constant temperature water bath oscillator for extraction for 1 hour, then it is left to stand for 20-30 minutes to separate the layers, the upper liquid is sucked and filtered twice with qualitative filter paper, the filtrate is transferred to a rotary bottle and evaporated with a rotary evaporator to completely remove the ethyl acetate, the rotary bottle is removed after no liquid drops are dropped, 2 mL of chromatographic grade dichloromethane is added and mixed with the coffee oil in the bottle, 1 mL of a syringe is used to filter the mixture through a 0.22 μm needle filter membrane into a sample bottle, and the sample bottle is placed in a GC-MS instrument for detection of volatile substances.

[0071] Microextraction conditions: solid phase microextraction head (SUPELCO): 50 / 30 μm PDMS / DVB / CAR, Stableflex; aging temperature of extraction head: 250℃, aging time: 300s, insertion depth: 20mm, coating extension length: 12mm; the sample is preheated in a 20 mL headspace bottle, heating temperature: 40℃, extraction temperature: 60℃, extraction time: 2400s, insertion depth: 15mm, coating extension length: 12mm, stirring speed: 300r / min, stirring time: 600s, analysis temperature: 270℃, analysis time: 300s, insertion depth: 20mm, coating extension length: 12mm.

[0072] Gas chromatography conditions: chromatographic column: Agilent 19091s-433HP-5MS (30m x 0.25mm x 0.25um); column temperature: using programmed temperature, 40℃ for 5min, then 2℃ / min to 70℃, 2min, 3℃ / min to 120℃, 5℃ / min to 50℃, finally 10℃ / min to 230℃, 2min, flow rate is 1.4mL / min. Detector: MSD detector; carrier gas He (purity 99.999%), 40mL / min; split injection: split ratio 5:1; septum purge flow is 3mL / min; injection port temperature: 270℃.

[0073] Mass spectrometry conditions: ion source: EI source, electron energy 70eV; ion source temperature: 230℃; quadrupole temperature is 150℃; transfer line temperature: 280℃; acquisition mode: scan mode Scan / SIM; scan mass range: 35-550; solvent delay 1.0min.

[0074] 3, Analysis results of volatile substances

[0075] (1) Analysis results of volatile substances of coffee beans

[0076] The volatile substances of the two groups of coffee beans treated by fermentation were analyzed in combination with the spectrum library and NIST spectrum library of GC-MS, 68 kinds of compounds were detected in the samples of CK and H4 groups, which were divided into 9 categories of alcohol, ester, acid, ketone, aldehyde, pyrazine, furan, pyrrole and other substances, as shown in Table 2. As shown in Table 2, after inoculating BSX-h4 strain for fermentation, the relative contents of alcohol, ester, acid, ketone, aldehyde, pyrazine, furan, pyrrole and other substances in the obtained coffee beans are increased. Based on the analysis results of GC-MS, the unique and common volatile substance categories of the coffee beans of CK and H4 groups were counted, and the results are shown in Table 3. Figure 4 As shown in Table 3, from the specific component categories, 41 and 53 kinds of substances were detected in CK and H4 groups respectively, of which 15 kinds were unique to CK, 27 kinds were unique to H4, and 26 kinds of substances were common to both. Figure 4

[0077] Table 2 Analysis of volatile substances in coffee beans of CK and H4 groups

[0078]

[0079]

[0080]

[0081] nd in the table indicates not detected. ​

[0082] (2) Volatile substances analysis results of roasted beans

[0083] The same as the foregoing, the volatile substances of the two groups of roasted beans after fermentation and roasting treatment were analyzed in combination with the spectrum library and NIST spectrum library of GC-MS, and the volatile substances detected in the CK and H4 groups of samples are specifically shown in Table 3. As can be seen from Table 3, 104 kinds of volatile compounds were identified in the 2 groups of samples, including 11 kinds of substances such as alcohols, esters, ethers, acids, phenols, ketones, aldehydes, pyrazines, pyrroles, furans and other substances. Compared with the natural fermentation group, the number of alcohols, phenols, ketones, aldehydes, pyrroles and pyrazines in the inoculation BSX-h4 strain fermentation group increased, and ether substances were also newly added. Based on the GC-MS analysis results, the unique and common volatile substances of the CK and H4 groups of roasted beans were counted, and the results are shown in Table 4. Figure 5 As can be seen from the specific component types, 57 and 75 kinds of substances were detected in the CK and H4 groups, respectively, of which 28 kinds were unique to CK, 46 kinds were unique to H4, and 29 kinds of substances were common to both Figure 5 ).

[0084] Table 3 Component analysis of volatile substances in roasted beans of CK and H4 groups

[0085]

[0086]

[0087]

[0088]

[0089] nd in the table represents not detected.

[0090] It was detected that the component analysis results of volatile substances of each repeat group were not much different, indicating that the BSX-h4 strain described in the present application can be used for fermentation treatment of coffee, which can stabilize the coffee fermentation process, and improve the problems of uneven quality of coffee beans and poor production batch stability.

