CN110786532A - Baking method for reducing TSNAs of different varieties of flue-cured tobaccos by using strains - Google Patents
Baking method for reducing TSNAs of different varieties of flue-cured tobaccos by using strains Download PDFInfo
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- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
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- A—HUMAN NECESSITIES
- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
- A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
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Abstract
The invention discloses a baking method for reducing TSNAs of different varieties of flue-cured tobaccos by utilizing strains, which comprises the steps of harvesting fresh flue-cured tobaccos with proper maturity; classifying the collected fresh tobacco leaves and then weaving the tobacco leaves into rods; preparing bacterial liquid with an OD600 value of 1 by using at least one of the bacterial strains 05-101, J45 and J54, and then uniformly spraying the bacterial liquid to be inoculated on the surface of the tobacco leaves which are already woven into the rods; loading the inoculated tobacco leaves with the braided rods into a bulk curing barn; and (4) baking the tobacco leaves after being charged into the furnace, and obtaining the tobacco leaves with low TSNAs after a yellowing period, a color fixing period and a stem drying period. According to the invention, special bacteria liquid is inoculated to the fresh tobacco leaves to be charged, and the stage change of the environment temperature and humidity in the drying chamber of the curing barn in the curing process is regulated and controlled according to the tobacco leaf variety, so that the degradation of TSNAs in the flue-cured tobacco is promoted, the purposes of regulating the TSNAs content of the tobacco leaves and improving the quality of the flue-cured tobacco are achieved, and the special bacteria liquid has very important significance for reducing the harm of the TSNAs of the tobacco to human health and living environment.
Description
Technical Field
The invention belongs to the technical field of tobacco curing, and particularly relates to a curing method for reducing TSNAs of different varieties of flue-cured tobaccos by using strains, which can effectively reduce the TSNAs content of the flue-cured tobaccos so as to improve the quality of tobacco leaves.
Background
TSNAs is a specific N-nitrosamine compound of tobacco and one of main harmful substances influencing human health. Of the 8 TSNAs identified so far, NNN, NAT, NAB and NNK are higher in 4, and NNN and NNK are both animal strong carcinogens, and the harmfulness of the NNN and NNK is proved in a high-incidence research of oral cancer of a snuff person who smells snuff for a long time without smoking. Researches show that factors such as tobacco types, tobacco structural tissues, tobacco cultivation, tobacco baking, soil microbial communities, fertilizer application amount and the like are related to the TSNAs content in tobacco. In addition, numerous studies have shown that TSNAs are present in extremely low levels in mature harvested fresh tobacco leaves, which are mainly produced and accumulated during the conditioning and baking processes. Although the prior art explores methods for reducing TSNAs in tobacco leaves from various aspects such as agricultural technology, modulation technology, biotechnology and the like, the methods for degrading TSNAs by using strains in the prior art are used for directional cleaning of pollutants, purification of drinking water and the like, and bacterial liquid is also used for air-curing and sedimentation of burley tobaccos in tobacco, the method for reducing TSNAs in tobacco leaves by using strains in the baking process is rare, and a method for reducing TSNAs content of specific varieties of flue-cured tobaccos is rarely reported.
Therefore, aiming at the characteristics of water and heat exchange in the tobacco leaves in the baking process, the TSNAs is directionally degraded in the baking process of the bulk curing barn by exploring and utilizing specific strains and combining the change of the temperature and the humidity of the environment in the drying chamber of the curing barn in the tobacco leaf baking process, so that the TSNAs has very important significance for improving the tobacco quality and reducing the harm of the TSNAs to the human health and the living environment.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides the baking method for reducing the TSNAs of different varieties of flue-cured tobaccos by using strains, which can effectively reduce the TSNAs content of the flue-cured tobaccos so as to improve the quality of the tobaccos.
The invention is realized by the following steps: the method comprises the steps of harvesting, pole weaving, inoculation, furnace charging and baking, and specifically comprises the following steps:
A. harvesting: collecting fresh flue-cured tobacco leaves with proper maturity;
B. rod weaving: classifying the collected fresh tobacco leaves and then weaving the tobacco leaves into rods;
C. inoculation: preparing bacterial liquid with an OD600 value of 1 by using at least one of the bacterial strains 05-101, J45 and J54, and then uniformly spraying the bacterial liquid to be inoculated on the surface of the tobacco leaves which are already woven into the rods;
D. charging: loading the inoculated tobacco leaves with the braided rods into a bulk curing barn;
E. baking: and (4) baking the tobacco leaves after being charged into the furnace, and obtaining the tobacco leaves with low TSNAs after a yellowing period, a color fixing period and a stem drying period.
