Aspergillus cephalosporium JDN-1 derived from mould on surface of pit top mud, method for inhibiting growth of aspergillus cephalosporium JDN-1 and application
By studying the biological characteristics of mold on the surface of the cellar mud and using temperature, anaerobic and pH adjustment methods to inhibit the growth of Aspergillus cefosfos, the adverse effects of mold on the fermentation of white wine were solved, and the cellar mud was realized Extend the use time of mud and reduce costs.
Patent Information
- Application Number
- CN202411315115.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-20
- Publication Date
- 2024-12-13
AI Technical Summary
The prior art is difficult to effectively inhibit the growth of mold on the surface of the cellar top mud, resulting in adverse effects during the fermentation process of the white wine, and the cost of replacing the cellar top mud or strengthening the management is high and the effect is limited.
By studying the biological characteristics of mold on the surface of the mud on the cellar, Aspergillus cefosfos was isolated and its growth was inhibited by regulating temperature, anaerobic, pH and lethal temperature, which specifically included adjusting temperature outside the range of 10-40℃ , inhibit aerobic bacteria growth under anaerobic conditions, adjust the pH to 3-4 to inhibit colony growth, and treat the water bath at 65°C for 10 minutes as the lethal temperature.
It effectively extends the use time of the cellar top mud, avoids the adverse effects of mold growth on the brewing of white wine, and provides a fast and economical method to inhibit the growth of mold on the surface of the cellar top mud.
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Figure CN119120214A_ABST
Abstract
Description
Technical Field
[0001] The present application relates to the technical field of research on mold growth on the surface of cellar roof mud, and in particular to a type of Aspergillus cephalosporin JDN-1 derived from mold on the surface of cellar roof mud, and a method and application of inhibiting its growth. Background Art
[0002] The cellar top mud, also known as the capping mud, is a layer of mud sealed on the surface after the mash enters the pool. It plays a role in isolating the cellar from the outside world during the cellar fermentation process, creating an independent anaerobic system for the fermentation of liquor. Due to the influence of gravity, material movement, etc., some microorganisms and nutrients contained in the cellar top mud will gradually penetrate into the mash during the cellar fermentation process and affect the fermentation of liquor. Normal cellar top mud has a small amount of bacteria or no bacteria growing on the surface. If it grows on a large area, it will produce a large amount of metabolites, which will change the material composition of the cellar top mud. As time goes by, these metabolites will penetrate into the cellar top mud and change the changes in the microorganisms and nutrients in the cellar top mud, thereby affecting the normal fermentation of liquor. Therefore, keeping the surface of the cellar top mud free of mold during the production process plays an important role in maintaining the normal brewing of liquor.
[0003] At present, when the cellar roof mud produces a situation where mold grows rampantly, there is no other effective way to prevent the growth of mold except replacing the cellar roof mud or strengthening management. Replacing the cellar roof mud increases costs, and strengthening management has little effect. Therefore, it is urgent to find a fast and economical adjustment method to inhibit the growth of mold. Summary of the invention
[0004] In view of the deficiencies in the prior art, the present application provides a cephalosporin Aspergillus JDN-1 derived from the mold on the surface of pit roof mud, a method for inhibiting its growth and an application thereof.
[0005] In order to solve the problem of mold growth on the surface of cellar roof mud, this application starts from the bacteria growing on the surface of cellar roof mud, explores the causes of its generation by studying its composition and its differences with growth conditions, raw materials, etc., and studies the growth characteristics of single strains; then, based on the causes of colony generation and its growth conditions, targeted changes are made to the production and maintenance of cellar roof mud, so as to achieve the purpose of controlling the growth of bacteria on the surface of cellar roof mud and improving the fermentation environment of liquor.
[0006] In a first aspect, the present application provides a cephalosporin fungus, using the following technical solution: A cephalosporin Aspergillus cejpii isolated from the surface of pit mud was named Aspergillus cejpii JDN-1. Aspergillus cejpii JDN-1 was deposited in the China Center for Type Culture Collection on August 16, 2024. The deposit address is: China Center for Type Culture Collection, Wuhan University, Wuchang District, Wuhan City, Hubei Province; the deposit number is CCTCC NO: M 20241807.