[0091] Example 4 Coffee characteristic aroma evaluation

[0092] In addition to detecting and analyzing volatile substances in coffee green beans and roasted beans, the present application also refers to the SCAA cupping system to cup the roasted beans of the natural fermentation group and the strain fermentation group respectively, and evaluate the coffee characteristic aroma. The specific process is: accurately weigh 8.50 g of freshly ground coffee powder in the cupping, pour 150 mL of 93°C hot water, and form an evaluation group of 4 Q-Grader international coffee quality tasters, score 10 indexes of coffee dry and wet aroma, flavor, aftertaste value, acidity, fullness, balance, consistency, cleanliness, sweetness and overall evaluation, among which the consistency, cleanliness and sweetness indexes are given full marks. The sum of the scores of all 10 sensory indexes is the final score of a single sample, and the brewed coffee liquid is described in terms of sensory smell.

[0093] The results of coffee characteristic aroma evaluation of roasted beans in CK and H4 groups are shown in Table 4. As shown in Table 4, the total scores of roasted beans before and after inoculation are 81.5 and 82.5 respectively; among them, the roasted beans after inoculation have improved in aftertaste value, acidity, fullness and balance compared with the roasted beans before inoculation.

[0094] Table 4 Coffee characteristic aroma evaluation results of roasted beans in CK and H4 groups

[0095]

[0096] The coffee characteristic aroma sensory description of roasted beans in CK and H4 groups is shown in Table 5.

[0097] Table 5 Coffee characteristic aroma sensory description of roasted beans in CK and H4 groups

[0098]

[0099] From Table 5, it can be seen that the fruit aroma of the inoculated fermentation group is more abundant than that of the natural fermentation group, and in addition, the flower aroma is also added. In combination with the volatile substance analysis results of Example 3, the fruit aroma added after the inoculation of the roasted beans is related to the ester substance. Hexyl acetate is usually described as a fruit flavor, especially a flavor similar to a ripe apple or pear. Propyl hexanoate also has a special fruit aroma, especially a flavor similar to an apple, a pear or a banana and has a slight fat or cheese aroma. The relatively higher content of propyl hexanoate in the H4 group of roasted beans is one of the reasons why the fruit aroma of the H4 roasted beans is more abundant. In addition, furanone has a rich fruit aroma and a jam flavor, and after dilution, it has a cassis flavor. The newly added flower aroma in the H4 group of roasted beans is related to decanal, which has a strong floral and fruity aroma, and is sometimes compared to the aroma of orange flowers or jasmine and the aroma of citrus fruits, especially oranges and lemons. Decanal also exists in many plants in nature, especially in the essential oils of flowers and citrus fruits, and is therefore widely used in the manufacture of perfumes and essences to provide floral and fruity aromas to products. The 5-methyl furfural in the aldehyde substance brings a sweet aroma, a caramel-like flavor characteristic of coffee. Most of the ketone substances are nutty and caramel-flavored. The newly added ketone substance in the inoculated fermentation group makes the coffee have a coffee alcohol aroma while mixing with the aroma of fruits, making the main aroma of coffee more refreshing and balanced.

[0100] The above examples are the preferred embodiments of the present application, but the embodiments of the present application are not limited to the above examples, and any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present application should be equivalent replacement methods, and are all included in the protection scope of the present application.

Claims

1. A strain of Coxella sacchariformis ( Kosakonia cowanii BSX-h4 strain, characterized in that, The strain was deposited in the Guangdong Microbial Culture Collection Center on January 8, 2024, and the deposit number is GDMCC No: 64253.

2. An inoculant containing the BSX-h4 strain of claim 1.

3. Use of the BSX-h4 strain of claim 1 or the inoculant of claim 2 in fermenting coffee beans.

4. Use of the BSX-h4 strain of claim 1 or the inoculant of claim 2 in the preparation of a preparation for fermenting coffee beans.

5. Use of the BSX-h4 strain of claim 1 or the inoculant of claim 2 in improving the flavor of coffee.

6. Use according to claim 5, characterized in that, The improvement of the flavor of coffee is to increase the content of flavor substances in coffee beans.

7. Use according to claim 6, characterized in that, The flavor substances include decanal, 5-methylfurfural, hexyl acetate, propyl hexanoate and / or furanone.

8. Use of the BSX-h4 strain of claim 1 or the inoculant of claim 2 in the preparation of a preparation for improving the flavor of coffee.

9. Use according to claim 8, characterized in that, The improvement of the flavor of coffee is to increase the content of flavor substances in coffee beans.

10. Use according to claim 9, characterized in that, The flavor substances include decanal, 5-methylfurfural, hexyl acetate, propyl hexanoate and / or furanone.

11. A fermented coffee, characterized in that, The coffee is prepared from coffee beans using the BSX-h4 strain of claim 1 or the inoculant of claim 2 in the fermentation process.

12. The coffee according to claim 11, characterized in that, The content of flavor substances in the coffee is increased, and the flavor substances include decanal, 5-methylfurfural, hexyl acetate, propyl hexanoate and / or furanone.

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