CN115005478B - Baking method for reducing flue-cured tobacco TSNAs with different maturity by using strain - Google Patents
Baking method for reducing flue-cured tobacco TSNAs with different maturity by using strain Download PDFInfo
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Abstract
The invention relates to a baking method for reducing flue-cured tobacco TSNAs with different maturity by using bacterial strains, wherein the bacterial strains used in the baking method are bacillus subtilis and trichoderma. The baking method comprises the steps of preparing bacterial solutions with OD600 value of 1 by using bacillus subtilis and trichoderma strains respectively and independently, and then uniformly spraying and inoculating the bacterial solutions on the surfaces of the harvested fresh tobacco leaves. The strain is used for the first time in the baking process of tobacco leaves with different maturity to reduce the TSNAs content in the tobacco leaves with different maturity, the total TSNAs content in the tobacco leaves with different maturity is reduced by 13.71-29.18%, wherein NNK, NNN, NAT is respectively reduced by 10.96-27.72%, 11.16-34.88% and 12.50-26.67%.
Description
Technical Field
The invention belongs to the technical field of tobacco baking, and relates to a baking method for reducing flue-cured tobacco TSNAs with different maturity by using strains. The method can open up a road for the baking method and the new technology application of the biodegradation of the flue-cured tobacco nicotine and tobacco specific nitrosamines with different maturity.
Background
Nicotine (Nicotine) has high toxicity, is permeable to biological membranes, is chemically stable and is not easily degraded, and is an important precursor of Tobacco Specific Nitrosamines (TSNAs) which are carcinogens. When nicotine reaches the brain, it causes release of epinephrine and thus craving for nicotine. Previous studies have shown that strains capable of degrading nicotine, such as Pseudomonas sp.Nic22, aspergillus (Aspergillus oryzae 112822), arthrobacter sp.M2012083, are isolated in tobacco planting soil, tobacco lamina, and tobacco waste. However, when these strains are subjected to different environments to degrade nicotine, the amount of nicotine degraded varies. Thus, conditions for degrading nicotine using microorganisms remain to be explored.
In addition, tobacco Specific Nitrosamines (TSNAs) are one of the main harmful substances affecting human health. Numerous studies have shown that TSNA is extremely low in fresh leaves and even undetectable. For this reason, most hypothesis believes that TSNA is formed by microbial reduction of nitrate in tobacco to nitrite and Nitrogen Oxides (NOX) and then interaction with tobacco alkaloids during tobacco leaf conditioning and storage. In recent 40 years, how to reduce the TSNA content in tobacco has been the subject of attack by tobacco researchers, and has become the subject of serious research by tobacco companies worldwide, and how to reduce and regulate the TSNAs content in tobacco is very important for tobacco production and development. There is currently no effective natural degradation method. The biodegradation of TSNAs by microorganisms has attracted increasing attention due to their high efficiency and simple processing methods.
Recently, degradation of nicotine by microbial treatment has been studied extensively. For example, pseudomonas convex (Pseudomonas putida) PC1, arthrobacter oxide, arthrobacter nicotinovorans and Achromobacter all showed the ability to degrade nicotine. It is speculated that the tobacco endophyte community has been adapted to use nicotine as a growth substrate, utilizing biochemical reactions to decompose such organic heterocyclic compounds nicotine. However, the method for reducing TSNAs content in tobacco leaves by using strains in the baking process is less common, and has few reports on the reduction of TSNAs content of flue-cured tobacco with different maturity. The invention provides a baking method for reducing the tobacco TSNAs with different maturity by using strains, which can open up a road for the application of the baking method and new technology for biodegrading tobacco nicotine with different maturity and tobacco specific nitrosamine, and simultaneously makes great contribution to the realization of tar reduction and harm reduction of tobacco.
Disclosure of Invention
Aiming at the defects of the prior art, based on the maturity of tobacco leaves, the baking method for reducing the TSNAs of the tobacco leaves with different maturity by utilizing strains, which can effectively reduce the TSNAs content of the tobacco leaves so as to improve the quality of the tobacco leaves, is provided.
The invention discloses a baking method for reducing flue-cured tobacco TSNAs with different maturity by using strains, which uses bacillus subtilis and trichoderma. The bacillus subtilis and the trichoderma are commercially available.
The flue-cured tobacco with different maturity in the curing method is set to have fresh tobacco leaf SPAD values of 30-40, 20-30 and 10-20 corresponding to tobacco leaves at the upper, middle and lower 3 parts of tobacco plants of undercured, moderate cured and overcured flue-cured tobacco respectively.
The flue-cured tobacco with different maturity in the baking method is set as the flue-cured tobacco with proper maturity, wherein the standard of the fresh tobacco leaves is that main veins turn white and shiny, branch veins turn green and white, leaves are green-yellow and pale yellow, and fuzz mostly falls off, and the tobacco leaves are rich in tobacco oil after being touched by hands; judging the flue-cured tobacco harvesting time of different maturity according to the proper ripe tobacco, and taking the first 15 days of the proper ripe flue-cured tobacco harvesting time as the undercooked tobacco harvesting time. Taking the last 15 days of the harvest time of the properly ripe harvested flue-cured tobacco as the harvest time of the overripe tobacco leaves.
The baking method comprises the steps of preparing bacterial solutions with OD600 value of 1 by using bacillus subtilis and trichoderma strains respectively and independently, and then uniformly spraying and inoculating the bacterial solutions on the surfaces of the collected fresh tobacco leaves.
The uniform spraying dosage of the invention is 3-5% of the weight of tobacco leaves.
The baking method comprises the steps of uniformly spraying, braiding rods, and directly baking in a converter.
The flue-cured tobacco varieties suitable for the baking method comprise cloud 87, NC102 and KRK26.
The bacterial liquid adopts two strains of bacillus subtilis and trichoderma according to the proportion of 1:1 weight ratio.
The bacterial liquid adopts two strains of bacillus subtilis and trichoderma according to the proportion of 2:1 weight ratio.
The bacterial liquid adopts two strains of bacillus subtilis and trichoderma according to the proportion of 1:2 weight ratio.
The implementation technical scheme of the invention comprises the following steps of fresh tobacco leaf harvesting, inoculation and braiding rod standard, charging standard, baking control and TSNAs degradation efficiency detection with different maturity, and the specific technical scheme is as follows:
A. harvesting, inoculating and braiding rod standard: harvesting fresh cured tobacco leaves with different maturity degrees (undercooked, proper maturity and overripeness) after topping the cured tobacco; preparing bacterial solutions with OD600 value of 1 by bacillus subtilis and trichoderma strains respectively and independently, then uniformly spraying and inoculating the bacterial solutions on the surfaces of the harvested fresh tobacco leaves, and sorting according to maturity while inoculating the strains, and then braiding rods;
B. Charging standard: filling the fresh tobacco leaves inoculated with the braided rods into a bulk curing barn according to the maturity, and gradually reducing 2-3 rods of tobacco to form dislocation and intersection;
C. baking control: the tobacco leaves with low TSNAs can be obtained after baking is completed by controlling the yellowing period, the color fixing period and the dry rib period.
