A strain of candida famata and its use in tobacco products

Abstract

The application belongs to the field of agricultural microorganism technology, and discloses a strain of producing aroma heterologous Candida and its application in tobacco products. The strain can meet the needs of different tobacco products, has excellent stress resistance, and the fermentation liquor of the strain can produce 54 kinds of characteristic aroma substances with high content, the propylene glycol extract can produce 52 kinds of characteristic aroma substances. After the fermentation treatment of flue-cured tobacco leaves, compared with the untreated tobacco leaves, the aroma quality and amount increase, the irritancy decreases, the sweetness increases, the comfort improves, the fermented cigar leaves after airing can reach the product smoking standard after 30 days of intensified fermentation, the agricultural fermentation time is shortened, and after the fermentation is finished, the honey sweet aroma, clean sweet aroma, floral aroma and other aroma of the cigar leaves are improved, and the cigar style is highlighted. The heterologous Candida in the application is an aroma-producing strain, can directionally improve the flavor quality of tobacco products, and has a good application prospect in aroma enhancement and quality improvement of products such as cigarettes and cigars.

Description

Technical Field

[0001] This invention belongs to the field of agricultural microbial technology, specifically relating to an aroma-producing heterologous Candida yeast and its application in tobacco products. Background Technology

[0002] As a global economic crop, tobacco products are evolving towards reduced harm, richer flavor, and enhanced sensory comfort. Tobacco is the primary raw material for cigarettes. Cigarettes, with their unique aroma characteristics and convenient consumption, have become the core product for the traditional tobacco industry to meet market demands. However, with the increasing demand for high-quality cigarettes, higher requirements are being placed on cigarette quality improvement and aroma enhancement technologies. Research shows that the formation of characteristic cigarette flavors is closely related to the metabolic regulation of the microbial community during tobacco raw material fermentation, playing an irreplaceable role in the transformation of aroma components.

[0003] Microbial fermentation has great potential in enhancing the aroma and quality of cigarettes. It mainly involves directly inoculating aroma-producing microorganisms onto the surface of tobacco leaves for fermentation or using these microorganisms to prepare tobacco flavorings and adding them to tobacco for aroma enhancement. This method is not limited by climate, has a short production cycle, and produces high-quality, diverse aroma compounds. Currently, the main microorganisms used in tobacco leaves and cigarette products are Clostridium species (…). Clostridium ), Micrococcus ( Micrococcus ), Pseudomonas ( Peudomonas putida ), Bacillus ( Bacillu ) and brewer's yeast ( Saccharomyces cerevisiae Research on the application of Candida albicans in improving the flavor and quality of tobacco is relatively limited; its application is more prevalent in fermented foods (soy sauce, fermented soybean paste, fermented black beans, etc.). For example, existing reports indicate that Candida albicans SY152 is used in the fermentation of fermented soybean paste or soy sauce to enhance the aroma and significantly improve the taste and flavor; Candida esculenta is used to enhance the fermentation function of low-salt raw fermented soybean paste, improving its color, flavor, and typicality; and Candida tropicalis JSY83 is used in the production of Daqu (a type of starter culture) and strong-aroma baijiu (Chinese liquor) to increase the yield of high-quality products and the overall alcohol yield.

[0004] Therefore, there are currently no reports on the application of aroma-producing Candida heterotrophicum in tobacco products. Summary of the Invention

[0005] To address the aforementioned issues, and considering the lack of reports on the preparation of flavorings and fragrances to enhance flavor quality based on aroma-producing Candida albicans in the production of cigarettes and other tobacco products, this invention provides an aroma-producing strain of Candida albicans (…). Candida allociferrii ) strains and their application in tobacco products.

[0006] In a first aspect, the present application provides a xanthogenic alien Candida, which is classified as alien Candida Candida allociferrii TC5, with the accession number CCTCC NO: M 20241402, which was deposited at the China Center for Type Culture Collection on July 1, 2024, and the address of the depositary is Wuhan University, Wuhan, China.

