Method for improving quality of reconstituted tobacco

Abstract

The invention discloses a preparation method of reconstituted tobacco. The preparation method comprises the following steps: (1) preparing tobacco raw materials; (2) sterilizing; (3) fermenting an aspergillus niger seed solution; (4) separating, and pulping the solid part; fermenting the liquid part by using Wickerhamomyces anomalus; (5) preparing tobacco paste; and (6) preparing the reconstituted tobacco. The invention further discloses the reconstituted tobacco. The preparation method provided by the invention effectively solves the problem of improving the quality of the paper-making reconstituted tobacco.

Description

Technical Field

[0001] The invention belongs to the technical field of tobacco, and in particular relates to a method for improving the quality of reconstituted tobacco leaves. Background Art

[0002] Reconstituted tobacco is a tobacco product made from tobacco raw materials such as tobacco leaf fragments, tobacco dust, cigarette ash, tobacco stems or low-grade tobacco leaves produced during the tobacco production and processing process, which is processed into properties close to natural tobacco leaves. As an important component of tobacco raw materials for cigarettes, reconstituted tobacco has obvious advantages in terms of harm reduction, tar reduction, and cost control. With the continuous improvement of cigarette structure, reconstituted tobacco, as a core component supporting the diversified design of cigarette style characteristics, has become increasingly prominent in strengthening the style of cigarette products. However, the existing tobacco raw material resources of reconstituted tobacco are difficult to match the demand for high-quality reconstituted tobacco products for the improvement of cigarette product structure. Therefore, it is necessary to explore the potential for improving the quality of reconstituted tobacco raw materials from a technical level and break through technical bottlenecks.

[0003] In recent years, the tobacco industry's medium- and long-term scientific and technological development plan has proposed using "tobacco microorganisms and biofermentation technology" to solve common problems in the industry, using cutting-edge technologies and interdisciplinary knowledge and theories, and applying knowledge and theories such as microbiome, metabolic control, and biosynthesis to the research and development of flavors and fragrances and cigarette products and to improve quality and enhance flavor, in order to further promote technological progress in my country's tobacco industry.

[0004] Aspergillus niger is one of the most widely used microorganisms in the genus Aspergillus and is an important fermentation industrial strain. Aspergillus niger is widely used in the industrial production of drugs, organic acids, proteases, etc. due to its high yield, high safety, high secretion and excellent metabolic diversity. Under natural growth conditions, Aspergillus niger secretes a large number of highly active enzymes, which effectively degrade and utilize a variety of biomacromolecules (such as starch, cellulose, hemicellulose, pectin, xylan and protein, etc.). Abnormal Wickham yeast is a yeast that produces ascospores. It is distributed in a variety of fruits and traditional wine yeasts and is a non-wine yeast widely present in the winemaking environment. This yeast can secrete a variety of glycosidases such as β-D-glucosidase, β-D-xylosidase, α-L-rhamnosidase, and has a high yield of ethyl acetate. It has a certain ability to produce aroma and esters, and is an important functional microorganism in the food fermentation process.

[0005] Therefore, in order to solve the inherent quality defects of existing tobacco raw materials for reconstituted tobacco and effectively improve the quality of reconstituted tobacco, the present invention develops a method for improving the quality of reconstituted tobacco by fermenting reconstituted tobacco raw materials with composite microorganisms in liquid state. Summary of the invention

[0006] The present invention aims at the fact that the existing tobacco raw materials for papermaking reconstituted tobacco are of low quality and cannot further meet the quality requirements of high-quality reconstituted tobacco. The present invention provides a method for improving the quality of reconstituted tobacco by fermenting the tobacco raw materials for reconstituted tobacco through composite microorganism liquid fermentation.