Baking process for NC102 variety:
C11. and (3) yellow period control: the yellowing stage comprises a yellowing early stage, a yellowing medium stage and a yellowing later stage, wherein the yellowing early stage is that after baking and ignition are carried out, the temperature of dry balls is raised from room temperature to 34 ℃ and the temperature of wet balls is 33 ℃ for 5-6 hours, and the dry balls and the wet balls are baked for 6-8 hours; then entering a yellowing medium stage, heating dry bulb to 38 ℃ at a heating rate of 2 ℃/3 hours for 6 hours, adjusting the wet bulb temperature to 35 ℃, and baking the wet bulb for 30-35 hours until the third tobacco leaves turn into yellow 7-8 (4 curing barns) and the middle tobacco leaves turn into yellow 9 (3 curing barns); and then the tobacco leaves enter the later yellowing stage, the temperature of the dry bulb is increased to 42 ℃ at the heating rate of 1 ℃/2.5 hours, the temperature of the wet bulb is adjusted to 36 ℃, and the dry bulb is baked for 8-12 hours until the second tobacco leaves are curled with the tips (4 curing barns) and the middle tobacco leaves are curled with the tips (3 curing barns). And then the temperature of the dry bulb is increased to 43 ℃ at the heating rate of 1.5 ℃/3 hours, the temperature of the wet bulb is adjusted to 36 ℃, and the dry bulb and the wet bulb are baked for 22-25 hours until the second tobacco small winding drum (4 curing barns) and the middle tobacco leaves are slightly smaller in size (3 curing barns). Then, heating the dry bulb temperature to 45 ℃ at a heating rate of 1 ℃/2-3 hours, adjusting the wet bulb temperature to 36 ℃, and baking the dry bulb temperature for 15-20 hours until the second tobacco small winding drum (4 baking rooms) and the middle tobacco leaves appear slightly smaller winding drum (3 baking rooms);
C21. And (3) controlling a fixed color period: the color fixing period comprises a color fixing earlier stage, a color fixing middle stage and a color fixing later stage, wherein the color fixing earlier stage is to raise the temperature of dry bulb to 47 ℃ at a heating rate of 1 ℃/3 hours after the end of the yellowing later stage, adjust the temperature of wet bulb to 36 ℃, and bake the wet bulb for 20-23 hours to a small tobacco leaf winding drum at the bottom stage, and the branch pulse of tobacco leaves is full yellow; then entering a fixed color middle stage, raising the temperature of the dry bulb to 55 ℃ at a heating rate of 1.5 ℃/2 hours, adjusting the temperature of the wet bulb to 38 ℃, and stabilizing the temperature of the dry bulb and baking for 12-15 hours to a large whole furnace winding drum; then entering the final color fixing stage, and raising the dry bulb temperature to 65 ℃ at a heating rate of 0.5 ℃/1 hour, adjusting the wet bulb temperature to 38 ℃, and baking the dry bulb temperature for 10-12 hours until the main pulse of the tobacco leaves is more than one third of the main pulse of the tobacco leaves;
C31. and (3) controlling a tendon drying period: the dry rib period is to raise the temperature of the dry ball to 68 ℃ at a heating rate of 1.5 ℃/2 hours after the color fixing period is finished, adjust the temperature of the wet ball to 39 ℃, and bake the dry ball until the main pulse of the whole furnace is completely baked.
Baking process for cloud 87:
C12. and (3) yellow period control: the yellowing stage comprises a yellowing early stage, a yellowing medium stage and a yellowing later stage, wherein the yellowing early stage is that after baking and ignition are carried out, the temperature of dry balls is raised from room temperature to 35 ℃ and the temperature of wet balls is 33 ℃ for 4-5 hours, and the dry balls and the wet balls are baked for 10-12 hours; then entering a yellowing medium stage, raising the temperature of dry bulb to 38 ℃ at a heating rate of 1 ℃/2h for 6 hours, adjusting the temperature of wet bulb to 34 ℃, and baking the dry bulb for 28-32 hours until the third tobacco leaves turn into yellow 7-8 (4 curing barns) and the middle tobacco leaves turn into yellow 8 (3 curing barns); and then the tobacco leaves enter the later yellowing stage, the temperature of dry bulb is increased to 42 ℃ at the heating rate of 1 ℃/2h, the temperature of wet bulb is adjusted to 35 ℃, and the dry bulb is baked for 10-14 h until the second tobacco leaves are curled with the tips (4 curing barns) and the middle tobacco leaves are curled with the tips (3 curing barns). And then heating the dry bulb to 44 ℃ at a heating rate of 1 ℃/3 hours, regulating the wet bulb to 36 ℃, and baking the dry bulb for 20-22 hours until the third tobacco small winding drum (4 curing barns) and the middle tobacco leaves appear slightly smaller winding drum (3 curing barns). Then raising the temperature of the dry bulb to 46 ℃ at a heating rate of 2 ℃/3 hours, regulating the temperature of the wet bulb to 36 ℃, and baking the dry bulb for 12-15 hours to a second tobacco small winding drum (4 baking rooms) and a middle tobacco small winding drum (3 baking rooms) stably;
C22. And (3) controlling a fixed color period: the color fixing period comprises a color fixing earlier stage, a color fixing middle stage and a color fixing later stage, wherein the color fixing earlier stage is to raise the temperature of dry bulbs to 48 ℃ at a heating rate of 2 ℃/3h after the yellowing later stage is finished, adjust the temperature of wet bulbs to 38 ℃, and bake the dry bulbs to the tobacco leaves at the bottom for 16-18 h to obtain small tobacco leaf reels, and the branches of the tobacco leaves are all yellow; then entering a fixed color middle stage, heating the dry bulb temperature to 54 ℃ at a heating rate of 1 ℃/h, adjusting the wet bulb temperature to 39 ℃, and stabilizing the dry bulb temperature and baking the wet bulb temperature for 18-22 hours to a full-furnace large winding drum; then entering the final color fixing stage, raising the temperature of dry bulbs to 60 ℃ at a heating rate of 1 ℃/h, raising the temperature of wet bulbs to 39 ℃, and baking the dry bulbs for 10-14 hours until the main pulse of tobacco leaves is one third or more;
C32. and (3) controlling a tendon drying period: the dry rib period is to raise the temperature of the dry ball to 68 ℃ at a heating rate of 1 ℃/h after the final period of color fixing, adjust the temperature of the wet ball to 40 ℃, and bake the main pulse of the whole baking furnace completely.
Baking process for KRK 26:
C13. and (3) yellow period control: the yellowing stage comprises a yellowing early stage, a yellowing medium stage and a yellowing later stage, wherein the yellowing early stage is that after baking and ignition are carried out, the temperature of dry balls is raised from room temperature to 34 ℃ and the temperature of wet balls is 32 ℃ for 6-7 hours, and the dry balls and the wet balls are baked for 10-14 hours; then entering a yellowing medium stage, raising the temperature of dry bulb to 37 ℃ at a heating rate of 3 ℃/4h for 4h, adjusting the temperature of wet bulb to 33 ℃, and baking the dry bulb to stable dry bulb temperature for 16-18 h until the third tobacco leaves turn yellow 7-8 (4 baking rooms) and the middle tobacco leaves turn yellow 8 (3 baking rooms); and then the tobacco leaves enter the later yellowing stage, the temperature of the dry bulb is increased to 40 ℃ at the heating rate of 2 ℃/3h, the temperature of the wet bulb is adjusted to 34 ℃, and the dry bulb is baked for 12-16 h until the second tobacco leaves are curled with the tips (4 curing barns) and the middle tobacco leaves are curled with the tips (3 curing barns). And then heating the dry bulb to 42 ℃ at a heating rate of 1 ℃/2 hours, adjusting the wet bulb to 35 ℃, and baking the dry bulb for 12-16 hours until the third tobacco small winding drum (4 curing barns) and the middle tobacco leaves are slightly displayed. Then raising the temperature of the dry bulb to 45 ℃ at a heating rate of 1 ℃/1.5 hours, regulating the temperature of the wet bulb to 36 ℃, and baking the dry bulb for 8-10 hours to a second tobacco small winding drum (4 baking rooms) and a middle tobacco small winding drum (3 baking rooms) stably;
C23. And (3) controlling a fixed color period: the color fixing period comprises a color fixing earlier stage, a color fixing middle stage and a color fixing later stage, wherein the color fixing earlier stage is to raise the temperature of dry bulbs to 48 ℃ at a heating rate of 1 ℃/2h after the yellowing later stage is finished, adjust the temperature of wet bulbs to 37 ℃, and bake the dry bulbs to the tobacco leaves at the bottom for 18-22 h, so that the branches of the tobacco leaves are all yellow; then entering a fixation middle stage, heating the dry bulb to 54 ℃ at a heating rate of 1.5 ℃/h, adjusting the wet bulb to 38 ℃, and stabilizing the dry bulb temperature and baking the wet bulb for 10-14 hours to a full-furnace large winding drum; then entering the final color fixing stage, raising the temperature of dry bulbs to 60 ℃ at a heating rate of 1 ℃/h, raising the temperature of wet bulbs to 39 ℃, and baking the dry bulbs stably for 8-10 h until the main pulse of tobacco leaves is over one third of the main pulse of tobacco leaves;
C33. and (3) controlling a tendon drying period: the dry rib period is to raise the temperature of the dry ball to 67 ℃ at a heating rate of 1.5 ℃/h after the final period of color fixing, adjust the temperature of the wet ball to 40 ℃, and bake the main pulse of the whole baking furnace completely and stably.
TSNAs degradation efficiency detection: the total TSNAs content of the tobacco which is baked and modulated after the strain is sprayed is reduced by 13.71 to 29.18 percent, wherein NNK, NNN, NAT is respectively reduced by 10.96 to 27.72 percent, 11.16 to 34.88 percent and 12.50 to 26.67 percent. The strain is considered to have the function of degrading TSNAs.
