Aroma-producing heterologous candida and application of aroma-producing heterologous candida in tobacco products

Abstract

The invention belongs to the technical field of agricultural microorganisms, and discloses aroma-producing heterologous candida and application thereof in tobacco products. The strain can meet the requirements of different tobacco products, the stress resistance of the strain is excellent, a strain fermentation solution can generate 54 characteristic aroma substances with higher content, a propylene glycol extracting solution can generate 52 characteristic aroma substances, and after flue-cured tobacco leaves are fermented, compared with untreated tobacco leaves, the aroma quality and the aroma quantity are increased, the irritation is reduced, and the sweet feeling is increased; the cigar tobacco leaves are fermented for 30 days, the comfort is improved, the aired cigar tobacco leaves can reach the smoking standard after being intensively fermented for 30 days, the agricultural fermentation time is shortened, the fragrance such as honey sweet fragrance, fresh sweet fragrance and flower fragrance of the fermented cigar tobacco leaves is improved, and the cigar style is manifested. The heterologous candida provided by the invention is an aroma-producing strain, can directionally improve the flavor quality of tobacco products, and has a very good application prospect in aroma enhancement and quality improvement of products such as cigarettes and cigars.

Description

Technical Field

[0001] The invention belongs to the technical field of agricultural microorganisms, and particularly relates to an aroma-producing heterologous Candida and application of the same in tobacco products. Background Art

[0002] As a global cash crop, tobacco products are evolving towards reducing harm, enriching flavor, and enhancing sensory comfort. Tobacco is the primary raw material for cigarettes. With its unique aroma and convenient consumption, cigarettes have become a core product for the traditional tobacco industry, meeting market demand. However, growing consumer demand for high-quality cigarettes has placed higher demands on technologies to enhance cigarette quality and flavor. Research has shown that the formation of characteristic cigarette flavor is closely linked to the metabolic regulation of the microbial community during the fermentation of tobacco raw materials, playing an irreplaceable role in the transformation of aroma components.

[0003] Microbial fermentation has great potential for enhancing the flavor and quality of cigarettes. This method involves directly inoculating aroma-producing microorganisms onto the surface of tobacco leaves to ferment and produce aroma, or using these microorganisms to prepare tobacco flavorings and then adding them to the tobacco for flavor enhancement. This method is not restricted by climate, has a short production cycle, and produces high-quality, diverse flavor substances. Currently, the main microorganisms used in tobacco and cigarette products are Clostridium ( Clostridium ), Micrococcus ( Micrococci ), Pseudomonas ( Pseudomonas putida ), Bacillus ( Bacillus ) and Saccharomyces cerevisiae ( Saccharomyces cerevisiae ). However, there's been less research on the application of Candida in improving tobacco flavor quality, with more applications in fermented foods (such as soy sauce, fermented bean paste, and fermented black beans). For example, prior art reports have reported using Candida SY152 in fermenting fermented bean paste or soy sauce, enhancing its mellow aroma and significantly improving its taste and flavor. Candida eichei has also been reported to enhance the fermentation of low-salt raw bean paste, improving its color, flavor, and typicality. And Candida tropicalis JSY83 has been used in the production of Daqu and Luzhou-flavor liquors, increasing both the yield and quality of high-quality products.

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

[0005] To solve the above problems, in view of the fact that there is no report on the preparation of flavors and fragrances based on aroma-producing heterologous Candida in the production of tobacco products such as cigarettes to improve the flavor quality, the present invention provides a heterologous Candida with aroma-producing function ( White allocyanine ) strains and their applications in tobacco products.

[0006] In the first aspect, the present invention provides a fragrance-producing heterologous Candida, which is classified as heterologous Candida Candida allociferrii TC5, with the deposit number of CCTCC NO: M 20241402, was deposited in the China Center for Type Culture Collection on July 1, 2024, and the deposit address is Wuhan University, Wuhan, China.

[0007] In the present invention, the monoclonal colonies of the heterologous Candida yeast are characterized by white, round, raised colonies with a relatively smooth and moist surface. They also exhibit fermentation characteristics such as high temperature resistance, high acidity, high nicotine tolerance, and directional aroma production. The heterologous Candida yeast has a wide temperature tolerance range and maintains good growth within the range of 15°C to 45°C, meeting the requirements of the entire fermentation production stage of cigar tobacco leaves.