[0007] The technical solution of the present invention is as follows:

[0008] The first aspect of the present invention discloses a method for improving the quality of reconstituted tobacco, comprising the following steps:

[0009] (1) mixing reconstituted tobacco leaf tobacco raw materials in proportion to form reconstituted tobacco leaf raw materials;

[0010] (2) mixing the reconstituted tobacco leaf raw material with water in a weight ratio of 1:(6-8) and sterilizing the mixture;

[0011] (3) Inoculating Aspergillus niger seed liquid for the first stage of liquid fermentation for 36-48 hours, with an inoculum amount of (1-7)×(108-109) cFu / 1000 mL of fermentation liquid; fermentation temperature of 30-40° C., stirring speed of 10-30 rpm, and ventilation volume of 150-200 L / h;

[0012] (4) solid-liquid separation to obtain a solid part and a liquid part; the solid part and the added fiber are pulped separately and then mixed into a reconstituted tobacco slurry; the liquid part is subjected to a second stage of liquid fermentation, and the fermentation steps are as follows: the liquid part is inoculated with abnormal Wickham yeast seed liquid, and fermentation nutrients are added, and the fermentation is continued for 48-50 hours; the inoculation amount is (3-8)×(10 7 ~10 8 ) cFu / 1000mL fermentation liquid, fermentation temperature 25-35°C, stirring speed 5-10 rpm, ventilation volume 5-10L / h;

[0013] (5) After the second stage of liquid fermentation in step (4) is completed, the liquid portion after fermentation is subjected to membrane filtration using a 200-500 nm ceramic membrane, and then concentrated to obtain a tobacco paste;

[0014] (6) The slurry obtained in step (4) is molded into a reconstituted tobacco leaf base, the tobacco paste obtained in step (5) is coated on the leaf base, and the reconstituted tobacco leaf is obtained after drying.

[0015] Preferably, step (2) is first sterilized by high pressure and then by high temperature.

[0016] Preferably, the method for preparing the Aspergillus niger seed liquid in step (3) is: culturing in a PDA solid culture medium at 28° C. for 5 days, washing with sterile saline and preparing a bacterial suspension.

[0017] Preferably, the preparation method of the abnormal Wickham yeast seed liquid in step (4) is: fermenting in YPD medium at 30° C. and 180 r / min for 48 hours.

[0018] Preferably, the fermentation nutrient salts in step (4) are: glutamic acid (0.01-0.6) g / 1000 mL fermentation broth, sodium citrate (0.05-0.2) g / 1000 mL fermentation broth.

[0019] The second aspect of the present invention discloses reconstituted tobacco leaves prepared by the method.

[0020] Beneficial effects of the present invention:

[0021] 1. The method of the present invention sequentially uses Aspergillus niger and abnormal Wickham yeast as composite flora to ferment reconstituted tobacco raw materials. The two microorganisms act synergistically to promote the degradation of macromolecular substances in the tobacco raw materials, and use them as fermentation substrates to metabolize and produce aroma components with typical aroma characteristics.

[0022] 2. The fermentation nutrient salts involved in the method of the present invention are optimized and determined on the basis of clarifying the metabolic network and mechanism of action of abnormal Wickham yeast, and the focus is on regulating the corresponding coding genes to promote the metabolism of abnormal Wickham yeast to produce alcohol and ester flavor compounds.

[0023] 3. The reconstituted tobacco leaves obtained by the method of the present invention have significantly increased aroma components such as alcohols, esters, and lactones; at the same time, new aroma components are produced, such as TRANS-3-oxy-α-ionol, ethyl palmitate, 2-azolopyrrolidine, etc.; the aroma quality of the cigarette is significantly improved when smoking, the foreign smell and irritation are significantly reduced, and the comfort is significantly improved.

[0024] 4. The method of the present invention uses a mixture of waste materials such as tobacco fragments, tobacco dust, and tobacco stems as a tobacco raw material mixture for reconstituted tobacco leaves, and sequentially ferments them with two microorganisms, Aspergillus niger and abnormal Wickham yeast, to prepare reconstituted tobacco leaves with excellent aroma quantity and aroma quality, which are superior to tobacco products made from natural tobacco leaves; waste is utilized and the environment is protected.