The invention has the beneficial effects that:
1. the strain is used for the first time in the baking process of tobacco leaves with different maturity to reduce the TSNAs content in the tobacco leaves with different maturity, the total TSNAs content in the tobacco leaves with different maturity is reduced by 13.71-29.18%, wherein NNK, NNN, NAT is respectively reduced by 10.96-27.72%, 11.16-34.88% and 12.50-26.67%.
2. The invention aims at specific flue-cured tobacco varieties: cloud 87, NC102 and KRK26, wherein NC102 and KRK26 are special tobacco raw materials for cloud-produced high-end cigarettes, and have important functions for guaranteeing and improving core tobacco raw materials.
3. Aiming at tobacco leaves with different maturity, the degradation rate of TSNAs is improved through reasonable matching and use of bacillus subtilis and trichoderma strains, so that the quality of the tobacco leaves is remarkably improved, and the harm of the TSNAs to human health and living environment is reduced.
The invention is further explained below in connection with specific embodiments.
Detailed Description
The invention is further illustrated, but is not limited in any way, by the following examples, and any alterations or substitutions based on the teachings of the invention are within the scope of the invention.
The implementation technical scheme of the invention comprises the following steps of fresh tobacco leaf harvesting, inoculation and braiding rod standard, charging standard, baking control and TSNAs degradation efficiency detection with different maturity, and the specific technical scheme is as follows:
A. Harvesting, inoculating and braiding rod standard: harvesting fresh cured tobacco leaves with different maturity degrees (undercooked, proper maturity and overripeness) after topping the cured tobacco; preparing bacterial solutions with an OD600 value of 1 by using bacillus subtilis and trichoderma strains (the bacillus subtilis and the trichoderma are obtained by commercial purchase) respectively and independently, then uniformly spraying and inoculating the bacterial solutions on the surfaces of the harvested fresh tobacco leaves, and braiding rods after classifying according to maturity while inoculating the strains;
B. charging standard: filling the fresh tobacco leaves inoculated with the braided rods into a bulk curing barn according to the maturity, and gradually reducing 2-3 rods of tobacco to form dislocation and intersection;
C. baking control: the tobacco leaves with low TSNAs can be obtained after baking is completed by controlling the yellowing period, the color fixing period and the dry rib period.
Baking process for NC102 variety:
C11. and (3) yellow period control: the yellowing stage comprises a yellowing early stage, a yellowing medium stage and a yellowing later stage, wherein the yellowing early stage is that after baking and ignition are carried out, the temperature of dry balls is raised from room temperature to 34 ℃ and the temperature of wet balls is 33 ℃ for 5-6 hours, and the dry balls and the wet balls are baked for 6-8 hours; then entering a yellowing medium stage, heating dry bulb to 38 ℃ at a heating rate of 2 ℃/3 hours for 6 hours, adjusting the wet bulb temperature to 35 ℃, and baking the wet bulb for 30-35 hours until the third tobacco leaves turn into yellow 7-8 (4 curing barns) and the middle tobacco leaves turn into yellow 9 (3 curing barns); and then the tobacco leaves enter the later yellowing stage, the temperature of the dry bulb is increased to 42 ℃ at the heating rate of 1 ℃/2.5 hours, the temperature of the wet bulb is adjusted to 36 ℃, and the dry bulb is baked for 8-12 hours until the second tobacco leaves are curled with the tips (4 curing barns) and the middle tobacco leaves are curled with the tips (3 curing barns). And then the temperature of the dry bulb is increased to 43 ℃ at the heating rate of 1.5 ℃/3 hours, the temperature of the wet bulb is adjusted to 36 ℃, and the dry bulb and the wet bulb are baked for 22-25 hours until the second tobacco small winding drum (4 curing barns) and the middle tobacco leaves are slightly smaller in size (3 curing barns). Then, heating the dry bulb temperature to 45 ℃ at a heating rate of 1 ℃/2-3 hours, adjusting the wet bulb temperature to 36 ℃, and baking the dry bulb temperature for 15-20 hours until the second tobacco small winding drum (4 baking rooms) and the middle tobacco leaves appear slightly smaller winding drum (3 baking rooms);
C21. And (3) controlling a fixed color period: the color fixing period comprises a color fixing earlier stage, a color fixing middle stage and a color fixing later stage, wherein the color fixing earlier stage is to raise the temperature of dry bulb to 47 ℃ at a heating rate of 1 ℃/3 hours after the end of the yellowing later stage, adjust the temperature of wet bulb to 36 ℃, and bake the wet bulb for 20-23 hours to a small tobacco leaf winding drum at the bottom stage, and the branch pulse of tobacco leaves is full yellow; then entering a fixed color middle stage, raising the temperature of the dry bulb to 55 ℃ at a heating rate of 1.5 ℃/2 hours, adjusting the temperature of the wet bulb to 38 ℃, and stabilizing the temperature of the dry bulb and baking for 12-15 hours to a large whole furnace winding drum; then entering the final color fixing stage, and raising the dry bulb temperature to 65 ℃ at a heating rate of 0.5 ℃/1 hour, adjusting the wet bulb temperature to 38 ℃, and baking the dry bulb temperature for 10-12 hours until the main pulse of the tobacco leaves is more than one third of the main pulse of the tobacco leaves;
C31. and (3) controlling a tendon drying period: the dry rib period is to raise the temperature of the dry ball to 68 ℃ at a heating rate of 1.5 ℃/2 hours after the color fixing period is finished, adjust the temperature of the wet ball to 39 ℃, and bake the dry ball until the main pulse of the whole furnace is completely baked.
Baking process for cloud 87:
C12. and (3) yellow period control: the yellowing stage comprises a yellowing early stage, a yellowing medium stage and a yellowing later stage, wherein the yellowing early stage is that after baking and ignition are carried out, the temperature of dry balls is raised from room temperature to 35 ℃ and the temperature of wet balls is 33 ℃ for 4-5 hours, and the dry balls and the wet balls are baked for 10-12 hours; then entering a yellowing medium stage, raising the temperature of dry bulb to 38 ℃ at a heating rate of 1 ℃/2h for 6 hours, adjusting the temperature of wet bulb to 34 ℃, and baking the dry bulb for 28-32 hours until the third tobacco leaves turn into yellow 7-8 (4 curing barns) and the middle tobacco leaves turn into yellow 8 (3 curing barns); and then the tobacco leaves enter the later yellowing stage, the temperature of dry bulb is increased to 42 ℃ at the heating rate of 1 ℃/2h, the temperature of wet bulb is adjusted to 35 ℃, and the dry bulb is baked for 10-14 h until the second tobacco leaves are curled with the tips (4 curing barns) and the middle tobacco leaves are curled with the tips (3 curing barns). And then heating the dry bulb to 44 ℃ at a heating rate of 1 ℃/3 hours, regulating the wet bulb to 36 ℃, and baking the dry bulb for 20-22 hours until the third tobacco small winding drum (4 curing barns) and the middle tobacco leaves appear slightly smaller winding drum (3 curing barns). Then raising the temperature of the dry bulb to 46 ℃ at a heating rate of 2 ℃/3 hours, regulating the temperature of the wet bulb to 36 ℃, and baking the dry bulb for 12-15 hours to a second tobacco small winding drum (4 baking rooms) and a middle tobacco small winding drum (3 baking rooms) stably;
C22. And (3) controlling a fixed color period: the color fixing period comprises a color fixing earlier stage, a color fixing middle stage and a color fixing later stage, wherein the color fixing earlier stage is to raise the temperature of dry bulbs to 48 ℃ at a heating rate of 2 ℃/3h after the yellowing later stage is finished, adjust the temperature of wet bulbs to 38 ℃, and bake the dry bulbs to the tobacco leaves at the bottom for 16-18 h to obtain small tobacco leaf reels, and the branches of the tobacco leaves are all yellow; then entering a fixed color middle stage, heating the dry bulb temperature to 54 ℃ at a heating rate of 1 ℃/h, adjusting the wet bulb temperature to 39 ℃, and stabilizing the dry bulb temperature and baking the wet bulb temperature for 18-22 hours to a full-furnace large winding drum; then entering the final color fixing stage, raising the temperature of dry bulbs to 60 ℃ at a heating rate of 1 ℃/h, raising the temperature of wet bulbs to 39 ℃, and baking the dry bulbs for 10-14 hours until the main pulse of tobacco leaves is one third or more;
C32. and (3) controlling a tendon drying period: the dry rib period is to raise the temperature of the dry ball to 68 ℃ at a heating rate of 1 ℃/h after the final period of color fixing, adjust the temperature of the wet ball to 40 ℃, and bake the main pulse of the whole baking furnace completely.
