CN111789282A - Composite preparation for improving quality of upper tobacco leaves of flue-cured tobaccos as well as preparation method and application of composite preparation - Google Patents
Composite preparation for improving quality of upper tobacco leaves of flue-cured tobaccos as well as preparation method and application of composite preparation Download PDFInfo
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- A—HUMAN NECESSITIES
- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
- A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
- A24B15/00—Chemical features or treatment of tobacco; Tobacco substitutes, e.g. in liquid form
- A24B15/18—Treatment of tobacco products or tobacco substitutes
- A24B15/20—Biochemical treatment
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
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Abstract
The invention belongs to the technical field of tobacco production, and particularly discloses a composite preparation for improving the quality of upper tobacco leaves of flue-cured tobaccos, and a preparation method and application thereof. The compound preparation comprises a first inoculum containing bacillus cereus 4-a, a second inoculum containing bacillus cereus 4-d and an enzyme preparation, wherein the preservation number of the bacillus cereus 4-a is CGMCC No.3864, and the preservation number of the bacillus cereus 4-d is CGMCC No. 5977. Compared with the control tobacco leaves which do not adopt the technology, the tobacco leaves on the upper part of the flue-cured tobacco processed by the technical scheme of the invention are beneficial to realizing more efficient conversion of macromolecular substances in the baking process, and the tobacco leaves are obviously improved in the aspects of hierarchical structure quality, sensory evaluation score, chemical component harmony and the like.
Description
Technical Field
The invention relates to the technical field of tobacco production, in particular to a composite preparation for improving the quality of upper tobacco leaves of flue-cured tobaccos, and a preparation method and application thereof.
Background
The quality of tobacco raw materials directly determines the quality and market sales of products of cigarette industry enterprises, and the industrial availability of the tobacco raw materials greatly influences the tax level of the tobacco industry. The large backlog of low-grade tobacco leaf raw materials seriously occupies the capital of cigarette industry enterprises, and the utilization rate of the tobacco leaf raw materials is urgently needed to be improved by a new technical means, so that the stock digestion is accelerated. The upper tobacco leaves of flue-cured tobaccos in China generally have the defects of insufficient maturity, unbalanced chemical components, particularly high content of nitrogen-containing compounds (such as nicotine and total protein) and the like, so that the industrial availability of the raw materials of the upper tobacco leaves is not high (the upper tobacco leaves are defined in the following that 6 to 8 tobacco leaves are downward from the top leaves after the top of a tobacco plant is cut).
The evaluation of the industrial availability of the tobacco raw materials comprises appearance quality, smoking quality, processing performance, safety and the like, so that the improvement of the existing tobacco production technology level by a biotechnology means is necessary, and microbial resources and mature commercial enzyme products widely existing in nature provide more efficient and powerful support for the direction.
In the aspect of industrial utilization of microbial resources, research accumulation and application achievements of decades have been achieved worldwide currently, and particularly in the tobacco industry, the main aspects are that microbial separation and auxiliary tobacco leaf fermentation, yeast accelerating natural alcoholization and flavoring of tobacco leaves, and microbial agents improving the tobacco planting soil environment to improve the harmony of internal chemical components of tobacco leaves. In the aspect of using enzymes, the auxiliary quality improvement directions such as the matching research of the enzymes and the essence monomers, the research of the treatment effect of a single enzyme on the chemical components and the aroma components of the tobacco leaves and the like are taken as the main points. In addition, in the technical direction of tobacco agricultural production, the harvest maturity, the water and fertilizer management level and the like are improved through field cultivation management, fine variety breeding, modulation (baking) process improvement. The research progress capable of improving the quality of the upper tobacco leaves is achieved in all the directions, but the application results are relatively dispersed, the pertinence is strong, the integration of the technical level is lacked, certain limitations are caused in the tobacco leaf production, and the difference fluctuation of varieties, production areas and years which determine the quality of tobacco leaf raw materials is difficult to deal with.