The invention has the beneficial effects that:
1. the invention firstly uses the strain in the tobacco leaf baking process to reduce the TSNAs content in the cured tobacco, improves the quality of the cured tobacco and simultaneously proves the unique function of the strain in the tobacco leaf baking.
2. According to the invention, the specially screened strains 05-101 (with the preservation number of CCTCC NO: M2019665), J45 (with the preservation number of CCTCC NO: M2019666) and J54 (with the preservation number of CCTCC NO: M2019667) are respectively and mixedly used for reducing the TSNAs content in the flue-cured tobacco, so that the effect is obvious, the tobacco quality is improved, and the application range of the strains is widened.
3. The invention aims at specific flue-cured tobacco varieties: the tobacco is mainly planted in the prior art, and has important effects of guaranteeing and improving core tobacco raw materials.
4. Based on the self-propagation characteristics of the strains 05-101, J45 and J54, the invention provides an environment suitable for survival for the strains by adjusting temperature, hygrometer temperature and baking time in the yellowing stage and optimally combining with the baking processes in the color fixing stage and the dry stem stage, thereby improving the survival rate and the activity of inoculated strains, particularly improving the degradation rate of TSNAs by prolonging the baking time in the yellowing stage suitable for the strains to degrade the activity of TSNAs, and obviously reducing the harm of the tobacco TSNAs to the health of human bodies and living environment.
Detailed Description
The present invention is further illustrated by the following examples, but is not limited thereto in any way, and any modification or improvement based on the teaching of the present invention is within the scope of the present invention.
The invention comprises the steps of harvesting, pole weaving, inoculation, furnace charging and baking, and the specific steps are as follows:
A. harvesting: collecting fresh flue-cured tobacco leaves with proper maturity;
B. rod weaving: classifying the collected fresh tobacco leaves and then weaving the tobacco leaves into rods;
C. inoculation: preparing bacterial liquid with an OD600 value of 1 by using at least one of the bacterial strains 05-101, J45 and J54, and then uniformly spraying the bacterial liquid to be inoculated on the surface of the tobacco leaves which are already woven into the rods;
D. charging: loading the inoculated tobacco leaves with the braided rods into a bulk curing barn;
E. baking: and (4) baking the tobacco leaves after being charged into the furnace, and obtaining the tobacco leaves with low TSNAs after a yellowing period, a color fixing period and a stem drying period.
The classified name of the 05-101 strains is Alcaligenes aquatilis 05-101, which is preserved in China type culture Collection (CCTCC for short, address: China, Wuhan university Collection, zip code 430072) in 26.08.2019, and the preservation number is CCTCC number M2019665.
The classification name of the strain J45 is Bacillus altitudinis J45, which is preserved in China center for type culture Collection (CCTCC, address: China, Wuhan university Collection, zip code 430072) in 26h in 2019, and the preservation number is CCTCC number M2019666.
The classification name of the strain J54 is Bacillus altitudinis J54, which is preserved in China center for type culture Collection (CCTCC for short, address: China, Wuhan university Collection, zip code 430072) in 26.2019 at 08.month, and the preservation number is CCTCC number M2019667.
And C, uniformly spraying the strain solution with the OD600 value of 1 on the tobacco leaves, wherein the using amount of the strain solution sprayed on the tobacco leaves is 3-5% of the weight of the tobacco leaves.
And C, spraying bacterial liquid on the surface of the tobacco leaves in the step C, and directly baking in a furnace.
In the step A, SPAD values of fresh tobacco leaves corresponding to suitable maturity of tobacco leaves at the lower, middle and upper 3 parts of a flue-cured tobacco plant are 8.6-11.4, 17.9-18.6 and 13.2-20.3 respectively, or the standard of the fresh tobacco leaves suitable for maturity is that stem leaf angles are 87-93 degrees, or the standard of the fresh tobacco leaves suitable for maturity is that main veins become white and bright, branch veins turn green and white, leaves are green yellow and light yellow, most of fuzz falls off, and the flue-cured tobacco leaves are rich in tobacco tar after being touched by hands.