[0007] In the present application, the monoclonal colony of the alien Candida has the following morphological characteristics: white, round, raised, smooth surface, moist, and also has the fermentation characteristics of high temperature resistance, high acid resistance, high nicotine resistance, and directional xanthogenicity. The alien Candida has a good temperature tolerance range, and the strain still has good growth conditions in the range of 15-45℃, which can meet the needs of the entire fermentation production stage of cigar tobacco.

[0008] In a second aspect, the present application further provides a composition for improving the flavor quality of tobacco products, wherein the main active ingredient of the composition comprises the alien Candida biological inoculant, the fermentation broth of the alien Candida, or the propylene glycol extract of the alien Candida.

[0009] In an embodiment of the present application, the alien Candida biological inoculant further comprises Bacillus, yeast, and / or white rot fungus.

[0010] In a third aspect, the present application further provides the use of the xanthogenic alien Candida of the first aspect of the present application or the composition of the second aspect of the present application in the preparation of tobacco products.

[0011] In an embodiment of the present application, the tobacco product is a tobacco flavor, tobacco extract, tobacco sheet, flue-cured tobacco, electronic cigarette liquid, or cigar tobacco, etc.

[0012] In an embodiment of the present application, the tobacco product is a tobacco flavor, which is prepared using the fermentation broth of the alien Candida or the propylene glycol extract of the alien Candida as raw materials.

[0013] In an embodiment of the present application, the tobacco product is a tobacco extract, which is obtained by fermenting tobacco extract using the alien Candida.

[0014] In an embodiment of the present application, the tobacco product is a reconstituted tobacco sheet, which is prepared by fermenting stem paste using the alien Candida and then using the papermaking method.

[0015] In an embodiment of the present application, the tobacco product is an electronic cigarette liquid, which is prepared using the propylene glycol extract of the alien Candida strain.

[0016] In an embodiment of the present application, the tobacco product is cigarette tobacco leaf, and the cigarette tobacco leaf is prepared by using the Candida utilis liquid directly acting on the cigarette tobacco leaf or spraying the Candida utilis liquid as a tobacco leaf aging agent on the surface of tobacco leaf to be aged for accelerating tobacco aging or applying to the cigarette leaf in the threshing and redrying stage or spraying on the cut tobacco in the cut tobacco stage.

[0017] In an embodiment of the present application, the tobacco product is cigar tobacco leaf, and the cigar tobacco leaf is prepared by using the Candida utilis liquid directly acting on or spraying the Candida utilis liquid as a reinforced fermentation agent on the surface of the cigar tobacco leaf to be fermented or adding the Candida utilis liquid to the surface of the cigar tobacco leaf to be fermented by a pressurized atomization method for reinforced fermentation.

[0018] In a fourth aspect, the present application further provides an application of the aroma-producing Candida utilis in the first aspect or the composition in the second aspect to the directional production of a plurality of odor substances including floral, mushroom, cocoa, rose, almond and the like.

[0019] In an embodiment of the present application, the odor substances include the following substances,

[0020] esters: methyl benzoate, methyl anthranilate,

[0021] alcohols: 1-octen-3-ol, 3-methyl-1-butanol, 2-ethylhexanol and the like,

[0022] aldehydes: benzaldehyde, nonanal,

[0023] ketones: isophorone, acetophenone, geranyl acetone,

[0024] phenols: 2-methoxy-4-vinylphenol.

[0025] The present application has the following technical effects relative to the prior art:

[0026] (1) The aroma-producing Candida utilis TC5 provided by the present application can produce 54 characteristic aroma substances in the fermentation broth of the culture medium, and the directional high-yield aroma substances include esters (methyl benzoate, methyl anthranilate and the like) and ketones (acetophenone, geranyl acetone) with floral and fruity aroma, and alcohols (1-octen-3-ol, 3-methyl-1-butanol, 2-ethylhexanol and the like) with mushroom, cocoa and rose aroma.

[0027] (2) When the aroma-producing Candida utilis TC5 provided by the present application is used for reinforced fermentation of flue-cured tobacco leaf, the aroma quality and aroma amount are increased, the irritation is reduced, the sweetness is increased, and the comfort is improved compared with the untreated tobacco leaf.