Baking process for KRK 26:
C13. and (3) yellow period control: the yellowing stage comprises a yellowing early stage, a yellowing medium stage and a yellowing later stage, wherein the yellowing early stage is that after baking and ignition are carried out, the temperature of dry balls is raised from room temperature to 34 ℃ and the temperature of wet balls is 32 ℃ for 6-7 hours, and the dry balls and the wet balls are baked for 10-14 hours; then entering a yellowing medium stage, raising the temperature of dry bulb to 37 ℃ at a heating rate of 3 ℃/4h for 4h, adjusting the temperature of wet bulb to 33 ℃, and baking the dry bulb to stable dry bulb temperature for 16-18 h until the third tobacco leaves turn yellow 7-8 (4 baking rooms) and the middle tobacco leaves turn yellow 8 (3 baking rooms); and then the tobacco leaves enter the later yellowing stage, the temperature of the dry bulb is increased to 40 ℃ at the heating rate of 2 ℃/3h, the temperature of the wet bulb is adjusted to 34 ℃, and the dry bulb is baked for 12-16 h until the second tobacco leaves are curled with the tips (4 curing barns) and the middle tobacco leaves are curled with the tips (3 curing barns). And then heating the dry bulb to 42 ℃ at a heating rate of 1 ℃/2 hours, adjusting the wet bulb to 35 ℃, and baking the dry bulb for 12-16 hours until the third tobacco small winding drum (4 curing barns) and the middle tobacco leaves are slightly displayed. Then raising the temperature of the dry bulb to 45 ℃ at a heating rate of 1 ℃/1.5 hours, regulating the temperature of the wet bulb to 36 ℃, and baking the dry bulb for 8-10 hours to a second tobacco small winding drum (4 baking rooms) and a middle tobacco small winding drum (3 baking rooms) stably;
C23. And (3) controlling a fixed color period: the color fixing period comprises a color fixing earlier stage, a color fixing middle stage and a color fixing later stage, wherein the color fixing earlier stage is to raise the temperature of dry bulbs to 48 ℃ at a heating rate of 1 ℃/2h after the yellowing later stage is finished, adjust the temperature of wet bulbs to 37 ℃, and bake the dry bulbs to the tobacco leaves at the bottom for 18-22 h, so that the branches of the tobacco leaves are all yellow; then entering a fixation middle stage, heating the dry bulb to 54 ℃ at a heating rate of 1.5 ℃/h, adjusting the wet bulb to 38 ℃, and stabilizing the dry bulb temperature and baking the wet bulb for 10-14 hours to a full-furnace large winding drum; then entering the final color fixing stage, raising the temperature of dry bulbs to 60 ℃ at a heating rate of 1 ℃/h, raising the temperature of wet bulbs to 39 ℃, and baking the dry bulbs stably for 8-10 h until the main pulse of tobacco leaves is over one third of the main pulse of tobacco leaves;
C33. and (3) controlling a tendon drying period: the dry rib period is to raise the temperature of the dry ball to 67 ℃ at a heating rate of 1.5 ℃/h after the final period of color fixing, adjust the temperature of the wet ball to 40 ℃, and bake the main pulse of the whole baking furnace completely and stably.
TSNAs degradation efficiency detection: the total TSNAs content of the tobacco which is baked and modulated after the strain is sprayed is reduced by 13.71 to 29.18 percent, wherein NNK, NNN, NAT is respectively reduced by 10.96 to 27.72 percent, 11.16 to 34.88 percent and 12.50 to 26.67 percent. The strain is considered to have the function of degrading TSNAs.
The amount of the strain solution with the OD600 value of 1 sprayed uniformly on the tobacco leaves in the step A is 3-5% of the weight of the tobacco leaves.
And B, uniformly spraying bacterial liquid on the surfaces of the tobacco leaves in the step A, and then directly baking in a converter.
The tobacco plants of the underripe, moderate ripe and overripe tobacco in the step A have the corresponding fresh tobacco leaf SPAD values of 31.4-35.1, 27.9-28.6 and 13.2-20.3 respectively, the standard of the fresh tobacco leaves suitable for ripe is that main pulse becomes white and shiny, branch pulse becomes green and white, leaves are green and yellow, light yellow and hair mostly fall off, tobacco leaves rich in tobacco oil are touched by hands, tobacco harvesting time of different ripe tobacco is judged according to the moderate ripe tobacco leaves, and the first 15 days of the moderate ripe tobacco harvesting time are taken as underripe tobacco harvesting time. Taking the last 15 days of the harvest time of the properly ripe harvested flue-cured tobacco as the harvest time of the overripe tobacco leaves.
The flue-cured tobacco varieties in the step A comprise cloud 87, NC102 and KRK26.
The tobacco braiding amount of the tobacco leaves in the step A is 90-110 pieces/rod;
the smoke loading amount of the bulk curing barn in the step B is 540 to 580 rods/furnace, and the smoke loading density is 40 to over45kg/m 3 And (5) filling cigarettes to more than 95% of the volume of the curing barn.
And C11, C21 and C31 are respectively the yellowing stage control, the color fixing stage control and the dry reinforcement stage control of the NC102 variety tobacco leaves with different maturity.
And C12, C22 and C32 are respectively the yellowing stage control, the fixed color stage control and the dry rib stage control of the tobacco leaves of KRK26 varieties with different maturity.
The bulk curing barn is an airflow descending bulk curing barn.
Example 1 Effect of the curing Process of the invention on TSNAs content in NC102 tobacco leaves of different maturity
S100: NC102 flue-cured fresh tobacco leaves with different maturity degrees are collected in Jiuxi town in Jiangchuan area of Yuxi city of Yunnan province, the mature fresh tobacco leaves have the characteristics of white and shiny main pulse, green and yellow branches, and most of fuzz falls off, and abundant tobacco oil is obtained after the hand touch. The cured tobacco leaves are picked up as undercooked tobacco leaves 15 days before the cured tobacco leaves are picked up. The cured tobacco leaves are picked up as overcured tobacco leaves 15 days after the cured tobacco leaves are picked up.
S200: sorting the collected fresh tobacco leaves according to the tobacco braiding amount of 90-110 pieces/rod.
S300: and uniformly spraying bacterial liquid with an OD600 value of 1 on the surface of the tobacco leaves with the braided rods according to the amount of 3-5% of the weight of the tobacco leaves.
S400: the inoculated tobacco leaves with the braided rods are put into a furnace according to the tobacco loading amount of 540 to 580 rods/furnace of 40 to 45kg/m 3 Is filled into the air flow descending type bulk curing barn to more than 95 percent of the volume.
S500: and baking the converted tobacco leaves, and obtaining the tobacco leaves with low TSNAs after a yellowing period, a color fixing period and a tendon drying period.
The bacterial liquid in S300 is prepared by bacillus subtilis and trichoderma strains respectively. Wherein:
t0 is clear water blank control liquid without inoculating the strain;
the T1 bacterial liquid is prepared by bacillus subtilis strain;
the T2 bacterial liquid is prepared by trichoderma strains;
the T3 bacterial liquid adopts two strains of bacillus subtilis and trichoderma according to the ratio of 1:1, mixing and preparing the materials according to the weight ratio;
t4 adopts two strains of bacillus subtilis and trichoderma according to the ratio of 2:1, mixing and preparing the materials according to the weight ratio;
t5 adopts two strains of bacillus subtilis and trichoderma according to the ratio of 1:2 weight ratio.