The flue-cured tobacco variety in the step A comprises Honghuadajinyuan, K326 and KRK 26.
And the tobacco weaving amount of the tobacco leaves in the step B is 90-110 pieces/rod.
And D, carrying out tobacco loading on the bulk curing barn by 540-580 rods/furnace, carrying out tobacco loading density by 40-45 kg/m and carrying out tobacco loading to more than 95% of the volume of the curing barn.
And E, the baking of the safflower Honghuadajinyuan tobacco leaves in the step E comprises the steps of controlling the yellowing stage, controlling the fixation stage and controlling the dry stem stage, and the steps are as follows:
e11, yellowing stage control: the yellowing period comprises an initial yellowing period, a middle yellowing period and a later yellowing period, wherein in the initial yellowing period, the temperature of dry balls in a drying chamber of a curing barn is increased to 33-34 ℃ from room temperature at a heating rate of 1 ℃/h, the temperature of wet balls is increased to 32-33 ℃ from room temperature, and then the dry and wet balls are stably baked for 16-20 h; after the early yellowing stage is finished, the dry bulb temperature is increased to 38-39 ℃ at a heating rate of 1 ℃/h in the middle yellowing stage, the wet bulb temperature is adjusted to 37-38 ℃, and then the dry bulb temperature and the wet bulb temperature are stably baked for 24-28 h; in the later yellowing stage, after the middle yellowing stage is finished, the temperature of dry balls is increased to 42-43 ℃ at the heating rate of 1 ℃/h, the temperature of wet balls is adjusted to 38-39 ℃, and then the dry and wet balls are stably baked for 16-20 h until the bottom table tobacco leaves are completely yellow;
e21, fixation period control: the ground color period comprises a color fixing initial stage and a color fixing later stage, wherein in the color fixing initial stage, after the E11 step is finished, the dry bulb temperature is increased to 47-48 ℃ at the heating rate of 0.5 ℃/h, the wet bulb temperature is adjusted to 37-38 ℃, and then the dry bulb temperature and the wet bulb temperature are stabilized and baked for 12-16 h until the top tobacco leaves are completely yellow; in the latter color fixing stage, after the initial color fixing stage is finished, the temperature of the dry bulb is increased to 53-54 ℃ at the heating rate of 0.5 ℃/h, the temperature of the wet bulb is increased to 38-39 ℃, and then the dry bulb and the wet bulb are stably baked for 20-24 h until the branches and the mesophyll of the tobacco leaves in the whole curing barn are dried;
e31, controlling the dry gluten period: after the E21 sub-step is finished, before the dry bulb temperature is 60 ℃, big fire is burnt, big moisture is discharged, then middle fire is gradually burnt, the moisture discharge amount of an air inlet is reduced, the dry bulb temperature is increased to 67-68 ℃ at the heating rate of 1 ℃/h, the wet bulb temperature is adjusted to 39-40 ℃, then the dry bulb temperature and the wet bulb temperature are stabilized for baking for 20-30 h until the main pulse of the tobacco leaves in the full baking room is dried, and finally the tobacco leaves are taken out of the furnace after being cooled along with the furnace.