[0028] (3) When using the aroma-producing heterologous Candida TC5 provided by the present invention for the production of cigar enhanced fermentation, compared with normal fermentation, the fermentation time of 30 days can reach the smoking standard, shortening the aging time. After fermentation, the aroma of cigar tobacco leaves such as honey sweetness, light sweetness, and floral aroma is enhanced, the cigar style is highlighted, the richness and aftertaste are improved, and the sensory quality of smoking is significantly higher than that of non-enhanced fermented tobacco leaves. The main chemical components meet the national standard requirements for cigars. Attached Figure Description

[0029] Figure 1 The aroma-producing heterologous Candida albicans of the present invention ( Candida allociferrii Colony morphology and microscopic morphology of TC5; where A is a plate colony image and B is a microscopic observation image of the strain.

[0030] Figure 2 The aroma-producing heterologous Candida albicans of the present invention ( Candida allociferrii Phylogenetic tree of TC5 strain.

[0031] Figure 3 The aroma-producing heterologous Candida albicans of the present invention ( Candida allociferrii Sensory evaluation radar charts of the fermentation broth and propylene glycol extract of TC5 strain in cigarettes; where A represents the functional improvement effect on cigarettes and B represents the aroma improvement effect on cigarettes. Detailed Implementation

[0032] The specific embodiments of the present invention are described below with reference to the accompanying drawings. Unless otherwise specified, the experimental methods used in the embodiments are conventional methods; the materials and reagents used are commercially available unless otherwise specified.

[0033] The main chemical components (total sugar, reducing sugar, total nitrogen, total alkaloids, potassium, and chlorine) of flue-cured tobacco and cigar tobacco leaves were detected using a continuous flow analyzer. The aroma-producing Candida albicans fermentation broth and volatile flavor compounds of tobacco leaves were detected using gas chromatography-mass spectrometry (GC-MS). Sensory evaluation of cigarettes and cigars was conducted in accordance with tobacco industry standards and relevant industrial enterprise standards.

[0034] PBS buffer: Weigh 8.0 g sodium chloride, 0.2 g potassium chloride, 1.44 g disodium hydrogen phosphate, and 0.24 g potassium dihydrogen phosphate, dissolve them in 800 mL distilled water, adjust the solution to 7.4 with HCl, and finally add distilled water to make up to 1 L to obtain 0.01 M PBS buffer.

[0035] PDA medium: Potato extract powder 12.0 g / L, glucose 20.0 g / L, agar 14.0 g / L, dissolved in distilled water by heating, dispensed, and autoclaved at 121℃ for 15 min before use.

[0036] Saccharification liquid culture medium: mix raw rice and water (1:4 mass ratio), add 2‰ of liquefying enzyme and 1‰ of saccharifying enzyme based on the mass of raw rice, 60℃ water bath, saccharify and liquefy for 4-6 hours, adjust the sugar content to 13 Brix, sterilize at 115℃ for 20 minutes, and then cool down for standby.

[0037] Cigar leaf culture medium preparation: 3.0 g of cigar leaf is cut into small pieces, 50 mL of distilled water is added, and oscillation is performed at 200 rpm for 30-60 minutes. After sterilization at 121℃ for 20 minutes, the sterilized cigar leaf culture medium is obtained.

[0038] Example 1: Screening and identification of strains

[0039] (1) Sample pretreatment

[0040] 5 g of fermented cigar leaf or alcoholized flue-cured tobacco leaf sample is weighed, cut into small pieces, and added to 50 mL of 0.85% sterile physiological saline. The sample is placed in a stacked temperature oscillation incubator, set to a temperature of 30℃ and a rotation speed of 150 r / min, and shaken for 2 hours.

[0041] (2) Dilution and plate coating

[0042] The supernatant of the sample in step 1 is diluted with sterile water in stages (10 -1 ~ 10 -8 ), and different concentrations of the diluents are coated on solid culture medium to allow the growth of single colonies of microorganisms on the medium. PDA solid culture medium is used for culture, and then placed in a constant temperature and humidity incubator for 48 hours of culture.