The yellowing stage in S500 includes a yellowing early stage, a yellowing middle stage and a yellowing later stage, wherein the yellowing early stage is that after baking and ignition are carried out, the temperature of dry balls is raised from room temperature to 34 ℃ and the temperature of wet balls is 33 ℃ for 5-6 hours, and the temperature of dry balls and wet balls is stabilized and baked for 6-8 hours; then entering a yellowing medium stage, heating dry bulb to 38 ℃ at a heating rate of 2 ℃/3 hours for 6 hours, adjusting the wet bulb temperature to 35 ℃, and baking the wet bulb for 30-35 hours until the third tobacco leaves turn into yellow 7-8 (4 curing barns) and the middle tobacco leaves turn into yellow 9 (3 curing barns); and then the tobacco leaves enter the later yellowing stage, the temperature of the dry bulb is increased to 42 ℃ at the heating rate of 1 ℃/2.5 hours, the temperature of the wet bulb is adjusted to 36 ℃, and the dry bulb is baked for 8-12 hours until the second tobacco leaves are curled with the tips (4 curing barns) and the middle tobacco leaves are curled with the tips (3 curing barns). And then the temperature of the dry bulb is increased to 43 ℃ at the heating rate of 1.5 ℃/3 hours, the temperature of the wet bulb is adjusted to 36 ℃, and the dry bulb and the wet bulb are baked for 22-25 hours until the second tobacco small winding drum (4 curing barns) and the middle tobacco leaves are slightly smaller in size (3 curing barns). Then, heating the dry bulb temperature to 45 ℃ at a heating rate of 1 ℃/2-3 hours, adjusting the wet bulb temperature to 36 ℃, and baking the dry bulb temperature for 15-20 hours until the second tobacco small winding drum (4 baking rooms) and the middle tobacco leaves appear slightly smaller winding drum (3 baking rooms); the color fixing period comprises a color fixing earlier stage, a color fixing middle stage and a color fixing later stage, wherein the color fixing earlier stage is to raise the temperature of dry bulb to 47 ℃ at a heating rate of 1 ℃/3 hours after the end of the yellowing later stage, adjust the temperature of wet bulb to 36 ℃, and bake the wet bulb for 20-23 hours to a small tobacco leaf winding drum at the bottom stage, and the branch pulse of tobacco leaves is full yellow; then entering a fixed color middle stage, raising the temperature of the dry bulb to 55 ℃ at a heating rate of 1.5 ℃/2 hours, adjusting the temperature of the wet bulb to 38 ℃, and stabilizing the temperature of the dry bulb and baking for 12-15 hours to a large whole furnace winding drum; then entering the final color fixing stage, and raising the dry bulb temperature to 65 ℃ at a heating rate of 0.5 ℃/1 hour, adjusting the wet bulb temperature to 38 ℃, and baking the dry bulb temperature for 10-12 hours until the main pulse of the tobacco leaves is more than one third of the main pulse of the tobacco leaves; the dry rib period is to raise the temperature of the dry ball to 68 ℃ at a heating rate of 1.5 ℃/2 hours after the color fixing period is finished, adjust the temperature of the wet ball to 39 ℃, and bake the dry ball until the main pulse of the whole furnace is completely baked.
TABLE 1 comparison of TSNAs content of cured tobacco of NC102 cured tobacco variety of different maturity
As can be seen from Table 1, the application of the bacterial liquid treatment can reduce the TSNAs content of tobacco leaves with different maturity of NC 102. The undercooked tobacco leaves are reduced by 16.24 to 27.65 percent under the condition of inoculating strains, wherein the reduction amplitude of the T5 treatment group is the largest; under the condition of inoculating the strain, the TSNAs of the suitable tobacco leaves is reduced to 18.46-27.70%, wherein the reduction amplitude is the largest in the T4 treatment; under the condition of inoculating the strain, the TSNAs of the overripe tobacco leaves is reduced to 15.63-29.11%, wherein the reduction amplitude is the largest in the T5 treatment; therefore, the inoculation strain can obviously reduce the TSNAs content of the flue-cured tobacco, and the effect of the inoculation strain is different in flue-cured tobacco with different maturity, and the effect of combining the two strains is more obvious.
Example 2 Effect of the seed culture of the invention on the Activity of different maturity tobacco seed cultures of NC102 after baking
According to example 1, the number of viable bacteria and NO of the bacterial species of the unit leaves of tobacco leaves of different maturity at this time were determined at the end of the early stage of bake yellowing and at the end of the late stage of yellowing 2 - The content is as follows.
TABLE 2 effective viable count and NO per unit area of tobacco leaves 2 - Content comparison
As can be seen from Table 2, after the strain treatment, the cured tobacco leaves with different maturity have different effective viable counts per unit area, and the precursor for eliminating nitrosamine NO 2 - Also the ability of the species to be applied in combination to enhance the removal of nitrosamine precursor NO from tobacco 2 - And the survival rate of the strain is also higher.
Example 3 Effect of the curing Process of the invention on TSNAs content in cloud 87 tobacco leaves of different maturity
S100: the cloud 87 flue-cured fresh tobacco leaves with different maturity in the Maitreya city of Yunnan province have the characteristics that main veins become white and shiny, branch veins become green and white, the leaves are green and yellow and pale yellow, fuzz mostly falls off, and abundant tobacco tar exists after the hands touch. The cured tobacco leaves are picked up as undercooked tobacco leaves 15 days before the cured tobacco leaves are picked up. The cured tobacco leaves are picked up as overcured tobacco leaves 15 days after the cured tobacco leaves are picked up.
S200: sorting the collected fresh tobacco leaves according to the tobacco braiding amount of 90-110 pieces/rod.
S300: and uniformly spraying bacterial liquid with an OD600 value of 1 on the surface of the tobacco leaves with the braided rods according to the amount of 3-5% of the weight of the tobacco leaves.
S400: the inoculated tobacco leaves with the braided rods are put into a furnace according to the tobacco loading amount of 540 to 580 rods/furnace of 40 to 45kg/m 3 Is filled into the air flow descending type bulk curing barn to more than 95 percent of the volume.
S500: and baking the converted tobacco leaves, and obtaining the tobacco leaves with low TSNAs after a yellowing period, a color fixing period and a tendon drying period.
The bacterial liquid in S300 is prepared by bacillus subtilis and trichoderma strains respectively. Wherein:
t0 is clear water blank control liquid without inoculating the strain;
the T1 bacterial liquid is prepared by bacillus subtilis strain;
the T2 bacterial liquid is prepared by trichoderma strains;
the T3 bacterial liquid adopts two strains of bacillus subtilis and trichoderma according to the ratio of 1:1, mixing and preparing the materials according to the weight ratio;
t4 adopts two strains of bacillus subtilis and trichoderma according to the ratio of 2:1, mixing and preparing the materials according to the weight ratio;
t5 adopts two strains of bacillus subtilis and trichoderma according to the ratio of 1:2, mixing and preparing the materials according to the weight ratio; .
The yellowing stage in S500 includes a yellowing early stage, a yellowing middle stage and a yellowing later stage, wherein the yellowing early stage is that after the baking is started and ignited, the temperature of dry balls is raised to 35 ℃ from room temperature, the temperature of wet balls is 33 ℃, and the dry balls and the wet balls are stably baked for 10-12 hours; then entering a yellowing medium stage, raising the temperature of dry bulb to 38 ℃ at a heating rate of 1 ℃/2h for 6 hours, adjusting the temperature of wet bulb to 34 ℃, and baking the dry bulb for 28-32 hours until the third tobacco leaves turn into yellow 7-8 (4 curing barns) and the middle tobacco leaves turn into yellow 8 (3 curing barns); and then the tobacco leaves enter the later yellowing stage, the temperature of dry bulb is increased to 42 ℃ at the heating rate of 1 ℃/2h, the temperature of wet bulb is adjusted to 35 ℃, and the dry bulb is baked for 10-14 h until the second tobacco leaves are curled with the tips (4 curing barns) and the middle tobacco leaves are curled with the tips (3 curing barns). And then heating the dry bulb to 44 ℃ at a heating rate of 1 ℃/3 hours, regulating the wet bulb to 36 ℃, and baking the dry bulb for 20-22 hours until the third tobacco small winding drum (4 curing barns) and the middle tobacco leaves appear slightly smaller winding drum (3 curing barns). Then raising the temperature of the dry bulb to 46 ℃ at a heating rate of 2 ℃/3 hours, regulating the temperature of the wet bulb to 36 ℃, and baking the dry bulb for 12-15 hours to a second tobacco small winding drum (4 baking rooms) and a middle tobacco small winding drum (3 baking rooms) stably; the color fixing period comprises a color fixing earlier stage, a color fixing middle stage and a color fixing later stage, wherein the color fixing earlier stage is to raise the temperature of dry bulbs to 48 ℃ at a heating rate of 2 ℃/3h after the yellowing later stage is finished, adjust the temperature of wet bulbs to 38 ℃, and bake the dry bulbs to the tobacco leaves at the bottom for 16-18 h to obtain small tobacco leaf reels, and the branches of the tobacco leaves are all yellow; then entering a fixed color middle stage, heating the dry bulb temperature to 54 ℃ at a heating rate of 1 ℃/h, adjusting the wet bulb temperature to 39 ℃, and stabilizing the dry bulb temperature and baking the wet bulb temperature for 18-22 hours to a full-furnace large winding drum; then entering the final color fixing stage, raising the temperature of dry bulbs to 60 ℃ at a heating rate of 1 ℃/h, raising the temperature of wet bulbs to 39 ℃, and baking the dry bulbs for 10-14 hours until the main pulse of tobacco leaves is one third or more; the dry rib period is to raise the temperature of the dry ball to 68 ℃ at a heating rate of 1 ℃/h after the final period of color fixing, adjust the temperature of the wet ball to 40 ℃, and bake the main pulse of the whole baking furnace completely.