And E, the baking of the K326 variety tobacco leaves in the step E comprises the steps of controlling the yellowing stage, controlling the fixation stage and controlling the dry stem stage, and the steps are as follows:
e12, yellowing stage control: the yellowing period comprises an initial yellowing period, a middle yellowing period and a later yellowing period, wherein in the initial yellowing period, the temperature of dry balls in a drying chamber of a curing barn is increased to 34-35 ℃ from room temperature and the temperature of wet balls is increased to 32-33 ℃ from room temperature at the heating rate of 1 ℃/1-2 h, and then the dry and wet balls are stably baked for 14-18 h; after the early yellowing stage is finished, the dry bulb temperature is increased to 38-40 ℃ at a heating rate of 1 ℃/1-2 h in the middle yellowing stage, the wet bulb temperature is adjusted to 36-37 ℃, and then the dry bulb temperature and the wet bulb temperature are stably baked for 20-22 h; in the later yellowing stage, after the middle yellowing stage is finished, the temperature of dry balls is increased to 42-43 ℃ at the temperature increase rate of 1 ℃/1-2 h, the temperature of wet balls is adjusted to be increased to 36-37 ℃, and then the dry and wet ball temperature is stabilized for baking for 14-18 h until the bottom table tobacco leaves are completely yellow;
e22, fixation period control: the ground color period comprises a color fixing initial stage and a color fixing later stage, wherein in the color fixing initial stage, after the E12 step is finished, the dry bulb temperature is increased to 45-47 ℃ at the heating rate of 1 ℃/1-2 h, the wet bulb temperature is adjusted to 36-38 ℃, and then the dry bulb temperature and the wet bulb temperature are stabilized for baking for 10-14 h until the top stage tobacco leaves are completely yellow; in the latter color fixing stage, after the initial color fixing stage is finished, the temperature of the dry bulb is increased to 55-57 ℃ at the temperature increasing rate of 1 ℃/1-2 h, the temperature of the wet bulb is increased to 38-40 ℃, and then the dry and wet bulb is stably baked for 24-30 h until the vein and the mesophyll of the tobacco leaves in the whole curing barn are dried;
e32, controlling the dry gluten period: after the E22 substep is finished, the dry bulb temperature is increased to 66-68 ℃ at the heating rate of 1 ℃/1-2 h, the wet bulb temperature is adjusted to 40-42 ℃, then the dry and wet bulb temperatures are stabilized for baking for 20-26 h until the main pulse of the tobacco leaves in the whole curing barn is dried, and finally the tobacco leaves are taken out of the curing barn after being cooled along with the curing barn.
And E, baking the KRK26 tobacco leaves in the step E, wherein the baking comprises the following steps of controlling the yellowing stage, controlling the fixation stage and controlling the dry stem stage:
e13, yellowing stage control: the yellowing period comprises an initial yellowing period, a middle yellowing period and a later yellowing period, wherein in the initial yellowing period, the temperature of dry balls in a drying chamber of a curing barn is increased to 34-35 ℃ from room temperature at the heating rate of 1 ℃/2h, the temperature of wet balls is increased to 33-34 ℃ from room temperature, and then the dry and wet ball temperatures are stabilized and cured for 10-14 h; after the early yellowing stage is finished, the dry bulb temperature is increased to 37-38 ℃ at the temperature rising rate of 1 ℃/2h in the middle yellowing stage, the wet bulb temperature is adjusted to 35-36 ℃, and then the dry bulb temperature and the wet bulb temperature are stably baked for 16-18 h; in the later yellowing stage, after the middle yellowing stage is finished, the temperature of dry balls is increased to 38-39 ℃ at the heating rate of 1 ℃/2h, the temperature of wet balls is adjusted to be increased to 35-37 ℃, and then the dry and wet balls are stably baked for 10-14 h until the bottom table tobacco leaves are completely yellow and the main veins are softened;
e23, fixation period control: the ground color period comprises a color fixing initial stage, a color fixing middle stage and a color fixing later stage, wherein in the color fixing initial stage, after the step E13 is finished, the dry bulb temperature is increased to 40-42 ℃ at the heating rate of 1 ℃/2h, the wet bulb temperature is adjusted to 36-37 ℃, and then the dry bulb temperature and the wet bulb temperature are stabilized and baked for 12-16 h until the branch veins are completely whitened and the leaves 1/3 are dried; after the color fixing initial stage is finished, the dry bulb temperature is increased to 46-48 ℃ at the heating rate of 1 ℃/h in the color fixing middle stage, the wet bulb temperature is adjusted to 36-38 ℃, and then the dry and wet bulb temperatures are stabilized for baking for 18-22 h until the main veins of the tobacco leaves in the whole baking room are completely whitened, and the leaves are dried above 2/3 and have an odor feeling; in the latter stage of color fixing, after the middle stage of color fixing is finished, the dry bulb temperature is increased to 54-56 ℃ and the wet bulb temperature is increased to 38-39 ℃ at the heating rate of 1 ℃/h, and then the dry and wet bulb temperatures are stabilized for baking for 10-14 h until the leaves of the tobacco leaves in the full curing barn are completely dried and the main vein stem of the high-temperature layer is 1/2 finished;
e33, controlling the dry gluten period: after the E23 substep is finished, the dry bulb temperature is increased to 65-67 ℃ at the heating rate of 1 ℃/h, the wet bulb temperature is adjusted to 39-40 ℃, then the dry and wet bulb temperatures are stabilized for baking for 26-30 h until the main pulse of the tobacco leaves in the full curing barn is dried, and finally the tobacco leaves are taken out of the furnace after being cooled along with the furnace.