[0043] (3) Three-zone streaking separation

[0044] According to the morphological differences, different single colony strains in step (2) are selected and three-zone streaked on new PDA plates. The streaked plates are inverted and cultured in a constant temperature and humidity incubator at 30℃ for 48 hours. After multiple purification, multiple strains with aroma-producing effect are obtained. The preliminarily screened aroma-producing strains are inoculated in PDA liquid culture medium for enrichment culture for 48 hours, and the best aroma-producing strain is selected by smell.

[0045] (4) Purification and preservation

[0046] Step (3) is repeated for 2-3 times of purification until only single morphological characteristic single colony is obtained on the plate. Then, the single colony is inoculated in liquid PDA culture medium, placed in a shaker at 30℃ and 150 r / min for 48 hours of culture. Then, the bacterial solution is mixed with glycerol at a ratio of 1:1, and then preserved in a 2 mL bacteria preservation tube at -80℃ after numbering.

[0047] (5) Strain identification

[0048] The strain preserved in step (4) was selected, inoculated on PDA solid medium, and placed in a 30°C constant temperature and humidity incubator for 48 h. The colony morphology was observed, and the cell morphology was observed under a biological microscope after crystal violet staining. As shown in FIG. 1, the monoclonal colony had the following characteristics: white, round, raised, smooth surface, moist, and long silk-like pseudohyphae under a microscope. A single colony on the plate was taken in sterile water, boiled to extract DNA, amplified, and sequenced. Through ITS rDNA sequencing, the nucleotide sequence was SEQ ID NO. 1, and the NCBI database comparison analysis showed that the homology of the strain with Candida heterohypha was 100%, and the strain could be identified as Candida heterohypha. The strain was preserved in the China Center for Type Culture Collection on July 1, 2024, with the preservation number CCTCC NO: M 20241402, and the preservation address was Wuhan University, Wuhan, China. Figure 1 Figure 2

[0049] (6) Strain tolerance analysis

[0050] The pH, temperature, and nicotine tolerance of the strain TC5 were analyzed, and the results showed that the strain could grow at 15°C-45°C, pH 4-11, and nicotine content of 8000 mg / L or less.

[0051] Example 2 Determination of volatile flavor compounds in the fermentation broth of the Candida heterohypha strain TC5

[0052] (1) Preparation of TC5 fermentation broth

[0053] The Candida heterohypha preserved in the glycerol tube was streaked to obtain single colonies. The purified TC5 was inoculated into 50 mL of saccharification liquid at an inoculation amount of 2% (volume fraction), centrifuged to obtain the supernatant, and poured into a brown glass bottle to obtain a Candida heterohypha fermentation broth with floral and sweet aroma.

[0054] (2) Determination of volatile flavor substances

[0055] The volatile flavor substances in the fermentation broth of the Candida heterohypha strain were determined by headspace solid-phase microextraction-gas chromatography-mass spectrometry. 2.0 g of sodium chloride was added to a 20 mL headspace bottle, 6 mL of the supernatant obtained by centrifugation was placed in a 20 mL headspace bottle, 2 μL of 2-octanol (50 ppm) internal standard was added, and the sample was detected after extraction.

[0056] (3) Analysis of volatile flavor substances

[0057] ​​The NIST14 database was used to qualitatively analyze the volatile flavor substances in the fermentation liquor to determine the volatile flavor compound categories, and semi-quantitative analysis was performed based on the internal standard to determine the relative content of the main flavor compounds. The results are shown in Table 1.

[0058] Table 1 Volatile flavor substances in the fermentation liquor of TC5

[0059]

[0060]

[0061]

[0062] Example 3 Determination of volatile flavor compounds in the propylene glycol extract of the heterologous Candida strain TC5

[0063] (1) Preparation of propylene glycol solvent extract

[0064] The heterologous Candida strain preserved in glycerol was streaked to obtain a single colony. After the TC5 strain was inoculated and spread on a PDA plate for culture, the colonies on the plate had a strong floral fragrance. The strain was gently scraped off (2.0 g) with an inoculation loop and mixed with 20 mL of propylene glycol. After standing for 12 h, the supernatant was obtained by centrifugation and poured into a brown glass bottle to obtain the propylene glycol extract of the heterologous Candida strain.