[0007] Aspergillus cejpii JDN-1 was isolated from the top mud of the sesame-flavored cellar and the top mud of the strong-flavored cellar. Its morphology on the Bengal red medium is as follows: the colonies are light red, radiating, with concentric circles, light yellow in the middle, with granular and hyphae on the surface; the spores are pink under the microscope. From the morphological point of view, the fungi are all Aspergillus colonies.
[0008] In a second aspect, the present application provides a method for inhibiting the growth of mold on the surface of cellar top mud, using the following technical scheme: a method for inhibiting the growth of mold on the surface of cellar top mud, wherein the mold on the surface of the cellar top mud is Aspergillus cephalosporin JDN-1; the inhibition method is to inhibit the growth of mold on the surface of the cellar top mud by using at least one of the following methods: (1) Adjust the temperature; (2) Anaerobic; (3) Adjust pH.
[0009] (4) Lethal temperature.
[0010] Preferably, the temperature in step (1) is adjusted to <10°C.
[0011] Preferably, in step (1), the temperature is adjusted to > 40°C.
[0012] From the biological characteristics of the fungus, we know that Aspergillus cephalosporin JDN-1 can grow at temperatures between 10 and 40°C, and the colonies will not grow below 10°C or above 40°C. Therefore, the growth and reproduction of Aspergillus cephalosporin JDN-1 can be inhibited under conditions below 10°C or above 40°C.
[0013] Preferably, the anaerobic condition in step (2) is a completely sealed state.
[0014] The biological characteristics of the fungus show that Aspergillus cephalosporin JDN-1 is an aerobic bacterium and its growth and reproduction can be inhibited under anaerobic conditions.
[0015] Preferably, in step (3), the pH is adjusted to 3-4.
[0016] According to the biological characteristics of the fungus, Aspergillus cephalosporin JDN-1 can grow in the pH range of 4-10, and the colonies do not grow at a pH of 3. Therefore, adjusting the pH to 3-4 can inhibit the growth and reproduction of Aspergillus cephalosporin JDN-1.
[0017] Preferably, the lethal temperature in step (4) is 65°C.
[0018] The biological characteristics of the fungus show that the colonies of Aspergillus cephalosporin JDN-1 can grow when treated in a water bath at 50-60°C for 10 minutes, but stop growing after being treated in a water bath at 65°C for 10 minutes. Therefore, the lethal temperature of Aspergillus cephalosporin JDN-1 is determined to be 65°C for 10 minutes.
[0019] In a third aspect, the present application provides a method for inhibiting mold growth on the surface of cellar top mud in an industrial winemaking process, using the following technical solution: A method for inhibiting mold growth on the surface of cellar top mud is used in an industrial winemaking process, wherein the mold on the surface of the cellar top mud is Aspergillus cephalosporin JDN-1, and at least one of the following methods is used to inhibit the growth of mold on the surface of the cellar top mud: (1) Anaerobic; (2) The ratio of the thickness of the pit top mud to the height of the fermentation container should be controlled to be ≥10%; (3) Expose the loess in the cellar top mud to control the water content of the loess in the cellar top mud.
[0020] Preferably, the anaerobic condition in (1) is in a completely sealed state.
[0021] By adopting the above technical solution, no bacteria will grow on the cellar top mud when it is in a completely sealed state. As the number of holes on the surface of the plastic sheet increases, more bacteria will grow on the cellar top mud, and the growing parts will all grow around the holes, which confirms that the mold growing on the surface of the cellar top mud is aerobic bacteria and its growth can be inhibited by anaerobic conditions.
[0022] When the thickness of the cellar top mud accounts for 10% of the height of the fermentation container, there is no mold growth on the plastic cloth on the surface of the cellar top mud in the absence of holes. This indicates that the thickness of the cellar top mud should not be too small in production, and the ratio of its thickness to the height of the fermentation container should be ≥10%, which can effectively inhibit the growth of mold on the surface of the cellar top mud.
[0023] Preferably, in step (3), the pit top mud includes old mud and loess; The specific steps of sun-drying the cellar top mud are as follows: sun-drying the loess in the cellar top mud, and controlling the water content of the loess in the cellar top mud to ≤11%; By adopting the above technical solution, after the loess is exposed to the sun, the moisture content of the loess is ≤11%, which can inhibit the growth of mold on the surface of the pit top mud.