The bulk curer is an airflow descending bulk curer.
Example 1 Effect of the baking Process of the present invention on the TSNAs content in Honghuadajinyuan tobacco leaves
S100: the method is characterized in that the safflower Honghuadajinyuan flue-cured tobacco fresh tobacco with proper maturity is harvested in Guanzhou province of Dalizhou, Yunnan province, and has the characteristics that the main vein is white and bright, the branch vein is dark and white, the leaves are green yellow and light yellow, most of fuzz falls off, and the tobacco leaves are rich in tobacco tar after being touched by hands.
S200: and (3) sorting and rod-weaving the collected fresh tobacco leaves according to the tobacco weaving amount of 90-110 leaves per rod.
S300: uniformly spraying and inoculating the bacterial liquid with the OD600 value of 1 on the surface of the tobacco leaves which are woven into the rods according to the amount of 3-5% of the weight of the tobacco leaves.
S400: and (3) loading the inoculated tobacco leaves with the braided rods into an airflow descending type bulk curing barn to more than 95% of the volume according to the tobacco loading amount of 540-580 rods/furnace and the tobacco loading density of 40-45 kg/m.
S500: and (4) baking the tobacco leaves after being charged into the furnace, and obtaining the tobacco leaves with low TSNAs after a yellowing period, a color fixing period and a stem drying period.
The bacterial liquid in S300 is prepared by adopting strains 05-101, J45 and J54 respectively. Wherein: t0 is clear water blank control solution without inoculating the above strain; preparing a T1 bacterial solution by adopting a 05-101 strain; preparing a T2 bacterial solution by using a J45 strain; preparing a T3 bacterial solution by using a J54 strain; the T4 bacterial liquid adopts three strains of 05-101, J45 and J54 according to the ratio of 1: 1: 1, mixing and preparing.
The yellowing period in S500 comprises a yellowing initial stage, a yellowing middle stage and a yellowing later stage, wherein the yellowing initial stage is to bake the dry-bulb temperature in a drying chamber of a curing barn from 26 ℃ to 33 ℃, the wet-bulb temperature from 24 ℃ to 32 ℃ and the stable temperature of the dry-bulb temperature and the stable temperature of the wet-bulb temperature for 18 hours at a heating rate of 1 ℃/h; then entering a yellowing middle stage, wherein the temperature of the dry bulb in the curing barn is increased to 39 ℃ at the temperature rising rate of 1 ℃/h, the temperature of the wet bulb is adjusted to be increased to 37 ℃, and the dry bulb and the wet bulb are cured for 26h at stable temperature; then, in the later yellowing stage, the temperature of the dry bulb in the drying chamber of the curing barn is increased to 42 ℃ at the heating rate of 1 ℃/h, the temperature of the wet bulb is adjusted to 38 ℃, the dry and wet bulb temperatures are stabilized and cured for 18h, and the tobacco leaves are cured to the bottom station to be completely yellow; the fixing period comprises a fixing early stage and a fixing later stage, wherein the drying ball temperature is increased to 47 ℃ at a heating rate of 0.5 ℃/h in the fixing early stage, the wet ball temperature is adjusted to 37 ℃, the drying and wet ball temperatures are stabilized, the tobacco leaves are baked for about 14h, and the tobacco leaves on the top stage are completely yellow; then, in the later stage of color fixing, the temperature of the dry bulb is increased to 53 ℃ at the heating rate of 0.5 ℃/h, the temperature of the wet bulb is adjusted to 38 ℃, the temperature of the dry bulb and the temperature of the wet bulb are stabilized, and the tobacco leaves are baked for 22h until the branches and the mesophyll of the tobacco leaves in the whole curing barn are dried; the dry stem drying period is that before the dry bulb temperature is 60 ℃, big fire is burnt, big moisture is discharged, then middle fire is gradually burnt, the moisture discharge amount of an air inlet is reduced, the dry bulb temperature is increased to 67 ℃ at the heating rate of 1 ℃/1h, the wet bulb temperature is adjusted to 39 ℃, then the dry bulb temperature and the wet bulb temperature are stabilized, the tobacco leaves are baked for 25h until the main vein of the tobacco leaves in the full curing barn is dried, and finally the tobacco leaves are taken out of the furnace after being cooled along with the furnace.