5 treatments were performed for each of the different maturity experiments.
TABLE 3 comparison of TSNAs content of cured tobacco of different maturity cloud 87 cured tobacco variety
As can be seen from Table 3, the application of the bacterial liquid treatment can reduce the TSNAs content of tobacco leaves with different maturity of cloud 87. Under-ripening tobacco leaves are reduced by 13.73-26.59% under the condition of inoculating strains, wherein the reduction amplitude of the T4 treatment group is the largest; under the condition of inoculating the strain, the TSNAs of the suitable tobacco leaves is reduced to 20.28-29.20%, wherein the reduction amplitude is the largest in the T5 treatment; under the condition of inoculating the strain, the TSNAs of the overripe tobacco leaves is reduced to 15.00-28.37%, wherein the reduction amplitude is the largest in the T5 treatment; therefore, the inoculation strain can obviously reduce the TSNAs content of the flue-cured tobacco, and the effect of the inoculation strain is different in flue-cured tobacco with different maturity, and the effect of combining the two strains is more obvious.
Example 4 Effect of the seed culture of the invention on the Activity of different maturity tobacco seed cultures of cloud 87 after baking
According to example 3, the number of viable bacteria and NO of the bacterial species of the unit leaves of tobacco leaves of different maturity at this time were determined at the end of the early stage of bake yellowing and at the end of the late stage of yellowing 2 - The content is as follows.
TABLE 4 effective viable count and NO per unit area of tobacco leaves 2 - Content comparison
As can be seen from Table 4, after the strain treatment, the cured tobacco leaves with different maturity have different effective viable counts per unit area, and the precursor substance NO for eliminating nitrosamine 2 - Also the ability of the species to be applied in combination to enhance the removal of nitrosamine precursor NO from tobacco 2 - And the survival rate of the strain is also higher.
EXAMPLE 5 Effect of the curing Process of the invention on TSNAs content in KRK26 tobacco leaves of different maturity
S100: KRK26 flue-cured fresh tobacco leaves with different maturity are produced in Guanzhong Zhou of Yuan province of Yunnan, the mature fresh tobacco leaves have the characteristics of white and shiny main pulse, green and pale yellow branches, most of fuzz drops off, and abundant tobacco tar is produced after hand touching. The cured tobacco leaves are picked up as undercooked tobacco leaves 15 days before the cured tobacco leaves are picked up. The cured tobacco leaves are picked up as overcured tobacco leaves 15 days after the cured tobacco leaves are picked up.
S200: sorting the collected fresh tobacco leaves according to the tobacco braiding amount of 90-110 pieces/rod.
S300: and uniformly spraying bacterial liquid with an OD600 value of 1 on the surface of the tobacco leaves with the braided rods according to the amount of 3-5% of the weight of the tobacco leaves.
S400: the inoculated tobacco leaves with the braided rods are put into a furnace according to the tobacco loading amount of 540 to 580 rods/furnace of 40 to 45kg/m 3 Is filled into the air flow descending type bulk curing barn to more than 95 percent of the volume.
S500: and baking the converted tobacco leaves, and obtaining the tobacco leaves with low TSNAs after a yellowing period, a color fixing period and a tendon drying period.
The bacterial liquid in S300 is prepared by bacillus subtilis and trichoderma strains respectively. Wherein:
t0 is clear water blank control liquid without inoculating the strain;
the T1 bacterial liquid is prepared by bacillus subtilis strain;
the T2 bacterial liquid is prepared by trichoderma strains;
the T3 bacterial liquid adopts two strains of bacillus subtilis and trichoderma according to the ratio of 1:1, mixing and preparing the materials according to the weight ratio;
t4 adopts two strains of bacillus subtilis and trichoderma according to the ratio of 2:1, mixing and preparing the materials according to the weight ratio;
t5 adopts two strains of bacillus subtilis and trichoderma according to the ratio of 1:2, mixing and preparing the materials according to the weight ratio; .
The yellowing stage in S500 includes a yellowing early stage, a yellowing middle stage and a yellowing later stage, wherein the yellowing early stage is that after the baking is started and ignited, the temperature of dry balls is raised from room temperature to 34 ℃ and the temperature of wet balls is 32 ℃ for 6-7 hours, and the dry balls and the wet balls are baked for 10-14 hours; then entering a yellowing medium stage, raising the temperature of dry bulb to 37 ℃ at a heating rate of 3 ℃/4h for 4h, adjusting the temperature of wet bulb to 33 ℃, and baking the dry bulb to stable dry bulb temperature for 16-18 h until the third tobacco leaves turn yellow 7-8 (4 baking rooms) and the middle tobacco leaves turn yellow 8 (3 baking rooms); and then the tobacco leaves enter the later yellowing stage, the temperature of the dry bulb is increased to 40 ℃ at the heating rate of 2 ℃/3h, the temperature of the wet bulb is adjusted to 34 ℃, and the dry bulb is baked for 12-16 h until the second tobacco leaves are curled with the tips (4 curing barns) and the middle tobacco leaves are curled with the tips (3 curing barns). And then heating the dry bulb to 42 ℃ at a heating rate of 1 ℃/2 hours, adjusting the wet bulb to 35 ℃, and baking the dry bulb for 12-16 hours until the third tobacco small winding drum (4 curing barns) and the middle tobacco leaves are slightly displayed. Then raising the temperature of the dry bulb to 45 ℃ at a heating rate of 1 ℃/1.5 hours, regulating the temperature of the wet bulb to 36 ℃, and baking the dry bulb for 8-10 hours to a second tobacco small winding drum (4 baking rooms) and a middle tobacco small winding drum (3 baking rooms) stably; the color fixing period comprises a color fixing earlier stage, a color fixing middle stage and a color fixing later stage, wherein the color fixing earlier stage is to raise the temperature of dry bulbs to 48 ℃ at a heating rate of 1 ℃/2h after the yellowing later stage is finished, adjust the temperature of wet bulbs to 37 ℃, and bake the dry bulbs to the tobacco leaves at the bottom for 18-22 h, so that the branches of the tobacco leaves are all yellow; then entering a fixation middle stage, heating the dry bulb to 54 ℃ at a heating rate of 1.5 ℃/h, adjusting the wet bulb to 38 ℃, and stabilizing the dry bulb temperature and baking the wet bulb for 10-14 hours to a full-furnace large winding drum; then entering the final color fixing stage, raising the temperature of dry bulbs to 60 ℃ at a heating rate of 1 ℃/h, raising the temperature of wet bulbs to 39 ℃, and baking the dry bulbs stably for 8-10 h until the main pulse of tobacco leaves is over one third of the main pulse of tobacco leaves; the dry rib period is to raise the temperature of the dry ball to 67 ℃ at a heating rate of 1.5 ℃/h after the final period of color fixing, adjust the temperature of the wet ball to 40 ℃, and bake the main pulse of the whole baking furnace completely and stably.
5 treatments were performed for each of the different maturity experiments.
TABLE 5 comparison of TSNAs content of cured tobacco after curing of KRK26 cured tobacco varieties of different maturity
As can be seen from Table 5, the application of the bacterial liquid treatment can reduce the TSNAs content of the tobacco leaves with different maturity of KRK 26. Under-ripening tobacco leaves are reduced by 11.18-26.35% under the condition of inoculating strains, wherein the reduction amplitude of the T4 treatment group is the largest; under the condition of inoculating the strain, the TSNAs of the suitable tobacco leaves is reduced to 22.00-32.16%, wherein the reduction amplitude is the largest in the T5 treatment; under the condition of inoculating the strain, the TSNAs of the overripe tobacco leaves is reduced in a range of 14.37-27.64%, wherein the reduction amplitude is the largest in the T5 treatment; therefore, the inoculation strain can obviously reduce the TSNAs content of the flue-cured tobacco, and the effect of the inoculation strain is different in flue-cured tobacco with different maturity, and the effect of combining the two strains is more obvious.
EXAMPLE 6 Effect of the seed culture of the invention on the Activity of KRK26 tobacco seed cultures at different maturity after baking
According to example 5, the number of viable bacteria of the species and NO of the unit leaf blades of tobacco leaves of different maturity at this time were determined at the end of the early stage of bake yellowing and at the end of the late stage of yellowing 2 - The content is as follows.