Disclosure of Invention
The invention aims to overcome the main quality defects of tobacco leaves at the upper part of flue-cured tobacco in China at present, provides a microorganism and enzyme compound preparation which can be added before the tobacco leaves are harvested, and provides a specific preparation method of the preparation and an application method in the field of tobacco production.
In order to achieve the purpose, the invention adopts the following technical scheme:
the composite preparation for improving the quality of upper tobacco leaves of flue-cured tobaccos comprises a first inoculum containing bacillus cereus 4-a, a second inoculum containing bacillus cereus 4-d and an enzyme preparation, wherein the preservation number of the bacillus cereus 4-a is CGMCC No.3864, and the preservation number of the bacillus cereus 4-d is CGMCC No. 5977.
Further, the compound preparation also comprises an inorganic salt solution auxiliary agent, wherein the inorganic salt solution auxiliary agent is prepared by uniformly mixing inorganic salt potassium citrate and sodium dihydrogen phosphate according to the proportion of 2:3 and preparing a solution with the mass fraction of 0.05-0.1%.
Further, the enzyme preparation comprises 10-12 ten thousand U/ml neutral protease, 2-3 ten thousand U/ml alpha-amylase, 20-25 ten thousand U/ml cellulase, 6-8 ten thousand U/ml pectinase and 2-3 ten thousand U/ml beta-glucanase; the neutral protease composing the enzyme preparation is calculated by volume ratio: alpha-amylase: cellulase: and (3) pectinase: beta-glucanase 8-9:50-52:1-1.2:16-17: 9-10.
Further, the cell concentrations of the first and second inoculants were 108-109One per ml.
Further, the volume ratio of the first inoculum, the second inoculum, the enzyme preparation and the inorganic salt solution adjuvant is 1-1.2:10-12:35-38: 12-15.
The invention also provides a preparation method of the composite preparation for improving the quality of tobacco leaves on the upper part of flue-cured tobacco, which comprises the following steps:
activating and culturing strains, namely respectively performing activation culture on the bacillus cereus 4-a and the bacillus cereus 4-d which are obtained by screening and separating on an activation plate culture medium;
performing amplification culture on the strains, namely performing amplification culture on single colonies of the two strains formed by the growth of the activation culture in an amplification liquid culture medium respectively to obtain bacterial suspensions;
preparing an inoculum, namely respectively collecting thalli in two bacterial suspensions, and respectively suspending the thalli in 0.85% physiological saline containing 0-0.05% tryptone to obtain a first inoculum and a second inoculum;
compounding the preparation: respectively weighing the raw materials according to the volume ratio of the first inoculum to the second inoculum to the enzyme preparation to the inorganic salt solution of 1-1.2:10-12:35-38:12-15, and uniformly mixing to obtain the composite preparation.
Further, the formula of the activation plate culture medium is as follows: 0.3% of beef extract, 0.5% of tryptone, 1% of sodium chloride, 1.5% of agar powder and the balance of distilled water, wherein the pH value of the activation plate medium is 7.2.
Further, the expanded culture step of the strain comprises inoculating the activated single colony in an expanded liquid culture medium, culturing at a constant temperature shaking table at 150rpm and 38 ℃ for more than 12 hours until the viable count reaches 106Inoculating 1ml of the bacterial liquid into the enlarged liquid culture medium again, culturing in a constant temperature shaking table at 170rpm and 38 ℃ for more than 24 hours until the viable count reaches 108-109And (4) obtaining bacterial suspension.
Further, the formula of the expanded liquid culture medium is as follows: 1% tryptone, 0.5% yeast extract, 1% sodium chloride, and the balance distilled water, and the pH of the liquid medium expanded was 7.0.