The above treatment was set as experimental group A, and experimental group B was set up on this basis.
Experimental group B: adjusting the wet bulb temperature to 38 ℃ in the yellowing period, and adjusting the baking time to 24 h; and in the later stage of the yellowing period, the temperature of the wet bulb is adjusted to be increased to 39 ℃, and the baking time is adjusted to be 20 h.
Experimental groups A, B were each treated with 5 treatments.
TABLE 1 TSNAs content comparison of cured tobacco after optimized technological process for variety of Hongda cured tobacco
Example 2 Effect of conventional baking Process on TSNAs content in Honghuadajinyuan tobacco leaves
A control experiment was set up on the basis of example 1: the wet bulb temperature in the yellowing period of the example 1 is adjusted to be 36 ℃, and the baking time is adjusted to be 22 h; adjusting the wet bulb temperature to 37 ℃ in the post-yellowing period, and adjusting the baking time to 14 h; other process parameters were unchanged.
TABLE 2 TSNAs content comparison of cured tobacco after conventional process for Honghuadajinyuan cured tobacco variety
As can be seen from Table 1, the TSNAs content of the cured tobacco is reduced by 21.29-23.97% under the condition of inoculating strains by using the safflower Hongjinyuan under the optimized process of the invention; as can be seen from Table 2, the TSNAs content of the flue-cured tobacco under the conventional process of the safflower Honghua Dajinyuan is reduced by 17.41-20.43% under the condition of inoculating strains, so that the inoculated strains can obviously reduce the TSNAs content of the flue-cured tobacco, and the optimized process of the invention has more obvious effect than the conventional process.
Example 3 Effect of the baking Process of the present invention on the Activity of the strains of Hongda tobacco leaves
According to examples 1 and 2, the viable cell count and NO of the bacterial species per leaf unit at the time of the completion of the early stage of yellowing in baking and the time of the late stage of yellowing2 -And (4) content.
TABLE 3 effective viable count and NO of tobacco leaves per unit area2 -Content comparison
As can be seen from Table 3, the process of the present method has significantly higher number of viable bacteria per unit area than the conventional process, and it eliminates NO which is a precursor substance of nitrosamine2 -The method has stronger capability, so the method proves that the process can improve the survival rate of the strains.
Example 4 Effect of the baking Process of the present invention on the TSNAs content in K326 tobacco leaves
S100: the method is characterized in that K326 flue-cured tobacco fresh tobacco leaves with proper maturity in Guanzhen town of Dali province of Yunnan province are collected, the fresh tobacco leaves with proper maturity have the characteristics that the main veins become white and bright, the branch veins turn green and white, the leaves are green yellow and light yellow, most of fuzz falls off, and the tobacco leaves are rich in tobacco tar after being touched by hands.
S200: and (3) sorting and rod-weaving the collected fresh tobacco leaves according to the tobacco weaving amount of 90-110 leaves per rod.
S300: uniformly spraying and inoculating the bacterial liquid with the OD600 value of 1 on the surface of the tobacco leaves which are woven into the rods according to the amount of 3-5% of the weight of the tobacco leaves.
S400: and (3) loading the inoculated tobacco leaves with the braided rods into an airflow descending type bulk curing barn to more than 95% of the volume according to the tobacco loading amount of 540-580 rods/furnace and the tobacco loading density of 40-45 kg/m.
S500: and (4) baking the tobacco leaves after being charged into the furnace, and obtaining the tobacco leaves with low TSNAs after a yellowing period, a color fixing period and a stem drying period.