[0025] Instruction Manual

[0026] Figure 1 This is the GC-MS graph of the reconstituted tobacco leaves prepared in Example 1.

[0027] Figure 2 This is the GC-MS graph of the reconstituted tobacco leaves prepared in Comparative Example 4. DETAILED DESCRIPTION

[0028] The present invention is described clearly and completely below in conjunction with the technical solutions in the embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. Other technical solutions obtained by those skilled in the art without creative work are all within the scope of protection of the present invention.

[0029] Example 1: A method for improving the quality of reconstituted tobacco leaves, comprising the following steps:

[0030] (1) mixing tobacco scraps, tobacco dust, tobacco stems and other tobacco waste materials uniformly as reconstituted tobacco leaf tobacco raw materials;

[0031] (2) mixing the reconstituted tobacco leaf raw material with water in a weight ratio of 1:6, sterilizing by high pressure for 15 min, and sterilizing again at 115° C. for 15 min;

[0032] (3) After sterilization, the Aspergillus niger seed liquid was inoculated for the first stage of liquid fermentation for 45 hours, the inoculation amount was 5×(108-109) cFu / 1000 mL of fermentation liquid, the fermentation temperature was 35°C, the stirring speed was 25 rpm, and the ventilation volume was 180 L / h; the preparation method of the Aspergillus niger seed liquid was: incubated in PDA solid culture medium at 28°C for 5 days, rinsed with sterile saline and prepared a bacterial suspension;

[0033] (4) After the first stage of fermentation, solid-liquid separation was performed to obtain a solid part and a liquid part. The solid part and the added fiber were pulped and mixed into a reconstituted tobacco slurry, and the liquid part was subjected to a second stage of liquid fermentation. The second stage of liquid fermentation of the liquid part was performed as follows: the liquid part was inoculated with abnormal Wickham yeast seed liquid, and fermentation nutrients (glutamic acid 0.3 g / 1000 mL fermentation liquid, sodium citrate 0.15 g / 1000 mL fermentation liquid) were added, and the fermentation was continued for 48 h; the inoculation amount was 4×(10 7 ~10 8 ) cFu / 1000mL fermentation liquid, fermentation temperature 30°C, stirring speed 10 rpm, ventilation volume 7L / h; the preparation method of abnormal Wickham yeast seed liquid is: fermentation in YPD medium at 30°C and 180r / min for 48h;

[0034] (5) After the second stage of liquid fermentation in step (4) is completed, the liquid portion after fermentation is subjected to membrane filtration treatment using a 300 nm ceramic membrane, and then concentrated to obtain a tobacco paste;

[0035] (6) The slurry obtained in step (4) is molded into a reconstituted tobacco leaf base, and the tobacco paste obtained in step (5) is coated on the leaf base, and the reconstituted tobacco leaf is obtained after drying.

[0036] Embodiment 2: A method for improving the quality of reconstituted tobacco leaves, comprising the following steps:

[0037] (1) Same as Example 1;

[0038] (2) mixing the reconstituted tobacco leaf raw material with water in a weight ratio of 1:8, sterilizing by high pressure for 15 min, and sterilizing again at 115° C. for 15 min;

[0039] (3) After sterilization, the Aspergillus niger seed liquid was inoculated for the first stage of liquid fermentation for 40 hours, the inoculation amount was 2×(108-109) cFu / 1000 mL of fermentation liquid, the fermentation temperature was 30°C, the stirring speed was 15 rpm, and the ventilation volume was 150 L / h; the preparation method of the Aspergillus niger seed liquid was: incubated in PDA solid culture medium at 28°C for 5 days, rinsed with sterile saline and prepared a bacterial suspension;