[0008] In a second aspect, the present invention further provides a composition for improving the flavor quality of tobacco products, wherein the main active ingredients of the composition include the heterologous Candida biological agent, the fermentation broth of the heterologous Candida, or the propylene glycol extract of the heterologous Candida.

[0009] In one embodiment of the present invention, the heterologous Candida biological agent further comprises Bacillus, yeast and / or white rot fungi.

[0010] In a third aspect, the present invention further provides use of the aroma-producing heterologous Candida described in the first aspect of the present invention or the composition described in the second aspect of the present invention in the preparation of tobacco products.

[0011] In one embodiment of the present invention, the tobacco product is tobacco flavoring, tobacco extract, tobacco flakes, flue-cured tobacco leaves, electronic cigarette liquid or cigar tobacco leaves, etc.

[0012] In one embodiment of the present invention, the tobacco product is a tobacco flavoring, and the tobacco flavoring is prepared using the fermentation broth of the heterologous Candida or the propylene glycol extract of the heterologous Candida as a raw material.

[0013] In one embodiment of the present invention, the tobacco product is tobacco extract, and the tobacco extract is obtained by fermenting tobacco extract using the heterologous Candida.

[0014] In one embodiment of the present invention, the tobacco product is a reconstituted tobacco leaf sheet, which is prepared by fermenting the stem paste of the heterologous Candida using a papermaking method.

[0015] In one embodiment of the present invention, the tobacco product is an electronic cigarette liquid, and the electronic cigarette liquid is prepared using the propylene glycol extract of the heterologous Candida strain.

[0016] In one embodiment of the present invention, the tobacco product is a cigarette leaf, which is prepared by directly acting the heterologous Candida liquid on the cigarette leaf or spraying it on the surface of the tobacco leaf to be aged as a cigarette leaf aging agent to accelerate the aging of the cigarette, or applying it to the cigarette leaf during the leaf threshing and redrying stage, or spraying it on the tobacco leaf during the cigarette shredding stage.

[0017] In one embodiment of the present invention, the tobacco product is cigar tobacco leaves, which are subjected to enhanced fermentation by directly acting on the heterologous Candida liquid or spraying it as an enhanced fermentation agent on the surface of the cigar tobacco leaves to be fermented or adding it to the surface of the cigar tobacco leaves to be fermented by pressurized atomization.

[0018] In a fourth aspect, the present invention further provides the use of the aroma-producing heterologous Candida described in the first aspect of the present invention or the composition described in the second aspect in the directed production of various odor substances having floral, mushroom, cocoa, rose, almond, etc.

[0019] In one embodiment of the present invention, the odor substances include the following substances, Esters: methyl benzoate, methyl anthranilate, Alcohols: 1-octen-3-ol, 3-methyl-1-butanol and 2-ethylhexanol, etc. Aldehydes: Benzoin aldehyde, nonanal, Ketones: isophorone, acetophenone, geranyl acetone, Phenols: 2-methoxy-4-vinylphenol.

[0020] Compared with the prior art, the present invention has the following technical effects: (1) The aroma-producing heterologous Candida TC5 provided by the present invention can produce 54 characteristic aroma substances in the culture medium fermentation broth. The aroma substances with high production in a targeted manner include esters (methyl benzoate, methyl anthranilate, etc.) and ketones (acetophenone, geranyl acetone) with floral and fruity aromas, as well as alcohols (1-octen-3-ol, 3-methyl-1-butanol and 2-ethylhexanol, etc.) with mushroom, cocoa and rose flavors.

[0021] (2) When the aroma-producing heterologous Candida TC5 provided by the present invention is used for enhanced fermentation of flue-cured tobacco leaves, compared with untreated tobacco leaves, the aroma quality and aroma quantity are increased, the irritation is reduced, the sweetness is increased, and the comfort is improved.

[0022] (3) When the aroma-producing heterologous Candida TC5 provided by the present invention is used for enhanced fermentation production of cigars, compared with normal fermentation, the fermentation time of 30 days can meet the smoking standard and shorten the aging time. After the fermentation, the honey sweet aroma, fresh sweet aroma, floral aroma and other aromas of the cigar tobacco leaves are enhanced, the cigar style is highlighted, the richness and aftertaste are improved, and the sensory quality of the smoking is significantly higher than that of the non-enhanced fermented tobacco leaves. The main chemical components meet the national standard requirements for cigars. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The aroma-producing heterologous Candida of the present invention ( Candida allociferrii ) Colony morphology and microscopic morphology of TC5; A is the plate colony image, and B is the microscopic observation result of the strain.