Further, in the preparation step of the inoculum, the bacterial suspensions of the two strains are respectively centrifuged for 20 minutes at 5000rpm and 4 ℃ to collect the bacterial strains, then 0.85% sterilized normal saline is used for scattering the bacterial strains and cleaning the bacterial strains, the bacterial strains are centrifuged for 10 minutes at 5000rpm and 4 ℃ for three times, and finally the collected bacterial strains are suspended in 0.85% normal saline containing 0-0.05% tryptone.
The invention also provides an application method of the composite preparation for improving the quality of the upper tobacco leaves of the flue-cured tobaccos, which is to uniformly spray the composite preparation on the front and back surfaces of the upper tobacco leaves 7 days before the tobacco leaves are harvested until visible liquid drops appear on the surfaces of the tobacco leaves.
Further, the amount of the complex preparation was 350ml/1100 nicotiana tabacum.
Further, the compound preparation is prepared by mixing the following components in a dilution ratio of 1:22 adding water to dilute and fix the volume to obtain the working solution of the compound preparation.
The invention has the beneficial effects that:
(1) the invention combines the technologies of microorganism culture, enzyme preparation screening and adjuvant compounding and adjusts the baking process, comprehensively utilizes various technologies, solves the main quality defects commonly existing in the upper tobacco leaves of the flue-cured tobaccos in China, and improves the industrial availability of the flue-cured tobaccos.
(2) Tests prove that compared with the control tobacco leaves which do not adopt the technology, the tobacco leaves on the upper part of the flue-cured tobacco processed by the technical scheme of the invention are beneficial to realizing more efficient conversion of macromolecular substances in the baking process, and are obviously improved in the aspects of tobacco leaf hierarchical structure quality, sensory evaluation score, chemical component harmony and the like.
Detailed Description
The following further illustrates the method of use and technical details of the present invention in conjunction with specific embodiments. The raw materials and the detection and analysis methods involved in the examples are briefly described as follows:
(1) the Bacillus cereus in the composite preparation is divided into two kinds, namely Bacillus cereus 4-a and Bacillus cereus 4-d, wherein the Bacillus cereus 4-a is preserved in China general microbiological culture Collection center (CGMCC) at 5/21 2010, and the preservation number is CGMCC No. 3864; the bacillus cereus 4-d is preserved in China general microbiological culture Collection center (CGMCC) at 4 months and 10 days in 2012, and the preservation number is CGMCC No. 5977. Both strains used in the examples were slant strains stored by the same company, and they were obtained from the above-mentioned depository, or other strains of the same species.
(2) The liquid enzyme product used in the invention should ensure the enzyme activity range as follows: neutral protease (10-12 ten thousand U/ml), alpha-amylase (2-3 ten thousand U/ml), cellulase (20-25 ten thousand U/ml), pectinase (6-8 ten thousand U/ml) and beta-glucanase (2-3 ten thousand U/ml).
(3) And (3) classifying the grades of the flue-cured tobacco leaves, namely classifying the samples of the whole kang tobacco leaves according to the national standard 42 grade of flue-cured tobacco leaves after the flue-cured tobacco leaves are baked, counting and comparing the weights of the tobacco leaves of all grades, summarizing the tobacco leaves into three grades of orange yellow, lemon yellow and variegated, and calculating the ratio of all grades.
(4) And (3) evaluating the sensory quality of the sample, namely wetting and shredding the processed and compared sample tobacco leaves by adopting the same process standard in a small line of a Chongqing cigarette factory, naturally airing the tobacco shreds, and rolling the tobacco shreds into cigarettes by adopting the same machine and cigarette materials according to the uniform process standard (the cigarettes are subjected to weight and suction resistance separation according to the same standard). After the sample cigarettes are balanced for 48 hours under the conditions of the temperature of 22 ℃ and the humidity of 60 percent, 9 professional evaluation committees are organized by the tobacco industry Limited liability company in Chongqing, smoking evaluation scoring is carried out on the treated and control cigarette samples according to the sensory quality evaluation method of the single material cigarettes in YC/T138-plus 1998 tobacco and tobacco product sensory evaluation method, and the sensory evaluation total score of the samples is calculated.