[0024] In summary, the present application includes at least one of the following beneficial technical effects: The present application discloses a cephalosporin Aspergillus JDN-1 derived from mold on the surface of cellar roof mud, a method for inhibiting its growth and its application. The method can avoid the adverse effects of the growth of mold on the surface of cellar roof mud on liquor brewing and effectively prolong the service life of the cellar roof mud. BRIEF DESCRIPTION OF THE DRAWINGS
[0025] Figure 1 The colony and microscopic morphology of Aspergillus cephalosporin JDN-1; Figure 2 This is the phylogenetic tree of Aspergillus cephalosporin JDN-1 strain constructed based on ITS sequences; Figure 3 The effect of temperature on the colony diameter of Aspergillus cephalosporin JDN-1; Figure 4 The effect of pH on the colony diameter of Aspergillus cephalosporin JDN-1; Figure 5 It is a comparison diagram of the test cellar and the control cellar in Example 4. DETAILED DESCRIPTION
[0026] The technical solution of the present application is further illustrated by specific embodiments below. The specific embodiments do not limit the protection scope of the present application; some non-essential modifications and adjustments made by others based on the concept of the present application still fall within the protection scope of the present application.
[0027] The raw materials involved in this application are all commercially available products, and the application is further described in detail below in conjunction with the embodiments and comparative examples.
[0028] Example 1: Isolation and identification of mold 1. Isolation and identification of mold The bacteria on the surface of the cellar top mud were isolated by a culturable method, and their molecular biological identification was carried out through morphological observation and ITS sequence analysis of single strains.
[0029] 1. Materials and reagents 1.1 Test Materials The test materials are the top mud from the place where the fungi grow in the cellars of Luzhou-flavor liquor and the top mud from the place where the fungi grow in the cellars of sesame-flavor liquor.
[0030] 1.2 Culture medium and reagents Agar: Beijing Aoboxing Biotechnology Co., Ltd.; Fungal genomic DNA rapid extraction kit, Taq PCR Mix: Sangon Biotech (Shanghai) Co., Ltd.; Other reagents were of domestic analytical grade.
[0031] 1.3 Preparation of culture medium PDA culture medium: Peel 200g of peeled potatoes and boil them in water. Filter the filtrate through 8 layers of gauze and adjust the volume to 1L. Add 20g of glucose and 20g of agar, sterilize at 121℃ for 20 minutes, and pour the culture medium into a flat dish after it cools down for later use.
[0032] Rose Bengal culture medium: Weigh 41.5 g of Rose Bengal culture medium, add 1 L of deionized water, sterilize at 121°C for 20 minutes, and pour into a plate after the culture medium cools down for later use.
[0033] 2. Experimental methods and results 2.1. Isolation and purification of mold Take 25g of mold-containing pit top mud, add 225ml of sterile water, and mix in a homogenizer. Perform 10-fold gradient dilution under the clean bench, use a pipette to absorb 0.1ml of the dilution and apply it to the PDA medium. After evenly applying, place the culture dish in a 30℃ incubator and culture until the colony grows. Pick a single strain and inoculate it in the PDA medium for purification.
[0034] 2.2 Morphological observation of mold Colony observation: The mold purified in step 2.1 was inoculated into PDA culture medium using the spot-grafting method in an ultra-clean workbench and placed in a 30°C incubator for 5 days, during which the colony morphology was observed.
[0035] Cell observation: Place a glass slide in a culture dish, drop PDA culture medium on the glass slide, wait for it to solidify, and inoculate the mold purified in step 2.1 at the edge of the culture medium. After culturing in a 30°C incubator for 5 days, place the glass slide under a microscope to observe the morphology of the bacteria and spores.
[0036] The results showed that a fungus was isolated from the top mud of the sesame-flavored cellar and the top mud of the strong-flavored cellar, named as Aspergillus cephalosporin JDN-1; its morphology on Bengal red medium was as follows: Figure 1 As shown in the figure, the colonies are light red, radiate, have concentric circles, are light yellow in the middle, have granules on the surface, and have hyphae; the spores are pink under the microscope. From the morphological point of view, the fungi are all Aspergillus colonies.
[0037] 2.3. Molecular biological identification of strains The fungal genomic DNA extraction kit was used to extract the genomic DNA of the strain. The universal primers were used for PCR amplification. The universal primer sequences are as follows: The sequence of ITS1 is: 5'-TCCGTAGGTGAACCTGCGG-3'; as shown in SEQ ID No.2; the sequence of ITS4 is: 5'-TCCTCCGCTTATTGATATGC-3'; as shown in SEQ ID No.3.