[0040] (4) After the first stage of fermentation, solid-liquid separation is performed to form a solid part of the tobacco raw material module and a tobacco extract. The solid part and the added fiber are pulped and mixed into a reconstituted tobacco slurry, and the tobacco extract undergoes a second stage of liquid fermentation. The second stage of liquid fermentation of the liquid part comprises the following steps: the liquid part is inoculated with abnormal Wickham yeast seed liquid, and fermentation nutrients (glutamic acid 0.02 g / 1000 mL fermentation liquid, sodium citrate 0.06 g / 1000 mL fermentation liquid) are added, and the fermentation is continued for 48 h. The inoculation amount is 1×(10 7 ~10 8 ) cFu / 1000mL fermentation liquid, fermentation temperature 26°C, stirring speed 5 rpm, ventilation volume 5L / h; the preparation method of abnormal Wickham yeast seed liquid is: fermentation in YPD medium at 30°C and 180r / min for 48h;

[0041] (5) Same as Example 1;

[0042] (6) Same as Example 1.

[0043] Comparative Example 1: Aspergillus niger was not inoculated, and the rest was the same as Example 1. The steps are as follows:

[0044] (1) Same as Example 1;

[0045] (2) Same as Example 1;

[0046] (3) solid-liquid separation is performed to obtain a solid part and a liquid part, the solid part and the added fiber are pulped separately and then mixed into a reconstituted tobacco slurry, and the liquid part is subjected to a second-stage liquid fermentation; the second-stage liquid fermentation of the liquid part comprises the following steps: the liquid part is inoculated with abnormal Wickham yeast seed liquid, and fermentation nutrients (glutamic acid 0.3 g / 1000 mL fermentation liquid, sodium citrate 0.15 g / 1000 mL fermentation liquid) are added, and the fermentation is continued for 48 h; the inoculation amount is 4×(10 7 ~10 8) cFu / 1000mL fermentation liquid, fermentation temperature 30°C, stirring speed 10 rpm, ventilation volume 7L / h; the preparation method of abnormal Wickham yeast seed liquid is: fermentation in YPD medium at 30°C and 180r / min for 48h;

[0047] (4) Same as step (5) in Example 1;

[0048] (5) Same as step (6) in Example 1.

[0049] Comparative Example 2: Abnormal Wickham yeast was not inoculated, and the rest was the same as Example 1. The steps were as follows:

[0050] (1) Same as Example 1;

[0051] (2) Same as Example 1;

[0052] (3) Same as Example 1;

[0053] (4) After the fermentation is completed, solid-liquid separation is performed to form a solid part and a liquid part of the tobacco raw material. The solid part and the added fiber are pulped separately and mixed to form a reconstituted tobacco leaf pulp; the liquid part is fermented with normal Wickham yeast;

[0054] (5) The liquid portion is filtered using a 300 nm ceramic membrane and then concentrated to obtain a tobacco paste;

[0055] (6) Same as Example 1.

[0056] Comparative Example 3: No fermentation nutrient salt was added, and the rest was the same as Example 1. The steps are as follows:

[0057] (1) Same as Example 1;

[0058] (2) Same as Example 1;

[0059] (3) Same as Example 1;

[0060] (4) After the first stage of fermentation, solid-liquid separation is performed to form a solid part and a liquid part of the tobacco raw material, the solid part and the added fiber are pulped separately and mixed into a reconstituted tobacco slurry, and the tobacco liquid part undergoes a second stage of liquid fermentation;

[0061] (5) The second stage of liquid fermentation of the liquid part is as follows: the tobacco extract is inoculated with abnormal Wickham yeast and fermented for 48 hours. The inoculation amount is 4×(10 7 ~10 8 )cFu / 1000mL fermentation liquid, fermentation temperature 30℃, stirring speed 10 rpm, ventilation volume 7L / h; no fermentation nutrient salts were added;

[0062] (6) Same as Example 1;

[0063] (7) Same as Example 1.

[0064] Comparative Example 4: No bacteria were added, and other conditions were the same as in Example 1. The steps are as follows:

[0065] (1) Same as Example 1;

[0066] (2) mixing tobacco raw material and water in a weight ratio of 1:6 and then heating and extracting at an extraction temperature of 60° C. for 30 min;

[0067] (3) After the extraction, solid-liquid separation is performed to form a solid part and a liquid part of the tobacco raw material. The solid part and the added fiber are pulped separately and mixed into a reconstituted tobacco leaf slurry, and the liquid part is concentrated to obtain a tobacco paste.