(5) Performing conventional chemical component detection, namely crushing the treated and contrasted sample tobacco leaves into tobacco powder by using a cyclone mill, drying the tobacco powder for 2.5 to 3 hours at the temperature of between 35 and 40 ℃, sieving the tobacco powder by using a 40-mesh sieve to obtain sample tobacco powder, and sealing and storing the sample tobacco powder; the content of chlorine, reducing sugar, total plant alkali, potassium and total nitrogen is measured by a flow analyzer according to the standard methods of YC/T162-2011 continuous flow method for measuring chlorine of tobaccos and tobacco products, YC/T159-2002 continuous flow method for measuring water-soluble sugar of tobaccos and tobacco products, YC/T160-2002-total plant alkali of tobaccos and tobacco products, YC/T217-2007 continuous flow method for measuring potassium of tobaccos and tobacco products and YC/T161-2002-total nitrogen measurement continuous flow method for measuring total nitrogen of tobaccos and tobacco products respectively, and the difference level between the processed samples and the control samples is evaluated by calculating the reference values of four chemical component coordination, namely potassium-chlorine ratio, sugar-alkali ratio, nitrogen-alkali ratio and two-sugar difference.
Examples using the above materials and technical details are listed below:
[ example 1 ]
Preparing a compound preparation: comprises the following steps of (a) carrying out,
s1: and (3) activating and culturing the strains, namely respectively activating and culturing the screened and separated bacillus cereus 4-a and bacillus cereus 4-d on an activation plate culture medium.
Specifically, in an aseptic working environment, a small amount of thalli is scraped by using a sterilizing spatula from a closed test tube slant culture medium for preserving strains, the thalli is coated on an activation plate culture medium in an aseptic culture dish by a three-zone scribing separation method, and a single colony is grown and formed, wherein the formula of the activation plate culture medium is as follows: adding 0.3% of beef extract, 0.5% of tryptone, 1% of sodium chloride, 1.5% of agar powder and distilled water to complement the volume, adjusting the pH value to 7.2, carrying out autoclaving at 121 ℃ for 10 minutes, and pouring the mixture into a sterile culture dish for solidification.
S2: and (3) performing amplification culture on the strains, namely performing amplification culture on single colonies of the two strains formed by the activation culture growth in an amplification liquid culture medium respectively to obtain bacterial suspensions.
Specifically, selecting well-grown single colony, inoculating the single colony subjected to activation culture into an enlarged liquid culture medium, and culturing at 38 deg.C for more than 12 hr at 150rpm in a constant temperature shaking table until viable count reaches 106Inoculating 1ml of the bacterial liquid into the enlarged liquid culture medium again, culturing in a constant temperature shaking table at 170rpm and 38 ℃ for more than 24 hours until the viable count reaches 108And (4) obtaining bacterial suspension. The formula of the enlarged liquid culture medium is as follows: 1% tryptone, 0.5% yeast extract, 1% sodium chloride, distilled water to complement volume, then adjusting pH value to 7.0, and autoclaving at 121 ℃ for 10 minutes.
S3: preparation of inoculum
Specifically, the bacterial suspensions of the two strains are respectively centrifuged at 5000rpm and 4 ℃ for 20 minutes to collect the bacterial strains, then 0.85% sterilized normal saline is used for scattering the bacterial strains and cleaning the bacterial strains, the bacterial strains are centrifuged at 5000rpm and 4 ℃ for 10 minutes for three times, and finally the collected bacterial strains are resuspended in 0.85% normal saline containing 0.05% tryptone, wherein the concentration of the bacterial strains in the normal saline is 108Each/ml is two kinds of inoculants which can be added on the surface of the tobacco leaf.