[0038] The amplification procedure is: ~Pre-denaturation: 98℃, 5min; ~ Denaturation: 98℃, 30s; ~ Annealing: 55°C, 30s; ~ Extension: 72℃, 30s; ~The above three steps (denaturation, annealing, extension) need to be cycled 30 times; ~ Post-extension: 72°C, 10 min; ~Storage: 4°C, 10 min; The PCR products were detected by agarose gel electrophoresis and sent to Sangon Biotech (Shanghai) Co., Ltd. for sequencing. The obtained sequences were searched for homologous sequences using the BLAST program in NCB, and sequences with similarity greater than 99% and closely related genera were downloaded. Multiple sequence alignment was performed using MEGA-X software, and a phylogenetic tree was constructed using the neighbor-joining method.
[0039] The results showed that the gene sequence of the bacterium was shown in SEQ ID No. 1, and the evolutionary tree constructed after sequencing was as follows: Figure 2 As shown, JDN-1 and Aspergillus cejpii have the highest similarity, both reaching more than 100%, indicating that these fungi are Aspergillus cejpii, which belongs to the phylum Ascomycota, subclass Eurotiomycetes, family Aspergillus, and genus Aspergillus.
[0040] Example 2: Analysis of biological characteristics of mold Study the growth characteristics of each strain, such as tolerance to acid, alkali, temperature, etc., and find out the conditions that can inhibit the growth of the strain.
[0041] 1. Test strains The fungus Aspergillus cephalosporin JDN-1 was isolated from the surface of the pit top mud.
[0042] 2. Test methods 2.1. Strain activation Aspergillus cephalosporin JDN-1 was inoculated into PDA medium and cultured at 30°C for 5 days.
[0043] 2.2 Effect of temperature on strain growth Use a sterile cork puncher to punch out several activated mold cakes with a diameter of 6 mm at the edge of the colony (the activated strain in 2.1 in this example) and inoculate them on PDA culture medium. Place them in incubators at 5°C and 10°C, 15°C, 20°C, 25°C, 30°C, 35°C, 40°C, and 45°C, respectively, and culture them. On the 5th day, the colony diameter was measured by the cross method and the growth status of the strain was recorded. Each treatment was repeated 3 times.
[0044] The results show that: Figure 3It can be seen that Aspergillus cephalosporin JDN-1 can grow at temperatures between 10 and 40°C. The colonies will not grow below 10°C or above 40°C. Its optimal growth temperature is 30°C. After 5 days of culture at the optimal temperature, the colony diameter reaches 86 mm, which is significantly higher than other culture temperatures.
[0045] The biological characteristics of the fungus show that the growth and reproduction of Aspergillus cephalosporin JDN-1 can be inhibited at temperatures below 10°C or above 40°C.
[0046] 2.3 Effect of pH on the growth of strains The pH of the PDA medium was adjusted to 8 gradients of 3, 4, 5, 6, 7, 8, 9, and 10 with lactic acid and NaOH, respectively. The 6 mm activated mold cake (the activated strain in 2.1 in this example) was inoculated on the PDA medium with different pH values and cultured in an incubator at 28°C. The colony diameter was measured on the 5th day and the growth status of the strain was recorded.
[0047] The results show that: Figure 4 It can be seen that strain JDN-1 can grow in the pH range of 4-10, with the optimal growth pH of 7 and the colony diameter at the optimal pH of 82 mm. It is worth noting that the colonies do not grow at a pH of 3, indicating that the growth of these bacteria is not conducive to an acidic environment. It can be seen that lowering the pH of the cellar top mud can help inhibit the growth of mold without affecting the normal fermentation of the cellar.
[0048] According to the biological characteristics of the mold, when the pH is adjusted to 3-4, the growth and reproduction of Aspergillus cephalosporin JDN-1 can be inhibited.
[0049] 2.4 Effect of anaerobic culture on the growth of strains The 6 mm activated mold cake (the activated strain in 2.1 in this embodiment) was inoculated onto a PDA culture dish, one part of the culture dish was sealed with a plastic tape and placed in an anaerobic bag, and the other part of the culture dish was covered with a lid and placed in a 28° C. incubator for culture. The culture dishes were checked for bacterial growth on the 5th day.