[0065] (2) Determination of volatile flavor substances

[0066] The volatile flavor substances in the propylene glycol extract of the heterologous Candida strain were determined by headspace solid-phase microextraction-gas chromatography-mass spectrometry, as described in Example 2(2).

[0067] (3) Analysis of volatile flavor substances

[0068] The NIST14 database was used to qualitatively analyze the volatile flavor substances in the propylene glycol extract to determine the volatile flavor compound categories, and semi-quantitative analysis was performed based on the internal standard to determine the relative content of the main flavor compounds. The results are shown in Table 2.

[0069] Table 2 Volatile flavor substances in the propylene glycol solvent extract

[0070]

[0071]

[0072]

[0073] Example 4 Determination of volatile flavor compounds in the fermented tobacco dust of the heterologous Candida strain TC5

[0074] (1) Preparation of TC5 fermentation broth

[0075] The heterologous Candida glycerol-preserved was streaked to obtain a single clone, and the purified TC5 was inoculated into 50 mL of the heterologous Candida tobacco culture medium at an inoculation amount of 2% (volume fraction), and was subjected to 150 r·min -1 -36 h at 30℃ on a shaking table, centrifuged at 4000 r·min -1 for 10 min, and the supernatant was taken and poured into a brown glass bottle to obtain the heterologous Candida tobacco culture medium culture solution.

[0076] (2) Determination of volatile flavor substances

[0077] The volatile flavor substances in the heterologous Candida tobacco culture medium culture solution were determined by headspace solid-phase microextraction-gas chromatography-mass spectrometry, and the specific operation was the same as that in Example 2(2).

[0078] (3) Analysis of volatile flavor substances

[0079] The volatile flavor substances in the tobacco culture medium culture solution were qualitatively analyzed by using the NIST14 database to determine the volatile flavor compound categories, and the relative contents of the main flavor compounds were semi-quantitatively analyzed based on internal standards, and the results are shown in Table 3.

[0080] Table 3 Volatile flavor substances in the tobacco culture medium culture solution

[0081]

[0082]

[0083]

[0084]

[0085] Example 5 Preparation of tobacco flavor by using heterologous Candida TC5

[0086] The purified TC5 was inoculated into 50 mL of the saccharified liquid at an inoculation amount of 2%, and was subjected to fermentation for 36 h, and the supernatant obtained by centrifugation was the fermentation broth; the TC5 strain was inoculated and coated on a PDA plate, and was cultured at 30℃ for 48 h, and then the strain was gently scraped off (2.0 g) using an inoculation loop and was fully mixed with 20 mL of propylene glycol, and after standing for 12 h, the supernatant obtained by centrifugation was the propylene glycol extract of the bacterial cells.

[0087] The sensory evaluation results of the cigarettes injected with the fermentation broth and the propylene glycol extract are shown in Table 4. Figure 3As shown in the figure, the sensory quality indicators of the injected cigarettes changed to varying degrees compared to the control. In terms of aroma characteristics, both effectively improved the aroma quality, quantity, concentration, and permeability of the cigarettes, while reducing off-flavors. Regarding taste characteristics, both produced a smooth and delicate smoke. However, the fermentation liquid slightly increased the irritation and residue of the smoke, and the propylene glycol extract also showed slight residue, possibly due to the presence of small amounts of microbial metabolites such as proteins in the extract. Except for the irritation and residue indicators, the fermentation liquid was more effective than the propylene glycol extract in improving sensory characteristics. The fermentation liquid exhibited a roasted and sweet aroma, while the propylene glycol extract displayed a fresh, fruity, woody, floral, and sweet aroma, with sweetness being the most prominent. The results indicate that both the fermentation liquid and the propylene glycol extract can improve the aroma and taste characteristics of cigarettes and enhance their aroma and flavor.