Selecting an enzyme preparation: the enzyme preparation comprises neutral protease (10 ten thousand U/ml, liquid), alpha-amylase (2 ten thousand U/ml, liquid), cellulase (20 ten thousand U/ml, liquid), pectinase (6 ten thousand U/ml, liquid) and beta-glucanase (2 ten thousand U/ml, liquid), and the neutral protease composing the enzyme preparation comprises the following components in volume ratio: alpha-amylase: cellulase: and (3) pectinase: β -glucanase 8:50:1:16: 9.
Selecting an inorganic salt solution auxiliary agent: the inorganic salt solution adjuvant is prepared by uniformly mixing inorganic salt potassium citrate and sodium dihydrogen phosphate according to the proportion of 2:3 and preparing a solution with the mass fraction ratio of 0.1%.
Finally, preparing 3.5L of composite preparation stock solution (liquid, can be stored for 1-2 days at 4 ℃, is preferably prepared on site) from the first inoculum, the second inoculum, the enzyme preparation solution and the inorganic salt solution auxiliary agent according to the volume ratio of 1:10:35:12, adding water for diluting to a constant volume of 1:22, and uniformly stirring to obtain the composite preparation working solution capable of being added to the upper tobacco leaves of the tobacco field of 10 mu.
Adding a compound preparation, harvesting and baking tobacco leaves: selecting 20 mu of tobacco fields with flat and connected penning water terrain and uniform growth, taking a local main cultivated variety K326 as a composite preparation adding object, removing overcast and rainy days and windy days, spraying and adding the composite preparation and a blank control to the upper tobacco leaves of the variety in cloudy or clear afternoon (avoiding sunshine insolation), uniformly spraying working solution to the front and back surfaces of the upper tobacco leaves of each flue-cured tobacco by using an electric sprayer by experienced agricultural technology until visible liquid drops appear on the surfaces of the tobacco leaves, wherein the treatment land for adding the composite preparation working solution is 10 mu (sample No. B1), and the blank control land for adding equal amount of water is 10 mu (sample No. C1) counted by about 1100 tobacco plants in 1 mu of tobacco field. 7 days after the composite preparation is added, all upper tobacco leaves are harvested once, primarily sorted to remove small leaves and diseased leaves, hung on a dense curing barn, and treated by a normal three-section six-step flue-cured tobacco curing process in Chongqing Pengshui and the like and cured by a control tobacco leaf sample. The grade structure of the flue-cured tobacco sample is shown in table 1, the sensory evaluation result of the sample cigarette is shown in table 2, and the conventional chemical component detection result and the chemical component coordination reference value of the flue-cured tobacco sample are shown in tables 3 and 4.
TABLE 1 sample grading Structure after baking
As can be seen from table 1, compared with the control sample, the orange yellow tobacco ratio of the composite preparation treatment sample is significantly increased, and the lemon yellow tobacco ratio is significantly decreased, indicating that the composite preparation treatment is beneficial to improving the hierarchical structure of the upper tobacco leaves after baking.
TABLE 2 sensory evaluation results of the sample cigarettes
As can be seen from table 2, the indexes of aroma quality, aroma amount, miscellaneous gas, irritation, penetrability, sweetness, concentration, strength and the like of the sample treated with the composite preparation are improved to different degrees compared with the control sample, and the total score is higher (increased by more than 2 scores), which indicates that the treatment with the composite preparation is beneficial to improving the sensory quality of the upper tobacco leaves after baking.
TABLE 3 test results of conventional chemical compositions of samples after baking
The results in table 3 show that compared with the control sample, the content of chlorine, reducing sugar and total sugar in the compound preparation treated sample is greatly reduced, the content of total plant alkaloid and total nitrogen is increased to a certain extent, and the content of potassium is significantly increased.
TABLE 4 reference values for chemical composition compatibility
The results in table 4 show that the potassium to chloride ratio of the composite treated sample is significantly increased compared to the control sample, indicating that the maturity and combustibility of the composite treated sample are better; the difference between the two sugars is reduced to a certain extent, which indicates that the chemical coordination of the compound preparation for treating the sample is better.