The bacterial liquid in S300 is prepared by adopting strains 05-101, J45 and J54 respectively. Wherein: t0 is clear water blank control solution without inoculating the above strain; preparing a T1 bacterial solution by adopting a 05-101 strain; preparing a T2 bacterial solution by using a J45 strain; preparing a T3 bacterial solution by using a J54 strain; the T4 bacterial liquid adopts three strains of 05-101, J45 and J54 according to the ratio of 1: 1: 1, mixing and preparing.
The yellowing period in the S500 comprises an initial yellowing period, a middle yellowing period and a later yellowing period, wherein the initial yellowing period is to increase the temperature of a dry bulb from room temperature to 34 ℃ and adjust the temperature of a wet bulb from room temperature to 32 ℃ at a heating rate of 1 ℃/1-2 h, stabilize the temperature of the dry bulb and the temperature of the wet bulb, and bake for 16 h; then entering a yellowing middle stage, wherein the temperature of the dry bulb is raised to 39 ℃ at the heating rate of 1 ℃/1-2 h, the temperature of the wet bulb is adjusted to 36 ℃, and the dry bulb and the wet bulb are baked for 22h at a stable temperature; then, in the later yellowing stage, the temperature of the dry pellets is increased to 42 ℃ and the temperature of the wet pellets is adjusted to 36 ℃ at the heating rate of 1 ℃/1-2 h, the dry and wet pellet temperatures are stabilized, the tobacco leaves are baked for 16h, and the tobacco leaves are completely yellowed on the bottom platform; the fixing period comprises a fixing initial stage and a fixing later stage, wherein in the fixing initial stage, the dry bulb temperature is increased to 46 ℃ at a heating rate of 1 ℃/1-2 h, the wet bulb temperature is adjusted to 37 ℃, the dry bulb temperature and the wet bulb temperature are stabilized, and the top tobacco leaves are baked for 12h and completely turn yellow; then, in the later stage of color fixing, the temperature of the dry bulb is increased to 56 ℃ at the heating rate of 1 ℃/1-2 h, the temperature of the wet bulb is adjusted to 39 ℃, the temperature of the dry bulb and the temperature of the wet bulb are stabilized, and the tobacco leaves are baked for 28h until the branches and the mesophyll of the tobacco leaves in the whole curing barn are dried; in the stem drying period, the dry bulb temperature is increased to 67 ℃ at the heating rate of 1 ℃/1-2 h, the wet bulb temperature is adjusted to 41 ℃, the dry bulb temperature and the wet bulb temperature are stabilized, the tobacco leaves are baked for 24h until the main pulse of the tobacco leaves in the whole curing barn is dried, and finally the tobacco leaves are taken out of the curing barn after being cooled along with the curing barn.
The above treatment was set as experimental group A, and experimental group B was set up on this basis.
Experimental group B: adjusting the wet bulb temperature to 37 ℃ in the yellowing period, and adjusting the baking time to 20 h; and in the later stage of the yellowing period, the temperature of the wet bulb is adjusted to be increased to 37 ℃, and the baking time is adjusted to be 14 h.
Experimental groups A, B were each treated with 5 treatments.
TABLE 4 TSNAs content comparison of cured tobacco after optimized process for K326 cured tobacco variety
Example 5 Effect of conventional flue-curing Process on TSNAs content in K326 tobacco leaves
Control experiments were set up on the basis of example 4: the wet bulb temperature in the middle yellowing stage of example 4 is adjusted to 35 ℃, and the baking time is adjusted to 18 h; adjusting the wet bulb temperature to 35 ℃ in the post-yellowing period, and adjusting the baking time to 12 h; other process parameters were unchanged.
TABLE 5 TSNAs content comparison of cured tobacco after conventional process curing of K326 cured tobacco varieties
As can be seen from Table 4, the TSNAs content of the flue-cured tobacco with the optimized process of the invention is reduced by 21.37-23.90% under the condition of inoculating strains; as can be seen from Table 5, the TSNAs content of the flue-cured tobacco under the conventional K326 process is reduced by 16.88-20.32% under the condition of inoculating strains, so that the TSNAs content of the flue-cured tobacco can be obviously reduced by the inoculating strains, and the effect of the optimized process of the invention is more obvious than that of the conventional process.
Example 6 Effect of the baking Process of the present invention on the Activity of K326 tobacco Strain
According to examples 4 and 5, the viable cell count and NO of the bacterial species per leaf unit at the time of the completion of the early stage of yellowing in baking and the time of the late stage of yellowing2 -And (4) content.