[0088] Example 6: Preparation of tobacco extract from Candida heterologa TC5

[0089] Following the method described in step 1 of Example 2, the aroma-producing Candida heterotrophic yeast fermentation broth was obtained in advance. After centrifugation, the yeast cells were redissolved and mixed with sterile water. 200 g of commercially available tobacco extract was selected as raw material and placed in a 500 mL Erlenmeyer flask. The Candida heterotrophic yeast fermentation broth was inoculated at a ratio of 1%-10%, and then fermented at 30℃ in a constant temperature and humidity incubator for 3-5 days. After fermentation, the tobacco extract obtained after fermentation was dissolved and diluted with sterile water, and then sprayed onto unflavored tobacco at an addition rate of 0.05-0.1%. The flavor quality was evaluated by professional judges according to the national standard GB 5606.4-2005 "Sensory Technical Requirements for Cigarettes" and the sensory evaluation method for tobacco additives. The results showed that the aroma-producing Candida heterotrophic yeast strain could significantly improve the quality of tobacco extract through fermentation, increasing aroma, reducing off-flavors, reducing irritation, improving comfort, and increasing sweetness.

[0090] Example 7 Application of aroma-producing heterologous Candida TC5 in tobacco sheets

[0091] Following step 1 in Example 2, the aroma-producing Candida albicans fermentation broth was obtained in advance. After centrifugation, the cells were redissolved and mixed with sterile water, resulting in a bacterial concentration of 10. 6 -10 8CFU / mL, the bacterial solution was inoculated into the stem extract at a ratio of 5%-10% (dry matter mass fraction) according to the moisture content of the stem extract. Fermentation was then carried out at 30℃ for 1-12 h. The resulting product was coated onto a tobacco sheet substrate at a coating rate of 20%-50%, dried at 90℃ for 10 min, and then rehydrated to a moisture content of 10-13% before being shredded. The tobacco sheets were added at a ratio of 20%-25% and rolled into cigarettes, with each cigarette having a filling weight of approximately 0.8 g. The rolled cigarettes were placed in a constant temperature and humidity chamber at (22±1)°C and 60%±2% relative humidity for 48 h to equilibrate. Evaluation was conducted according to relevant enterprise standards. The results showed that the tobacco sheets prepared from the stem extract fermented by the aroma-producing Candida heterotrophic strain improved the aroma quality and quantity of cigarettes, increased sweetness, reduced off-flavors and irritation, enhanced aftertaste, and significantly improved sensory quality.

[0092] Example 8: Application of aroma-producing Candida TC5 in electronic cigarettes

[0093] The propylene glycol extract from Example 3 was added to the e-cigarette base solvent (40% glycerin + 60% propylene glycol) at a ratio of 10-20% (mass fraction) to obtain e-cigarette liquid. 0.5 g of e-cigarette liquid was added to each cartridge, and sensory quality evaluation of the e-cigarette samples was conducted. The results showed that the prepared e-cigarette had a fragrance similar to acacia and jasmine, and the aerosol produced had reduced off-flavors, enhanced sweetness, a clean aftertaste, and good vaping quality.

[0094] Example 9: Heterogeneous Candida TC5 enhances the aroma quality of cigarette tobacco leaves

[0095] Following step 1 in Example 2, the aroma-producing Candida albicans fermentation broth was obtained in advance. After centrifugation, the cells were redissolved and mixed with sterile water, resulting in a bacterial concentration of 10. 6 -10 8 CFU / mL, based on the tobacco leaf moisture content, was sprayed onto the tobacco leaves during the re-drying process at a ratio of 5%-10% by weight. After replenishing moisture and vacuum rehydration to a moisture content of 20%, the leaves were fermented at 30℃ for 24-36 hours, followed by re-drying at 100-120℃ to reduce the moisture content to 12%. The re-dried tobacco leaves were then shredded and rolled into sample cigarettes for smoking evaluation. The results showed that the use of aroma-producing heterologous Candida strains in the re-drying of cigarette tobacco leaves significantly improved the aroma quality and quantity of cigarettes, reduced irritation, increased sweetness, and enhanced comfort.