[0050] The results showed that: as shown in Table 1, when the culture dish was completely sealed, Aspergillus cephalosporin JDN-1 did not grow, and the strain in the unsealed culture medium grew, indicating that Aspergillus cephalosporin JDN-1 was an aerobic bacterium and complete sealing could inhibit the growth of Aspergillus cephalosporin JDN-1.
[0051] The biological characteristics of the fungus show that Aspergillus cephalosporin JDN-1 is an aerobic bacterium. Under anaerobic conditions, the growth and reproduction of Aspergillus cephalosporin JDN-1 can be inhibited.
[0052] Table 1 Effects of anaerobic culture on strains Note: + represents colony growth, - represents colony no growth 2.5 Effect of lethal temperature on the growth of strains The activated mold cake with a diameter of 6 mm (the activated strain in 2.1 in this embodiment) was placed in a sterile centrifuge tube and placed in a constant temperature water bath at 50°C, 55°C, 60°C, 65°C, 70°C, and 75°C for 10 min, respectively, and then quickly cooled. The cake was taken out and inoculated on a PDA plate and cultured in an incubator at 28°C. The colony diameter was measured on the 5th day and the growth status of the strain was recorded.
[0053] The results showed that the colonies of Aspergillus cephalosporin JDN-1 could grow after being treated in a water bath at 50-60℃ for 10 minutes, but stopped growing after being treated in a water bath at 65℃ for 10 minutes. Therefore, the lethal temperature of Aspergillus cephalosporin JDN-1 was determined to be 65℃.
[0054] Example 3: Simulation test of fungus growth in pit top mud According to the origin, growth characteristics, etc. of the strain, targeted measures can be selected in combination with production to hinder its growth and reproduction.
[0055] 1. Test materials The cellar roof mud, loess and mash are all taken as materials used in the production workshop.
[0056] 2. Test methods 2.1 Effects of different sealing conditions on bacterial growth in pit top mud According to the cellar ratio, 1 kg of mash was placed in a foam box, a 4 cm layer of cellar top mud was added, and then covered with a layer of plastic cloth to completely seal it. After 1, 3, and 5 holes (size 0.5 cm × 0.5 cm) were poked through the box respectively, the box was placed in a 25°C environment and cultured for 30 days. During this period, the growth of bacteria on the surface of the cellar top mud was observed.
[0057] The results show that: as shown in Table 2, no bacteria grow on the cellar top mud when it is in a completely sealed state. As the number of holes on the plastic sheet increases, more bacteria grow on the cellar top mud, and the growing areas all grow around the holes, which confirms that the mold growing on the surface of the cellar top mud is aerobic bacteria, and anaerobic conditions can inhibit its growth.
[0058] Table 2 Effects of different sealing conditions on the growth of bacteria in the pit top mud serial number Hole situation Sealed state Bacteria growth A1 0 Hole seal - A2 1 hole Not sealed + A3 3 holes Not sealed ++ A4 5 holes Not sealed +++ Note: + represents bacteria growth, - represents no bacteria growth 2.2 Effect of different pH values on bacterial growth in pit top mud Taking the normally used cellar top mud as the control, lactic acid and NaOH were used to adjust the pH of the cellar top mud to 3.5, 4.5, 4, 5, 6 and 7 respectively; according to the cellar ratio, 1 kg of mash was placed in a foam box, and a 4 cm thick layer of cellar top mud was added, and then covered with a layer of plastic cloth, and placed in a 25°C environment for incubation for 30 days, during which the growth of bacteria on the surface of the cellar top mud was observed.
[0059] The results show that: as shown in Table 3, in the unsealed state, when the pH is 4, no bacteria grow on the surface of the cellar top mud, and when the pH is greater than 4.5, mold grows on the surface of the cellar top mud, indicating that in the presence of a small amount of oxygen, when the cellar top mud pH is ≤ 4, mold growth can be inhibited. The pH of the cellar top mud currently used in production is between 6 and 7, and the pH deviation of the cellar top mud with a pH of 4 is large from the pH of the cellar top mud currently used in production. Under the influence of gravity and the free diffusion of molecules, if the pH of the cellar top mud is adjusted too low rashly, the acidic group molecules will penetrate into the cellar, changing the pH of the cellar, affecting the growth and metabolism of liquor brewing microorganisms, and further affecting the liquor production and aroma production functions. Therefore, the method of inhibiting mold growth by adjusting the pH of the cellar top mud is not suitable for industrial winemaking.