[0068] (4) The slurry obtained in step (3) is molded into a reconstituted tobacco leaf base, the obtained tobacco paste is coated on the leaf base, and the reconstituted tobacco leaf is obtained after drying.

[0069] The aroma components of the reconstituted tobacco leaves prepared in the embodiment and the comparative example were analyzed and tested. The reconstituted tobacco leaf samples were extracted with dichloromethane solvent and concentrated under reduced pressure to prepare a sample concentrate, which was further analyzed by GC-MS. The relative content of the flavor compounds was calculated based on the ratio of the peak area of ​​the flavor substance to the peak area of ​​the internal standard. At the same time, the sensory quality of the reconstituted tobacco leaves prepared in the embodiment and the comparative example was evaluated to evaluate the effect of each sample. The results are shown in Tables 1 and 2.

[0070] Table 1 Detection results of aroma components of reconstituted tobacco samples

[0071] serial number Relative content of aroma components (%) Example 1 147.25±4.37 Example 2 122.18±7.38 Comparative Example 1 110.12±5.34 Comparative Example 2 112.54±7.04 Comparative Example 3 109.27±6.31 Comparative Example 4 100.41±5.14

[0072] Table 2 Sensory quality evaluation results of reconstituted tobacco samples

[0073]

[0074] The results in Table 1 show that the sensory quality of the reconstituted tobacco leaves prepared in Example 1 is the best when smoked, followed by Example 2; the sensory qualities of Comparative Examples 1-3 are far lower than those of Example 1, and although the sensory qualities of Comparative Examples 1-3 are better than those of the reconstituted tobacco leaves in Comparative Example 4, they are difficult to achieve the effects of Examples 1 and 2. At the same time, the comparison results of the changes in the content of aroma components in Table 1 also confirm that Example 1 has the best effect; the changes in the content of aroma components in Table 1 and the comparison results of the sensory quality effect evaluation in Table 2 both confirm that the sensory quality effect of the reconstituted tobacco leaves prepared in Example 1 is the best when smoked, and the content of aroma components is the highest.

[0075] Table 3 Test results of main aroma components of reconstituted tobacco leaves prepared in Example 1 and Comparative Example 4

[0076] Chinese name Comparative Example 4 / % Example 1 / % 4-Methoxy-1-butanol 2.26 4.78 Isoamyl alcohol 3.87 7.95 2,4-Dimethylheptan-4-ol 1.74 4.37 Isosorbide-5-Nitrae Cyclohexanol 2.43 15.74 Hexaethylene glycol 2.78 9.67 2,3-Butanediol - 58.74 Cineole - 16.47 Ginseng epoxy alkynol - 9.36 Benzyl alcohol 10.78 18.65 Nonaethylene glycol 4.31 6.91 Adonit 3.16 6.54 Phenylethyl alcohol - 148.32 Furfuryl alcohol 3.67 5.92 TRANS-3-O-α-ionol - 86.34 3-Hydroxyoctadecanoic acid methyl ester - 42.97 Diethyl iminodiacetate 1.78 5.38 Octylethylene glycol mono-n-dodecyl ester 2.67 6.17 Dodecyl butyrate 1.64 4.06 Ethyl palmitate - 67.57 DL-Panthocyanolactone 10.67 18.64 Stearyl acetate - 43.97 3-(1H-Benzimidazol-3-yl)-3-hydroxy-propionate 1.93 2.13 Dihydroactin 7.24 8.34 Dihydro-3-hydroxy-4,4-dimethyl-2(3H)furanone - 37.64 Progesterone 27.34 31.93 15-crown-5-cyclohexanone - 51.67 2-Azohexanone - 101.34 4-Methoxybutan-2-one 0.91 1.63 E-5-Isopropyl-8-methyl-6,8-nonadien-2-one 6.24 7.64 2-Pyrrolidone 10.98 11.68 Rhus Tomentosa Flavonoids - 107.31