[ example 2 ]
The difference from example 1 is that, in the preparation of the complex formulation, tryptone was not contained in the physiological saline in which the inoculum 1 and the inoculum 2 had resuspended the cells; when preparing a composite preparation stock solution, the volume ratio of the first inoculum to the second inoculum to the enzyme preparation solution to the inorganic salt solution auxiliary agent is 1.2:10:38: 15; in the baking process, on the basis of a three-section six-step flue-cured tobacco baking process in Chongqing Penghui locality, the initial temperature of the yellowing stage is reduced by 4 ℃, and the temperature stabilization time of the yellowing stage is prolonged by 12 hours. The treatment sample added with the working solution of the complex formulation was numbered B2, and the blank control sample added with an equivalent amount of water was numbered C2. The grade structure of the flue-cured tobacco leaf sample is shown in table 5, the sensory evaluation result of the sample cigarette is shown in table 6, and the conventional chemical component detection result and the chemical component coordination reference value of the flue-cured tobacco leaf sample are shown in tables 7 and 8.
TABLE 5 sample grading Structure after baking
As can be seen from table 5, compared with the control sample, the orange yellow tobacco ratio of the composite preparation treatment sample is greatly increased, and the lemon yellow tobacco ratio is significantly reduced, which indicates that the improvement of the composite preparation treatment in combination with the baking process is beneficial to improving the hierarchical structure of the upper tobacco leaves after baking.
TABLE 6 sensory evaluation results of the sample cigarettes
As can be seen from table 6, the indexes of aroma quality, aroma amount, irritation, permeability, sweetness, aftertaste, concentration, strength and the like of the composite preparation treatment sample are improved to different degrees compared with the control sample, and the total score is higher (the improvement is more than 1.5 points), which indicates that the improvement of the composite preparation treatment and the baking process is favorable for improving the sensory quality of the upper tobacco leaves after baking.
TABLE 7 test results of conventional chemical compositions of samples after baking
The results in table 7 show that compared with the control sample, the content of chlorine, reducing sugar and total sugar in the compound preparation treated sample is greatly reduced, the content of total plant alkaloid and total nitrogen is increased to a certain extent, and the content of potassium is significantly increased.
TABLE 8 reference values for chemical composition compatibility
The results in table 8 show that the potassium to chloride ratio of the composite preparation treated sample is significantly increased compared to the control sample, indicating that the maturity and combustibility of the composite preparation treated sample in cooperation with the baking process improved sample are better; the difference between the two sugars is reduced to a certain extent, which indicates that the chemical coordination of the sample improved by the compound preparation treatment and the baking process is better.
[ example 3 ]
The difference from the example 1 is that in the preparation process of the composite preparation, before the thalli are collected by centrifugation, the viable count of bacterial suspensions of the two bacterial strains reaches 109Per ml; when preparing a composite preparation stock solution, the volume ratio of the first inoculum to the second inoculum to the enzyme preparation solution to the inorganic salt solution auxiliary agent is 1.1:12:35: 15; meanwhile, the concentration of the inorganic salt solution used as an auxiliary agent is 0.05 percent; neutral protease constituting the enzyme preparation: alpha-amylase: cellulase: and (3) pectinase: β -glucanase 9:52:1.2:17: 10; in the baking process, on the basis of a three-section six-step flue-cured tobacco baking process in Chongqing Penghui locality, the initial temperature of the yellowing stage is reduced by 2 ℃, and the temperature-stabilizing time of the yellowing stage is prolonged to 18 hours. The treatment sample added with the working solution of the complex formulation was numbered B3, and the blank control sample added with an equivalent amount of water was numbered C3. The grade structure of the flue-cured tobacco leaf sample is shown in table 9, the sensory evaluation result of the sample cigarette is shown in table 10, and the conventional chemical component detection result and the chemical component coordination reference value of the flue-cured tobacco leaf sample are shown in tables 11 and 12.