[0096] Example 10 Application of heterologous Candida TC5 strain in enhanced cigar fermentation

[0097] (1) Preparation of enhanced fermentation agent

[0098] The purified aroma-producing heterologous Candida was inoculated into a PDA liquid culture medium to obtain an aroma-producing heterologous Candida fermentation liquor, and after centrifugation, the bacterial cells were redissolved and mixed uniformly with sterile water, and the concentration of the bacterial liquor was 10 6 -10 8 CFU / mL.

[0099] (2) Strengthening the addition of a fermentation agent

[0100] The specific operation is to fill a spray bottle with 20% of the mass of cigar tobacco with a strengthening fermentation agent, spread the cigar tobacco, and uniformly spray the prepared strengthening fermentation agent onto the surface of the tobacco. After uniform spraying, the cigar tobacco is placed in a self-sealing bag for moisture recovery.

[0101] (3) Preparation of a fermentation agent for a cigar strengthening fermentation process

[0102] The tobacco in (2) is wrapped with cotton cloth and subjected to stacking and temperature variation fermentation in a constant-temperature and constant-humidity fermentation box. The process specifically includes four stages: initial 0-5 days, 30°C constant temperature; 5th to 15th day, 35°C constant temperature; 15th to 25th day, 40°C constant temperature; and last 25th to 30th day, 45°C constant temperature. The relative humidity range is maintained at 60%-80% in each stage. The control is without the addition of a fermentation agent.

[0103] (4) Sensory evaluation of cigar tobacco

[0104] The evaluation results show that the sensory quality of the cigar tobacco samples obtained by the fermentation processes of (2) and (3) is increased compared with the control (Table 4). In terms of aroma characteristics, the aroma amount, richness, and maturity are improved. In terms of smoke characteristics and aftertaste characteristics, they are also increased compared with the control. After fermentation, the honey sweet aroma and floral aroma of the cigar are prominent compared with the control, and the cigar style is highlighted.

[0105] Table 4 Sensory quality evaluation of cigar tobacco with the addition of aroma-producing heterologous Candida TC5 strengthening fermentation agent

[0106]

[0107] Although the present application has been disclosed with reference to the preferred embodiments above, it is not intended to limit the present application, and any person skilled in the art can make various modifications and modifications without departing from the spirit and scope of the present application. Therefore, the protection scope of the present application should be defined by the claims.

Claims

1. A strain of Candida zeylanoides TC5, characterized in that, The heterologous Candida sp. TC5, classified as Candida sp. (Candida Candida allociferrii ), has a preservation number of CCTCC NO: M 20241402, and was preserved in the China Center for Type Culture Collection on July 1, 2024, at the Wuhan University in Wuhan, China. The aroma-producing substance of the aroma-producing heterologous Candida sp. TC5 includes ester substances: methyl benzoate and methyl anthranilate.

2. A composition characterized in that, The active ingredient of the composition comprises the bioinoculant of the heterologous Candida TC5 of claim 1 or the fermentation broth of the heterologous Candida TC5.

3. The composition of claim 2, wherein, The bioinoculant of the heterologous Candida TC5 comprises live cells of the heterologous Candida TC5, freeze-dried bodies of the heterologous Candida TC5 obtained by freeze-drying, or immobilized cells of the heterologous Candida TC5.

4. The composition of claim 3, wherein, The bioinoculant of the heterologous Candida TC5 further comprises bacillus, yeast and / or white rot fungus.

5. Use of the aroma-producing heterologous Candida TC5 of claim 1 or the composition of any one of claims 2-4 in the preparation of tobacco products.

6. Use according to claim 5, characterized in that, The tobacco products comprise tobacco flavor, reconstituted tobacco sheet, electronic cigarette liquid or cigarette tobacco.

7. The aroma-producing heterologous Candida TC5 of claim 1 in the directed production of a plurality of odor substances, including floral, mushroom, cocoa, and almond.

8. Use of the aroma-producing heterologous Candida TC5 of claim 1 or the composition of any one of claims 2-4 in the improvement of flavor quality of tobacco products.

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