[0024] Figure 2 The aroma-producing heterologous Candida of the present invention ( Candida allociferrii ) Phylogenetic tree of TC5 strain.

[0025] Figure 3 The aroma-producing heterologous Candida of the present invention ( Candida allociferrii ) Radar chart of sensory evaluation of the fermentation broth and propylene glycol extract of TC5 strain in cigarettes; A is the chart showing the functional improvement effect on cigarettes, and B is the chart showing the aroma improvement effect on cigarettes. DETAILED DESCRIPTION

[0026] The following describes the specific embodiments of the present invention with reference to the accompanying drawings. The experimental methods used in the examples are conventional methods unless otherwise specified; the materials and reagents used are commercially available unless otherwise specified.

[0027] The main chemical components of flue-cured tobacco and cigar leaves (total sugars, reducing sugars, total nitrogen, total alkaloids, potassium, and chloride) are tested using a continuous flow analyzer. Aroma-producing Candida fermentation broth and volatile flavor compounds in tobacco leaves are analyzed using gas chromatography-mass spectrometry (GC-MS). Sensory evaluation of cigarettes and cigars follows tobacco industry standards and relevant industrial enterprise standards.

[0028] 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 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.

[0029] PDA medium: potato extract 12.0 g / L, glucose 20.0 g / L, agar 14.0 g / L, dissolved in distilled water, divided into portions, and autoclaved at 121°C for 15 min before use.

[0030] Saccharification liquid culture medium: Mix raw rice and water (1:4 mass ratio), add liquefaction enzyme at a mass fraction of 2‰ of the raw rice and 1‰ of saccharification enzyme, saccharify and liquefy in a 60℃ water bath for about 4-6 hours, adjust the sugar content to 13 Brix, sterilize at 115℃ for 20 minutes, and cool for use.

[0031] Preparation of tobacco leaf culture medium: 3.0 g of cigar tobacco leaves were chopped, added with 50 mL of distilled water, shaken on a shaker at 200 rpm for 30-60 min, and sterilized by high pressure at 121°C for 20 min to obtain tobacco leaf culture medium.

[0032] Example 1 Screening and identification of strains (1) Sample pretreatment Weigh 5 g of fermented cigar tobacco leaves or aged flue-cured tobacco leaves, chop them into pieces, add them to 50 mL of 0.85% sterile saline, and place them in a stacked full-temperature shaking incubator with the temperature set at 30°C and the rotation speed at 150 r / min for 2 h.

[0033] (2) Dilute and coat the plate Take the sample supernatant in step 1 and dilute it with sterile water (10 -1 ~10 -8 ), take different concentration gradients of dilutions and spread them on solid culture medium so that the microorganisms can grow into single colonies on the culture medium, culture them on PDA solid culture medium, and then place them in a constant temperature and humidity incubator for 48 hours.

[0034] (3) Separation of three zones Based on the differences in morphological characteristics, different monoclonal colonies from step (2) were selected and three-zoned on a new PDA plate. The plate with the zones was inverted and incubated in a constant temperature and humidity incubator at 30°C for 48 h. After multiple separation and purification, multiple strains with aroma-producing effects were obtained. The aroma-producing strains that were initially screened were then inoculated into PDA liquid culture medium for enrichment culture for 48 h. The aroma-producing strains with the best aroma-producing effects were selected by smelling.

[0035] (4) Purification and preservation Repeat step (3) for 2-3 purifications until only monoclonal colonies with a single morphological characteristic are present on the plate. Then, inoculate the monoclonal colonies into liquid PDA culture medium and culture them in a shaker at 30°C and 150 r / min for 48 h. Then, mix the bacterial solution with glycerol at a ratio of 1:1 and place them in 2 mL bacterial preservation tubes. After numbering, store them at -80°C.

[0036] (5) Strain identification Select the strain preserved in step (4), streak it on PDA solid culture medium, place it in a constant temperature and humidity incubator at 30℃ for 48 hours, observe its colony morphology and observe its cell morphology under a biological microscope after staining with crystal violet, as shown in Figure 2. Figure 1 As shown, the morphological characteristics of the monoclonal colony are white, round, raised, smooth and moist, and have long silky pseudohyphae under the microscope. Monoclonal colonies on the plate were placed in sterile water, boiled at high temperature to extract DNA, amplified and sent for sequencing. The nucleotide sequence was obtained by ITS rDNA sequencing, as shown in SEQ ID NO.1, and compared with the NCBI database for analysis. Figure 2 As shown, the strain has 100% homology with heterologous Candida and can be identified as heterologous Candida. The strain was deposited in the China Center for Type Culture Collection on July 1, 2024, with the deposit number CCTCC NO: M 20241402, and the deposit address is Wuhan University, Wuhan, China.