TABLE 9 grade Structure of baked sample
As can be seen from table 9, compared with the control sample, the ratio of orange yellow tobacco is significantly increased, the ratio of lemon yellow tobacco is reduced to a certain extent, and the ratio of mottled tobacco is also greatly reduced in the composite preparation treatment sample, which indicates that the improvement of the composite preparation treatment and the baking process is favorable for improving the hierarchical structure of the upper tobacco leaves after baking.
TABLE 10 sensory evaluation results of the sample cigarettes
As can be seen from table 10, the indexes of aroma quality, aroma amount, offensive odor, irritation, penetrability, sweetness, aftertaste, concentration, strength, and the like of the compound preparation treatment sample are improved to different degrees compared with the control sample, and the total score is higher (by more than 2.5 scores), indicating that the improvement of the compound preparation treatment in combination with the baking process is beneficial to improving the sensory quality of the upper tobacco leaves after baking.
TABLE 11 results of conventional chemical composition measurements of samples after baking
The results in table 11 show that the chlorine content of the compound preparation treated sample is reduced to a certain extent, the reducing sugar and total sugar content are greatly reduced, the total plant alkaloid content is increased to a certain extent, the potassium content is significantly increased, and the total nitrogen content is slightly reduced compared with the control sample.
TABLE 12 reference values for chemical composition compatibility
The results in table 12 show that the potassium to chloride ratio of the composite preparation treated sample is significantly increased compared to the control sample, indicating that the maturity and combustibility of the composite preparation treated sample in cooperation with the baking process improved sample are better; the difference between the two sugars is reduced to a certain extent, which indicates that the chemical coordination of the sample improved by the compound preparation treatment and the baking process is better.
The foregoing is merely an example of the present invention and common general knowledge of known specific structures and features of the embodiments is not described herein in any greater detail. It should be noted that, for those skilled in the art, without departing from the structure of the present invention, several changes and modifications can be made, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicability of the patent. The scope of the claims of the present application shall be determined by the contents of the claims, and the description of the embodiments and the like in the specification shall be used to explain the contents of the claims.
Claims (13)
1. The compound preparation for improving the quality of upper tobacco leaves of flue-cured tobaccos is characterized by comprising a first inoculum containing bacillus cereus 4-a, a second inoculum containing bacillus cereus 4-d and an enzyme preparation, wherein the preservation number of the bacillus cereus 4-a is CGMCC No.3864, and the preservation number of the bacillus cereus 4-d is CGMCC No. 5977.
2. The composite preparation for improving the quality of tobacco leaves on the upper parts of flue-cured tobaccos according to claim 1, characterized by further comprising an inorganic salt solution auxiliary agent, wherein the inorganic salt solution auxiliary agent is prepared by uniformly mixing inorganic salt potassium citrate and sodium dihydrogen phosphate according to a ratio of 2:3, and the mass fraction of the solution is 0.05% -0.1%.
3. The composite preparation for improving the quality of upper tobacco leaves of flue-cured tobacco according to claim 1, wherein the enzyme preparation comprises 10-12 ten thousand U/ml neutral protease, 2-3 ten thousand U/ml alpha-amylase, 20-25 ten thousand U/ml cellulase, 6-8 ten thousand U/ml pectinase and 2-3 ten thousand U/ml beta-glucanase; the neutral protease composing the enzyme preparation is calculated by volume ratio: alpha-amylase: cellulase: and (3) pectinase: beta-glucanase 8-9:50-52:1-1.2:16-17: 9-10.
4. According toThe composite preparation for improving the quality of upper tobacco leaves of flue-cured tobaccos according to claim 1, wherein the bacterial concentrations of the first inoculum and the second inoculum are both 108-109One per ml.