[0060] Table 3 Effect of different pH on the growth of bacteria in the pit top mud serial number Hole situation pH Bacteria growth B1 3 holes 3.5 - B2 3 holes 4 - B3 3 holes 4.5 + B4 3 holes 5 ++ B5 3 holes 6 ++ B6 3 holes 7 +++ Note: + represents bacteria growth, - represents no bacteria growth 2.3. Effect of cellar roof mud thickness on fungus growth Taking normally used cellar roof mud as the control, according to the cellar proportion, 1 kg of mash was placed in a foam box, and a layer of 2 cm, 4 cm, 6 cm, 8 cm, and 10 cm thick cellar roof mud was added respectively, and then covered with a layer of plastic cloth, and placed in a 25°C environment for incubation for 30 days. During this period, the fungus growth on the surface of the cellar roof mud was observed.
[0061] The results show that: as shown in Table 4, in the case of poor sealing (i.e., there are holes on the surface of the plastic sheet), regardless of the thickness of the cellar top mud, the surface of the cellar top mud will grow mold, which is related to the fact that the mixing of oxygen promotes the growth of mold. When the thickness of the cellar top mud and the height of the fermentation container are ≤9%, the plastic sheet on the surface of the cellar top mud also has mold growth without holes, indicating that when the cellar top mud is in a sealed state, a small amount of mold will grow if the thickness of the cellar top mud is too small. This is because the mold in the fermented grains can pass through the cellar top mud and cause surface growth when the thickness of the cellar top mud is too small. When the thickness of the cellar top mud and the height of the fermentation container are 10%, there is no mold growth on the plastic sheet on the surface of the cellar top mud without holes. It can be seen that the thickness of the cellar top mud should not be too small in production, and its thickness should be ≥10% of the height of the fermentation container, which can effectively inhibit the growth of mold on the surface of the cellar top mud.
[0062] Table 4 Effect of different thicknesses on the growth of bacteria in the pit top mud Note: + represents bacteria growth, - represents no bacteria growth 2.4 Effect of raw materials on the growth of bacteria in pit top mud The raw materials of the cellar top mud are old mud (i.e. the cellar top mud that has been fermented in the previous round) and loess. The old mud and loess are exposed to the sun for one day. The water content of the old mud is reduced from 33% to 25% after exposure, and the water content of the loess is reduced from 15% to 11%. After the loess and old mud are mixed in a ratio of 1:2, an appropriate amount of water is added (so that the water content of the cellar top mud reaches 35±3%) to make the cellar top mud. According to the cellar ratio, 1kg of fermented grains is placed in a foam box, a layer of 4cm of cellar top mud is added, and then a layer of plastic cloth is covered, and it is placed in a 25℃ environment for cultivation. During this period, the growth of bacteria on the surface of the cellar top mud is observed.
[0063] It can be seen from Table 5 that loess alone does not grow bacteria, and loess does not grow bacteria after being exposed to the sun and mixed with old mud. Old mud grows bacteria after being exposed to the sun and mixed with unexposed loess. This shows that old mud can provide nutrients to the mold in the loess. Eliminating the mold in the loess is of great significance to preventing bacteria from growing in the cellar top mud.
[0064] Table 5 Effect of raw materials on the growth of bacteria in pit top mud serial number Old mud state Loess state Bacteria growth D1 Unsunned / + D2 / Unsunned - D3 Unsunned Unsunned +++ D4 Sun Sun - D5 Unsunned Sun - D6 Sun Unsunned +++ Note: + represents bacteria growth, - represents no bacteria growth In order to verify the effect of the water content of loess in the cellar top mud on the growth of the cellar top mud, the old mud and loess were exposed to the sun for one day, and the water content of the loess after exposure was controlled to be 15%, 13%, 12%, 11%, 10%, 9%, and 8%, respectively. The loess with different water contents were mixed with the old mud at a ratio of 1:2, and then an appropriate amount of water was added (to make the water content of the cellar top mud reach 36%) to make the cellar top mud. According to the cellar ratio, 1kg of fermented grains was placed in a foam box, a layer of 4cm of cellar top mud was added, and then a layer of plastic cloth was covered, and it was placed in a 25℃ environment for cultivation, and the growth of bacteria on the surface of the cellar top mud was observed during the period.