[0037] (6) Strain tolerance analysis The pH, temperature and nicotine tolerance of strain TC5 were analyzed, and the results showed that the strain could grow at 15℃-45℃, pH 4-11 and nicotine content within 8000 mg / L. Example 2 Determination of Volatile Flavor Compounds in Fermentation Broth of Heterologous Candida Strain TC5 (1) Preparation of TC5 fermentation broth The heterologous Candida preserved in glycerol tubes was streaked to obtain a single clone. The purified TC5 was inoculated at a 2% (volume fraction) into 50 mL of saccharification liquid for fermentation. The supernatant was collected by centrifugation and poured into a brown glass bottle to obtain a Candida fermentation broth with a floral and sweet aroma.

[0038] (2) Determination of volatile flavor substances The volatile flavor compounds in the fermentation broth of heterologous Candida strains 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 mixture was extracted and tested.

[0039] (3) Analysis of volatile flavor compounds The volatile flavor compounds in the fermentation broth were qualitatively analyzed using the NIST14 database to determine the categories of volatile flavor compounds. Semi-quantitative analysis based on internal standards was performed to determine the relative contents of the main flavor compounds. The results are shown in Table 1.

[0040] Table 1 Volatile flavor compounds in TC5 fermentation broth Example 3 Determination of Volatile Flavor Compounds in the Propylene Glycol Extract of the Heterologous Candida Strain TC5 (1) Preparation of propylene glycol solvent extract The heterologous Candida preserved in the glycerol tube was streaked to obtain a single clone. The TC5 strain was inoculated and spread onto a PDA plate for culture. The colonies on the plate had a strong floral aroma. The strain was gently scraped off with an inoculating loop (2.0 g) and thoroughly mixed with 20 mL of propylene glycol. After standing for 12 h, the supernatant was centrifuged and poured into a brown glass bottle to obtain the heterologous Candida propylene glycol extract.

[0041] (2) Determination of volatile flavor substances 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).

[0042] (3) Analysis of volatile flavor compounds The volatile flavor compounds in the propylene glycol extract were qualitatively analyzed using the NIST14 database to determine the categories of volatile flavor compounds. Semi-quantitative analysis based on internal standards was performed to determine the relative contents of the main flavor compounds. The results are shown in Table 2.

[0043] Table 2 Volatile flavor substances in propylene glycol solvent extract Example 4 Determination of Volatile Flavor Compounds in Cigarette Dust Fermented by Heterologous Candida TC5 (1) Preparation of TC5 fermentation broth The heterologous Candida preserved in glycerol tubes was streaked to obtain single clones. The purified TC5 was inoculated into 50 mL of heterologous Candida smoke powder culture medium at a volume fraction of 2% and the mixture was stirred at 150 r·min. -1 , 30℃ shaking fermentation for 36 h, 4000 r·min -1 , centrifuge for 10 min, take the supernatant, pour it into a brown glass bottle, and obtain the heterologous Candida smoke powder culture medium.

[0044] (2) Determination of volatile flavor substances The volatile flavor substances in the culture medium of heterologous Candida smoked tobacco were determined by headspace solid phase microextraction-gas chromatography-mass spectrometry, as described in Example 2 (2).

[0045] (3) Analysis of volatile flavor compounds The volatile flavor compounds in the tobacco broth were qualitatively analyzed using the NIST14 database to determine the categories of volatile flavor compounds. Semi-quantitative analysis based on internal standards was performed to determine the relative contents of the main flavor compounds. The results are shown in Table 3.

[0046] Table 3 Volatile flavor substances in tobacco waste culture medium Example 5 Preparation of tobacco flavoring by heterologous Candida TC5 The purified TC5 was inoculated into 50 mL of saccharification liquid at a 2% inoculation rate, fermented for 36 h, and the supernatant was centrifuged as the fermentation liquid; the TC5 strain was inoculated and spread on a PDA plate, cultured at 30°C for 48 h, and then the strain (2.0 g) was gently scraped off with an inoculation loop and thoroughly mixed with 20 mL of propylene glycol. After standing for 12 h, the supernatant was centrifuged as the bacterial propylene glycol extract.