TABLE 6 effective viable count and NO per unit area of tobacco leaves 2 - Content comparison
As can be seen from Table 6, after the strain treatment, the cured tobacco leaves with different maturity have different effective viable counts per unit area, and the precursor substance NO for eliminating nitrosamine 2 - Also the ability of the species to be applied in combination to enhance the removal of nitrosamine precursor NO from tobacco 2 - And the survival rate of the strain is also higher.
The foregoing description is only a few specific embodiments of the present invention (since the formulation of the present invention includes numerical ranges, the embodiments are not intended to be exhaustive, and the scope of the present invention includes numerical ranges and other technical gist ranges), and the details or common knowledge of the present invention is not described in any more detail herein. It should be noted that the above embodiments do not limit the present invention in any way, and it is within the scope of the present invention for those skilled in the art to obtain the technical solution by equivalent substitution or equivalent transformation. The protection scope of the present application shall be subject to the content of the claims, and the description of the specific embodiments and the like in the specification can be used for explaining the content of the claims.
Claims (6)
1. A baking method for reducing flue-cured tobacco TSNAs with different maturity by using strains is characterized by comprising the following steps:
A. harvesting, inoculating and braiding rod standard: harvesting different maturity degrees after topping of flue-cured tobacco: undercooked, proper-ripe and overripe flue-cured fresh tobacco leaves; the used strains are bacillus subtilis and trichoderma; bacterial solutions with OD600 value of 1 are prepared separately by bacterial, and then the bacterial solutions are uniformly sprayed and inoculated on the surfaces of the collected fresh tobacco leaves, wherein the uniform spraying dosage is 3-5% of the weight of the tobacco leaves; sorting according to maturity while inoculating strains, and braiding rods;
B. Charging standard: filling the fresh tobacco leaves inoculated with the braided rods into a bulk curing barn according to the maturity;
C. baking control: the tobacco leaves with low TSNAs can be obtained after baking is completed by controlling the yellowing period, the color fixing period and the dry rib period;
the bacterial liquid adopts two strains of bacillus subtilis and trichoderma according to the ratio of 1:1 or 2:1 or 1:2, mixing and preparing the materials according to the weight ratio;
the flue-cured tobacco with different maturity in the curing method is set as the flue-cured tobacco with proper maturity, wherein the standard of the fresh tobacco leaves is that main pulse turns white and shiny, branch pulse turns green and white, leaves are green yellow and pale yellow, and fuzz mostly falls off, and tobacco oil is abundant after the tobacco leaves are touched by hands; judging flue-cured tobacco harvesting time of different maturity according to the proper ripe fresh tobacco, taking the first 15 days of the proper ripe fresh tobacco harvesting time as undercooked fresh tobacco harvesting time; taking the last 15 days of the harvest time of the proper ripe fresh tobacco leaves as the harvest time of the overripe fresh tobacco leaves;
the flue-cured tobacco variety suitable for the baking method is cloud 87;
baking process for cloud 87:
C12. and (3) yellow period control: the yellowing stage comprises a yellowing early stage, a yellowing medium stage and a yellowing later stage, wherein the yellowing early stage is that after baking and ignition are carried out, the temperature of dry balls is raised from room temperature to 35 ℃ and the temperature of wet balls is 33 ℃ for 4-5 hours, and the dry balls and the wet balls are baked for 10-12 hours; then entering a yellowing medium stage, raising the temperature of dry bulb to 38 ℃ at a heating rate of 1 ℃/2h for 6 hours, adjusting the temperature of wet bulb to 34 ℃, and baking the dry bulb for 28-32 h until the third tobacco leaves in the 4 curing barns are used for yellowing 7-8 or the middle tobacco leaves in the 3 curing barns are used for yellowing 8; then the tobacco leaves enter the later yellowing stage, the temperature of dry bulb is increased to 42 ℃ at the heating rate of 1 ℃/2h, the temperature of wet bulb is adjusted to 35 ℃, and the dry bulb is baked for 10-14 h until the second tobacco leaves in 4 curing barns are used for hooking tip and curling edges or the tobacco leaves in the middle of 3 curing barns are used for hooking tip and curling edges; then, heating the dry bulb temperature to 44 ℃ at a heating rate of 1 ℃/3 hours, adjusting the wet bulb temperature to 36 ℃, and baking the dry bulb temperature for 20-22 hours until a small tobacco roll appears in a third tobacco leaf roll of the 4 curing barns or a small tobacco leaf roll appears in a middle tobacco leaf roll of the 3 curing barns; then raising the temperature of the dry bulb to 46 ℃ at a heating rate of 2 ℃/3 hours, regulating the temperature of the wet bulb to 36 ℃, and baking the dry bulb for 12-15 hours until a second tobacco small roll in the 4 baking rooms or a tobacco small roll in the middle of the 3 baking rooms is used;
C22. And (3) controlling a fixed color period: the color fixing period comprises a color fixing earlier stage, a color fixing middle stage and a color fixing later stage, wherein the color fixing earlier stage is to raise the temperature of dry bulbs to 48 ℃ at a heating rate of 2 ℃/3h after the yellowing later stage is finished, adjust the temperature of wet bulbs to 38 ℃, and bake the dry bulbs to the tobacco leaves at the bottom for 16-18 h to obtain small tobacco leaf reels, and the branches of the tobacco leaves are all yellow; then entering a fixed color middle stage, heating the dry bulb temperature to 54 ℃ at a heating rate of 1 ℃/h, adjusting the wet bulb temperature to 39 ℃, and stabilizing the dry bulb temperature and baking the wet bulb temperature for 18-22 hours to a full-furnace large winding drum; then entering the final color fixing stage, raising the temperature of dry bulbs to 60 ℃ at a heating rate of 1 ℃/h, raising the temperature of wet bulbs to 39 ℃, and baking the dry bulbs for 10-14 hours until the main pulse of tobacco leaves is one third or more;
C32. and (3) controlling a tendon drying period: the dry rib period is to raise the temperature of the dry ball to 68 ℃ at a heating rate of 1 ℃/h after the final period of color fixing, adjust the temperature of the wet ball to 40 ℃, and bake the main pulse of the whole baking furnace completely.
2. The method for reducing TSNAs in flue-cured tobacco of different maturity by using strains according to claim 1, wherein the method for curing includes direct converter curing after braiding rods while spraying uniformly.