[0065] It can be seen from Table 6 that the water content of loess has a certain influence on the growth of mold in the cellar top mud. When the water content of loess in the cellar top mud is controlled to ≤11%, the growth of mold on the surface of the cellar top mud can be inhibited.
[0066] Table 6 Effect of loess water content on the growth of bacteria in cellar top mud serial number Loess moisture content / % Bacteria growth E1 15 +++ E2 13 +++ E3 12 ++ E4 11 - E5 10 - E6 9 - E7 8 - Note: + represents bacterial growth, - represents no bacterial growth.
[0067] In summary, the growth of mold can be inhibited under any one of the following conditions: keeping the cellar top mud in a sealed state, controlling the ratio of the cellar top mud thickness to the fermentation container height to ≥10%, or exposing the loess in the cellar top mud to the sun (controlling the water content of the loess in the cellar top mud to ≤11%).
[0068] Example 4: Workshop Application 1. Test materials Old mud (i.e. mud from the cellar top that has undergone the previous round of fermentation), loess, and mash are the materials used in the production workshop.
[0069] 2. Test methods After the loess is exposed to the sun (water content is 11%), it is mixed with old mud in a ratio of 1:2 to make new mud (cellar top mud) with a water content of 35%, which is covered on the mash. The thickness of the cellar top mud is 25cm, and the cellar height is 250cm (the thickness of the cellar top mud and the cellar height account for 10%). Then, a layer of plastic cloth is covered on the surface and strictly sealed. If a hole needs to be inserted during fermentation, the hole needs to be pulled out and the hole is covered with the cellar top mud. The thickness of the cellar top mud in the control group is 25cm, and there is no need to cover the cellar top mud if there is a hole during the fermentation. The growth of bacteria in the cellar top mud was observed during the fermentation period.
[0070] 3. Test results Cellar Figure 5 As shown, no bacteria grew in the test group during the entire fermentation period, while bacteria grew in the pores of the test group during the fermentation period. The results confirmed that the growth of mold can be inhibited by keeping the cellar top mud in a sealed state, controlling the ratio of the cellar top mud thickness to the fermentation container height to ≥10%, or exposing the loess in the cellar top mud to the sun (controlling the water content of the loess in the cellar top mud to ≤11%).
Claims
1. A cephalosporin Aspergillus, Features: The Aspergillus cejpii is named as Aspergillus cejpii JDN-1; the preservation number of the Aspergillus cejpii JDN-1 is CCTCC NO: M 20241807.
2. The Aspergillus cephalosporin according to claim 1, Features: The Aspergillus cephalosporin JDN-1 originates from the mold on the surface of the pit top mud.
3. A method for inhibiting the growth of mold on the surface of pit top mud, It is characterized in that The mold on the surface of the pit top mud is the Aspergillus cephalosporin JDN-1 described in claim 1 or 2; the inhibition method is to use at least one of the following methods to inhibit the growth of mold on the surface of the pit top mud: (1) Adjust the temperature; (2) Anaerobic; (3) pH adjustment; (4) Lethal temperature.
4. The method according to claim 3, Features: In the step (1), the temperature is adjusted to less than 10°C.
5. The method according to claim 3, Features: In the step (1), the temperature is adjusted to > 40°C.
6. The method according to claim 3, Features: In the step (3), the pH is adjusted to 3-4.
7. The method according to claim 3, Features: The lethal temperature in step (4) is 65°C.
8. Application of a method for inhibiting mold growth on the surface of cellar top mud in industrial winemaking process, Features: The mold on the surface of the pit top mud is the Aspergillus cephalosporin JDN-1 described in claim 1 or 2; the inhibition method is to use at least one of the following methods to inhibit the growth of mold on the surface of the pit top mud: (1) Anaerobic; (2) Control the ratio of the thickness of the pit top mud to the height of the fermentation container to ≥10%; (3) Expose the loess in the mud on the cellar roof to the sun and control the water content of the loess in the mud on the cellar roof.
9. The use according to claim 8, Features: In the step (3), the water content of the loess in the pit top mud is controlled to be ≤11%.