5. The composite preparation for improving the quality of upper tobacco leaves of flue-cured tobaccos according to claim 1, wherein the volume ratio of the first inoculum to the second inoculum to the enzyme preparation to the auxiliary agent of the inorganic salt solution is 1-1.2:10-12:35-38: 12-15.
6. The preparation method of the compound preparation for improving the quality of the tobacco leaves on the upper part of the flue-cured tobacco is characterized by comprising the following steps:
activating and culturing strains, namely respectively performing activation culture on the bacillus cereus 4-a and the bacillus cereus 4-d which are obtained by screening and separating on an activation plate culture medium;
performing amplification culture on the strains, namely performing amplification culture on single colonies of the two strains formed by the growth of the activation culture in an amplification liquid culture medium respectively to obtain bacterial suspensions;
preparing an inoculum, namely respectively collecting thalli in two bacterial suspensions, and respectively suspending the thalli in 0.85% physiological saline containing 0-0.05% tryptone to obtain a first inoculum and a second inoculum;
compounding the preparation: respectively weighing the raw materials according to the volume ratio of the first inoculum to the second inoculum to the enzyme preparation to the inorganic salt solution of 1-1.2:10-12:35-38:12-15, and uniformly mixing to obtain the composite preparation.
7. The method for preparing a composite preparation for improving the quality of upper tobacco leaves of flue-cured tobacco according to claim 6, wherein the formula of the activated plate culture medium is as follows: 0.3% of beef extract, 0.5% of tryptone, 1% of sodium chloride, 1.5% of agar powder and the balance of distilled water, wherein the pH value of the activated plate medium is 7.2.
8. The method for preparing a complex formulation for improving the quality of upper tobacco leaves of flue-cured tobacco according to claim 6,
the method comprises the steps of inoculating single colony subjected to activation culture into an amplification liquid culture medium, culturing for more than 12 hours in a constant temperature shaking table at the conditions of 150rpm and 38 ℃ until the viable count reaches 106Inoculating 1ml of the bacterial liquid into the enlarged liquid culture medium again, culturing in a constant temperature shaking table at 170rpm and 38 ℃ for more than 24 hours until the viable count reaches 108-109And (4) obtaining bacterial suspension.
9. The preparation method of the composite preparation for improving the quality of the upper tobacco leaves of the flue-cured tobacco according to claim 6 or 8, wherein the formula of the enlarged liquid culture medium is as follows: 1% tryptone, 0.5% yeast extract, 1% sodium chloride, and the balance distilled water, wherein the pH of the expanded liquid medium is 7.0.
10. The method for preparing a composite preparation for improving the quality of upper tobacco leaves of flue-cured tobaccos according to claim 6, wherein in the step of preparing the inoculum, bacterial suspensions of two strains are respectively centrifuged at 5000rpm and 4 ℃ for 20 minutes to collect bacterial cells, then the bacterial cells are scattered by 0.85% sterilized normal saline and then are washed, and then the bacterial cells are centrifuged at 5000rpm and 4 ℃ for 10 minutes and are repeated for three times, and finally the collected bacterial cells are suspended in 0.85% normal saline containing 0-0.05% tryptone.
11. The application method of the composite preparation according to claim 1, wherein the composite preparation is uniformly sprayed on the front and back surfaces of the upper tobacco leaves 7 days before the tobacco leaves are harvested until visible liquid drops appear on the surfaces of the tobacco leaves.
12. The use of the composition according to claim 11, wherein the amount of the complex formulation is 350ml per 1100 g of tobacco plant.
13. The use method according to claim 11 or 12, wherein the complex formulation is diluted at a dilution ratio of 1:22 adding water to dilute and fix the volume to obtain the working solution of the compound preparation.
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| CN114403496A (en) * | 2022-01-25 | 2022-04-29 | 湖北省烟草科学研究院 | Synergist for improving combustibility and ash gray level of fermented cigar tobacco leaves and application method thereof |
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