[0047] The sensory evaluation results of the cigarettes after injection of fermentation broth and propylene glycol extract are as follows Figure 3 As shown. Compared with the control, the sensory quality indicators of the injected cigarettes showed varying degrees of change. In terms of aroma characteristics, both effectively improved the aroma quality, volume, concentration, and permeability of the cigarettes, and reduced unpleasant odors. In terms of taste characteristics, both produced a refined and soft smoke. However, the fermentation broth slightly increased the irritation and residual properties of the smoke, and the propylene glycol extract also had a slight residual. This may be due to the presence of small amounts of bacterial metabolites such as protein in the extracts. Except for the two indicators of irritation and residual properties, the fermentation broth had a better effect on improving sensory properties than the propylene glycol extract. The fermentation broth exhibited a roasted and sweet aroma, while the propylene glycol extract exhibited a fresh, fruity, woody, floral, and sweet aroma, with the sweet aroma being the most prominent. The results show that the fermentation broth and propylene glycol extract can improve the aroma and taste characteristics of cigarettes and enhance the fragrance.

[0048] Example 6 Preparation of tobacco extract by heterologous Candida TC5 According to the method of step 1 in Example 2, the aroma-producing heterologous Candida fermentation broth was obtained in advance and centrifuged, and the cells were redissolved and mixed with sterile water. 200 g of commercially available tobacco extract was selected as the raw material and placed in a 500 mL triangular shake flask. After the heterologous Candida fermentation broth was inoculated at a ratio of 1%-10%, the mixture was cultured and fermented in a constant temperature and humidity incubator at 30°C for 3-5 days. After the fermentation, the tobacco extract obtained after fermentation was dissolved and diluted with sterile water, and then sprayed on unflavored cut tobacco at a dosage of 0.05-0.1%. The flavor quality was evaluated by professional judges according to the tobacco additive sensory evaluation method in accordance with the national standard GB 5606.4-2005 "Technical Requirements for Sensory Sensory of Cigarettes". The results showed that the aroma-producing heterologous Candida strain can significantly improve the quality of tobacco extract through fermentation, increasing the aroma, reducing the impurities, reducing the irritation, improving the comfort, and increasing the sweetness.

[0049] Example 7 Application of Aroma-Producing Heterologous Candida TC5 in Tobacco Sheets According to the method of step 1 in Example 2, the fermentation liquid of aroma-producing heterologous Candida was obtained in advance and centrifuged, and the bacteria were redissolved and mixed with sterile water. The concentration of the bacterial liquid was 10 6 -10 8 CFU / mL. The bacterial solution was inoculated into the stem paste at a dry matter mass fraction of 5%-10%, depending on the moisture content. The mixture was then fermented at 30°C for 1-12 h, coated onto a sheet substrate at a coating rate of 20%-50%, and dried at 90°C for 10 min. After rehydration to a moisture content of 10-13%, the tobacco flakes were shredded. Tobacco flakes were added at a ratio of 20%-25% to form cigarette sticks, with each stick containing approximately 0.8 g of filling. The rolled cigarette sticks were equilibrated in a constant temperature and humidity chamber at (22±1)°C and a relative humidity of 60%±2% for 48 h. Smoking tests were conducted according to relevant industry standards. The results showed that tobacco flakes prepared from stem paste fermented with aroma-producing Candida species improved the aroma quality and quantity of cigarettes, increased sweetness, reduced off-flavors and irritation, and enhanced aftertaste, significantly improving the sensory quality of the cigarettes.

[0050] Example 8 Application of aroma-producing Candida TC5 in electronic cigarettes The propylene glycol extract from Example 3 was added to an e-cigarette base solvent (40% glycerin + 60% propylene glycol) at a ratio of 10-20% by mass to produce e-cigarette liquid. 0.5 g of the liquid was added to each cartridge, and sensory quality evaluation of the e-cigarette samples revealed that the prepared e-cigarette exhibited aromas similar to those of Sophora japonica and jasmine. The aerosol produced had reduced aroma contaminants, a more sweet and mellow aroma, a clean aftertaste, and excellent puff quality.