[0077] Figure 1 This is the GC-MS graph of the reconstituted tobacco leaves prepared in Example 1; Figure 2 GC-MS diagram of the reconstituted tobacco prepared by comparative example 4. Table 3 is the detection result of the aroma components of the reconstituted tobacco prepared by embodiment 1 and comparative example 4. As shown in Table 3, after the reconstituted tobacco raw material was fermented by sequentially using aspergillus niger and abnormal Wickham yeast as composite flora, the type and content of volatile aroma components were improved, and especially the aroma components of alcohols, esters and ketones were obviously improved. As shown in Table 3, new aroma components appeared after the reconstituted tobacco raw material was fermented by sequentially using aspergillus niger and abnormal Wickham yeast, such as RANS-3-oxygen-α-ionol, ethyl palmitate, Rhus flavonoids etc. This obviously improved the aroma quantity and aroma quality when the reconstituted tobacco was smoked.

[0078] The results of various embodiments and comparative examples confirm that Example 1 has the best effect, the two microorganisms Aspergillus niger and abnormal Wickham yeast can synergize well, the content of aroma components in the reconstituted tobacco leaf samples is increased and new aroma components appear; at the same time, fermentation nutrient salts play a key role in it.

[0079] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for improving the quality of reconstituted tobacco, characterized in that: The steps include: (1) mixing reconstituted tobacco leaf tobacco raw materials in proportion to form reconstituted tobacco leaf raw materials; (2) mixing the reconstituted tobacco leaf raw material with water in a weight ratio of 1:(6-8) and sterilizing the mixture; (3) Inoculating Aspergillus niger seed liquid for the first stage of liquid fermentation for 36-48 hours, with an inoculum amount of (1-7)×(108-109) cFu / 1000 mL of fermentation liquid; fermentation temperature of 30-40° C., stirring speed of 10-30 rpm, and ventilation volume of 150-200 L / h; (4) solid-liquid separation to obtain a solid part and a liquid part; the solid part and the added fiber are pulped separately and then mixed into a reconstituted tobacco slurry; the liquid part is subjected to a second stage of liquid fermentation, and the fermentation steps are as follows: the liquid part is inoculated with abnormal Wickham yeast seed liquid, and fermentation nutrients are added, and the fermentation is continued for 48-50 hours; the inoculation amount is (3-8)×(10 7 ~10 8 ) cFu / 1000mL fermentation liquid, fermentation temperature 25-35°C, stirring speed 5-10 rpm, ventilation volume 5-10L / h; (5) After the second stage of liquid fermentation in step (4) is completed, the liquid portion after fermentation is subjected to membrane filtration using a 200-500 nm ceramic membrane, and then concentrated to obtain a tobacco paste; (6) The slurry obtained in step (4) is molded into a reconstituted tobacco leaf base, the tobacco paste obtained in step (5) is coated on the leaf base, and the reconstituted tobacco leaf is obtained after drying.

2. The method according to claim 1, characterized in that Step (2) is first sterilized by high pressure and then by high temperature.

3. The method according to claim 1, characterized in that The method for preparing the Aspergillus niger seed liquid in step (3) is as follows: culturing in a PDA solid culture medium at 28° C. for 5 days, washing with sterile saline and preparing a bacterial suspension.

4. The method according to claim 1, characterized in that The method for preparing the abnormal Wickham yeast seed liquid in step (4) is: fermenting in YPD medium at 30° C. and 180 r / min for 48 hours.

5. The method according to claim 1, characterized in that The fermentation nutrient salts in step (4) are: glutamic acid (0.01-0.6) g / 1000 mL fermentation liquid, sodium citrate (0.05-0.2) g / 1000 mL fermentation liquid.

6. Reconstituted tobacco prepared according to any one of claims 1 to 5.

Images