3. A baking method for reducing flue-cured tobacco TSNAs with different maturity by using strains is characterized by comprising the following steps:
A. harvesting, inoculating and braiding rod standard: harvesting different maturity degrees after topping of flue-cured tobacco: undercooked, proper-ripe and overripe flue-cured fresh tobacco leaves; the used strains are bacillus subtilis and trichoderma; bacterial solutions with OD600 value of 1 are prepared separately by bacterial, and then the bacterial solutions are uniformly sprayed and inoculated on the surfaces of the collected fresh tobacco leaves, wherein the uniform spraying dosage is 3-5% of the weight of the tobacco leaves; sorting according to maturity while inoculating strains, and braiding rods;
B. charging standard: filling the fresh tobacco leaves inoculated with the braided rods into a bulk curing barn according to the maturity;
C. baking control: the tobacco leaves with low TSNAs can be obtained after baking is completed by controlling the yellowing period, the color fixing period and the dry rib period;
the bacterial liquid adopts two strains of bacillus subtilis and trichoderma according to the ratio of 1:1 or 2:1 or 1:2, mixing and preparing the materials according to the weight ratio;
the flue-cured tobacco with different maturity in the curing method is set as the flue-cured tobacco with proper maturity, wherein the standard of the fresh tobacco leaves is that main pulse turns white and shiny, branch pulse turns green and white, leaves are green yellow and pale yellow, and fuzz mostly falls off, and tobacco oil is abundant after the tobacco leaves are touched by hands; judging flue-cured tobacco harvesting time of different maturity according to the proper ripe fresh tobacco, taking the first 15 days of the proper ripe fresh tobacco harvesting time as undercooked fresh tobacco harvesting time; taking the last 15 days of the harvest time of the proper ripe fresh tobacco leaves as the harvest time of the overripe fresh tobacco leaves;
The flue-cured tobacco variety suitable for the baking method is NC102;
baking process for NC102 variety:
C11. and (3) yellow period control: the yellowing stage comprises a yellowing early stage, a yellowing medium stage and a yellowing later stage, wherein the yellowing early stage is that after baking and ignition are carried out, the temperature of dry balls is raised from room temperature to 34 ℃ and the temperature of wet balls is 33 ℃ for 5-6 hours, and the dry balls and the wet balls are baked for 6-8 hours; then entering a yellowing medium stage, heating dry bulb to 38 ℃ at a heating rate of 2 ℃/3 hours for 6 hours, adjusting the wet bulb temperature to 35 ℃, and baking the wet bulb for 30-35 hours until the third tobacco leaves in the 4 curing barns are used for yellowing 7-8 or the middle tobacco leaves in the 3 curing barns are used for yellowing 9; then entering the later yellowing stage, and raising the temperature of the dry bulb to 42 ℃ at a heating rate of 1 ℃/2.5 hours, adjusting the temperature of the wet bulb to 36 ℃, and baking the dry bulb for 8-12 hours until the second tobacco leaves in the 4 curing barns are hooked with tip curled edges or the tobacco leaves in the middle of the 3 curing barns are hooked with tip curled edges; then, the temperature of the dry bulb is increased to 43 ℃ at the heating rate of 1.5 ℃/3 hours, the temperature of the wet bulb is adjusted to 36 ℃, and the dry bulb is baked for 22-25 hours until the second tobacco leaf small winding drum in 4 curing barns or the tobacco leaf in the middle of 3 curing barns appears slightly smaller winding drum; then, heating the dry bulb temperature to 45 ℃ at a heating rate of 1 ℃/2-3 hours, adjusting the wet bulb temperature to 36 ℃, and baking the dry bulb temperature for 15-20 hours until a small second tobacco roll in 4 curing barns or a small tobacco roll appears in the middle of 3 curing barns;
C21. And (3) controlling a fixed color period: the color fixing period comprises a color fixing earlier stage, a color fixing middle stage and a color fixing later stage, wherein the color fixing earlier stage is to raise the temperature of dry bulb to 47 ℃ at a heating rate of 1 ℃/3 hours after the end of the yellowing later stage, adjust the temperature of wet bulb to 36 ℃, and bake the wet bulb for 20-23 hours to a small tobacco leaf winding drum at the bottom stage, and the branch pulse of tobacco leaves is full yellow; then entering a fixed color middle stage, raising the temperature of the dry bulb to 55 ℃ at a heating rate of 1.5 ℃/2 hours, adjusting the temperature of the wet bulb to 38 ℃, and stabilizing the temperature of the dry bulb and baking for 12-15 hours to a large whole furnace winding drum; then entering the final color fixing stage, and raising the dry bulb temperature to 65 ℃ at a heating rate of 0.5 ℃/1 hour, adjusting the wet bulb temperature to 38 ℃, and baking the dry bulb temperature for 10-12 hours until the main pulse of the tobacco leaves is more than one third of the main pulse of the tobacco leaves;
C31. and (3) controlling a tendon drying period: the dry rib period is to raise the temperature of the dry ball to 68 ℃ at a heating rate of 1.5 ℃/2 hours after the color fixing period is finished, adjust the temperature of the wet ball to 39 ℃, and bake the dry ball until the main pulse of the whole furnace is completely baked.
4. A method for reducing TSNAs of flue-cured tobacco of different maturity by using a seed culture according to claim 3, wherein the method for curing comprises direct converter curing after braiding rods while uniformly spraying.
5. A baking method for reducing flue-cured tobacco TSNAs with different maturity by using strains is characterized by comprising the following steps:
A. harvesting, inoculating and braiding rod standard: harvesting different maturity degrees after topping of flue-cured tobacco: undercooked, proper-ripe and overripe flue-cured fresh tobacco leaves; the used strains are bacillus subtilis and trichoderma; bacterial solutions with OD600 value of 1 are prepared separately by bacterial, and then the bacterial solutions are uniformly sprayed and inoculated on the surfaces of the collected fresh tobacco leaves, wherein the uniform spraying dosage is 3-5% of the weight of the tobacco leaves; sorting according to maturity while inoculating strains, and braiding rods;
B. charging standard: filling the fresh tobacco leaves inoculated with the braided rods into a bulk curing barn according to the maturity;
C. baking control: the tobacco leaves with low TSNAs can be obtained after baking is completed by controlling the yellowing period, the color fixing period and the dry rib period;
the bacterial liquid adopts two strains of bacillus subtilis and trichoderma according to the ratio of 1:1 or 2:1 or 1:2, mixing and preparing the materials according to the weight ratio;
the flue-cured tobacco with different maturity in the curing method is set as the flue-cured tobacco with proper maturity, wherein the standard of the fresh tobacco leaves is that main pulse turns white and shiny, branch pulse turns green and white, leaves are green yellow and pale yellow, and fuzz mostly falls off, and tobacco oil is abundant after the tobacco leaves are touched by hands; judging flue-cured tobacco harvesting time of different maturity according to the proper ripe fresh tobacco, taking the first 15 days of the proper ripe fresh tobacco harvesting time as undercooked fresh tobacco harvesting time; taking the last 15 days of the harvest time of the proper ripe fresh tobacco leaves as the harvest time of the overripe fresh tobacco leaves;
The flue-cured tobacco variety suitable for the baking method is KRK26;
baking process for KRK 26:
C13. and (3) yellow period control: the yellowing stage comprises a yellowing early stage, a yellowing medium stage and a yellowing later stage, wherein the yellowing early stage is that after baking and ignition are carried out, the temperature of dry balls is raised from room temperature to 34 ℃ and the temperature of wet balls is 32 ℃ for 6-7 hours, and the dry balls and the wet balls are baked for 10-14 hours; then entering a yellowing medium stage, raising the temperature of dry bulb to 37 ℃ at a heating rate of 3 ℃/4h for 4h, adjusting the temperature of wet bulb to 33 ℃, and baking the dry bulb for 16-18 h until the third tobacco leaves in the 4 curing barns are used for 7-8 times of yellowing or the middle tobacco leaves in the 3 curing barns are used for 8 times of yellowing; then the tobacco leaves enter the later yellowing stage, the temperature of dry bulb is increased to 40 ℃ at the heating rate of 2 ℃/3h, the temperature of wet bulb is adjusted to 34 ℃, and the dry bulb is baked for 12-16 h until the second tobacco leaves in the 4 curing barns are used for hooking tip and curling or the tobacco leaves in the middle of the 3 curing barns are used for hooking tip and curling; then raising the temperature of the dry bulb to 42 ℃ at a heating rate of 1 ℃/2 hours, adjusting the temperature of the wet bulb to 35 ℃, and baking the dry bulb for 12-16 hours until a small tobacco roll appears in a third tobacco roll in 4 curing barns or a small tobacco roll appears in the middle tobacco roll in 3 curing barns; then raising the temperature of the dry bulb to 45 ℃ at a heating rate of 1 ℃/1.5 hours, adjusting the temperature of the wet bulb to 36 ℃, and stabilizing the temperature of the dry bulb and baking for 8-10 hours until a second tobacco leaf small roll in 4 baking rooms or a tobacco leaf small roll in the middle of 3 baking rooms is used;
C23. And (3) controlling a fixed color period: the color fixing period comprises a color fixing earlier stage, a color fixing middle stage and a color fixing later stage, wherein the color fixing earlier stage is to raise the temperature of dry bulbs to 48 ℃ at a heating rate of 1 ℃/2h after the yellowing later stage is finished, adjust the temperature of wet bulbs to 37 ℃, and bake the dry bulbs to the tobacco leaves at the bottom for 18-22 h, so that the branches of the tobacco leaves are all yellow; then entering a fixation middle stage, heating the dry bulb to 54 ℃ at a heating rate of 1.5 ℃/h, adjusting the wet bulb to 38 ℃, and stabilizing the dry bulb temperature and baking the wet bulb for 10-14 hours to a full-furnace large winding drum; then entering the final color fixing stage, raising the temperature of dry bulbs to 60 ℃ at a heating rate of 1 ℃/h, raising the temperature of wet bulbs to 39 ℃, and baking the dry bulbs stably for 8-10 h until the main pulse of tobacco leaves is over one third of the main pulse of tobacco leaves;
C33. and (3) controlling a tendon drying period: the dry rib period is to raise the temperature of the dry ball to 67 ℃ at a heating rate of 1.5 ℃/h after the final period of color fixing, adjust the temperature of the wet ball to 40 ℃, and bake the main pulse of the whole baking furnace completely and stably.
6. The method for reducing TSNAs in flue-cured tobacco of different maturity by using bacteria according to claim 5, wherein the method for curing includes direct converter curing after braiding rods while spraying uniformly.
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