[0051] Example 9: Heterologous Candida TC5 improves the aroma quality of cigarette tobacco leaves According to the method of step 1 in Example 2, the fermentation liquid of aroma-producing heterologous Candida was obtained in advance and centrifuged, and the bacteria were redissolved and mixed with sterile water. The concentration of the bacterial liquid was 10 6 -10 8 CFU / mL. Based on the tobacco leaf moisture content, the tobacco leaves were sprayed onto the tobacco leaves during the threshing and redrying process at a rate of 5%-10% by mass. Moisture was then added to the leaves after vacuum conditioning to a moisture content of 20%. The leaves were then fermented at 30°C for 24-36 hours and redryed at 100-120°C to reduce the moisture content to 12%. The redryed leaves were then cut and rolled into sample cigarettes for smoking evaluation. Results indicate that the use of aroma-producing Candida species in tobacco threshing and redrying significantly improves the aroma quality and quantity of cigarettes, reduces irritation, increases sweetness, and enhances comfort.

[0052] Example 10 Application of Heterologous Candida Strain TC5 to Enhance Cigar Fermentation (1) Preparation of enhanced starter culture The purified aroma-producing heterologous Candida was inoculated into PDA liquid culture medium to obtain aroma-producing heterologous Candida fermentation liquid. After centrifugation, the bacteria were redissolved and mixed with sterile water. The bacterial liquid concentration was 10 6 -10 8 CFU / mL.

[0053] (2) Addition of enhanced fermentation agent The specific operation is to put 20% of the fortified fermentation agent by weight of cigar tobacco leaves into a spray bottle, spread out the cigar tobacco leaves, and evenly spray the prepared fortified fermentation agent onto the surface of the tobacco leaves. After spraying evenly, put the cigar tobacco leaves into a ziplock bag to allow them to regain moisture.

[0054] (3) Preparation of starter cultures for cigar enhanced fermentation process The tobacco leaves from (2) were wrapped in cotton cloth and placed in a constant temperature and humidity fermentation chamber for stacked variable temperature fermentation. The process consisted of four stages: initial constant temperature of 30°C for days 0-5; constant temperature of 35°C for days 5-15; constant temperature of 40°C for days 15-25; and constant temperature of 45°C for days 25-30. Relative humidity was maintained in the range of 60%-80% during each stage. The control was a fermentation chamber without the addition of a starter agent.

[0055] (4) Sensory evaluation of cigar tobacco leaves The results of the smoking test showed that the sensory quality of the cigar tobacco samples obtained by fermentation using the fermentation processes (2) and (3) was improved compared with the control (Table 4). In terms of aroma characteristics, the aroma volume, richness and maturity were improved. The smoke characteristics and aftertaste characteristics were also increased compared with the control. After the fermentation, the honey sweetness and floral aroma of the cigar were more prominent than the control, and the cigar style was highlighted.

[0056] Table 4 Sensory quality evaluation of cigar tobacco leaves supplemented with aroma-producing Candida TC5 fortified starter Although the present invention has been disclosed above in terms of preferred embodiments, it is not intended to limit the present invention. Anyone familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the definition of the claims.

Claims

1. A fragrance-producing heterologous Candida species, characterized in that: The heterologous Candida is classified as heterologous Candida Candida allociferrii TC5, with the deposit number of CCTCC NO: M 20241402, was deposited in the China Center for Type Culture Collection on July 1, 2024, and the deposit address is Wuhan University, Wuhan, China.

2. A composition, characterized in that The main active ingredients of the composition include the biological agent of the heterologous Candida according to claim 1, the fermentation broth of the heterologous Candida or the propylene glycol extract of the heterologous Candida.

3. The composition according to claim 2, characterized in that The heterologous Candida biological agent includes living cells of the heterologous Candida, a lyophilized product obtained by freeze-drying the heterologous Candida, or immobilized cells of the heterologous Candida.

4. The composition according to claim 3, characterized in that The heterologous Candida biological agent also includes Bacillus, yeast and / or white rot fungi.

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

6. The use according to claim 5, characterized in that The tobacco products include tobacco flavorings, tobacco extracts, reconstituted tobacco leaf sheets, electronic cigarette liquids, cigarette tobacco leaves and / or cigar tobacco leaves.

7. Use of the aroma-producing heterologous Candida according to claim 1 in the directed production of multiple odor substances.

8. The use according to claim 7, characterized in that The odor substances include flower scent, mushroom scent, cocoa scent, rose scent, and almond scent.

9. Use of the aroma-producing heterologous Candida according to claim 1 or the composition according to any one of claims 2 to 4 in improving the flavor quality of tobacco products.

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