TABLE 6 effective viable count and NO of tobacco leaves per unit area2 -Content comparison
As can be seen from Table 6, the process of the present method has significantly higher number of viable bacteria per unit area than the conventional process, and it eliminates NO which is a precursor of nitrosamine2 -The method has stronger capability, so the method proves that the process can improve the survival rate of the strains.
Example 7 Effect of the baking Process of the present invention on the TSNAs content of KRK26 tobacco leaves
S100: the KRK26 flue-cured tobacco fresh tobacco leaves with proper maturity of Guanzhen town under Dalizhou province of Yunnan province are collected, the fresh tobacco leaves with proper maturity have the characteristics that the main veins become white and bright, the branch veins turn green and white, the leaves are green yellow and light yellow, most of fuzz falls off, and the tobacco leaves are rich in tobacco tar after being touched by hands.
S200: and (3) sorting and rod-weaving the collected fresh tobacco leaves according to the tobacco weaving amount of 90-110 leaves per rod.
S300: uniformly spraying and inoculating the bacterial liquid with the OD600 value of 1 on the surface of the tobacco leaves which are woven into the rods according to the amount of 3-5% of the weight of the tobacco leaves.
S400: and (3) loading the inoculated tobacco leaves with the braided rods into an airflow descending type bulk curing barn to more than 95% of the volume according to the tobacco loading amount of 540-580 rods/furnace and the tobacco loading density of 40-45 kg/m.
S500: and (4) baking the tobacco leaves after being charged into the furnace, and obtaining the tobacco leaves with low TSNAs after a yellowing period, a color fixing period and a stem drying period.
The bacterial liquid in S300 is prepared by adopting strains 05-101, J45 and J54 respectively. Wherein: t0 is clear water blank control solution without inoculating the above strain; preparing a T1 bacterial solution by adopting a 05-101 strain; preparing a T2 bacterial solution by using a J45 strain; preparing a T3 bacterial solution by using a J54 strain; the T4 bacterial liquid adopts three strains of 05-101, J45 and J54 according to the ratio of 1: 1: 1, mixing and preparing.
The yellowing period in the S500 comprises a yellowing initial stage, a yellowing middle stage and a yellowing later stage, wherein the yellowing initial stage is to raise the dry bulb temperature from room temperature to 34 ℃ at a heating rate of 1 ℃/2h, adjust the wet bulb temperature from room temperature to 33 ℃, stabilize the dry bulb temperature and the wet bulb temperature and bake for 12 h; then entering a yellowing middle stage, wherein the temperature of the dry bulb is increased to 38 ℃ from the dry bulb temperature, the temperature of the wet bulb is adjusted to 35 ℃ at the heating rate of 1 ℃/2h in the yellowing middle stage, the dry bulb temperature and the wet bulb temperature are stabilized, and the baking is carried out for 18 h; then, in the later yellowing stage, the temperature of the dry bulb is increased to 39 ℃ at the heating rate of 1 ℃/2h, the temperature of the wet bulb is adjusted to 36 ℃, the temperature of the dry bulb and the temperature of the wet bulb are stabilized, the tobacco leaves are baked for 14h, the tobacco leaves on the bottom platform are completely yellowed, and the main veins are softened; the fixing period comprises a fixing initial stage, a fixing middle stage and a fixing later stage, wherein in the fixing initial stage, the dry bulb temperature is increased to 42 ℃ at a heating rate of 1 ℃/2h, the wet bulb temperature is adjusted to 36 ℃, the dry bulb temperature and the wet bulb temperature are stabilized, the mixture is baked for 14h until the branch veins are completely whitened, and the leaves are dried about 1/3; then, in the middle stage of color fixing, the temperature of the dry bulb is raised to 47 ℃ at the heating rate of 1 ℃/h, the temperature of the wet bulb is adjusted to 37 ℃, the temperature of the dry bulb and the temperature of the wet bulb are stabilized, the tobacco leaves are baked for 20h until the main veins of the tobacco leaves in the whole baking room are completely whitened, and the leaves are dried above 2/3 and have a smell; then, in the later stage of color fixing, the temperature of the dry bulb is increased to 55 ℃ at the heating rate of 1 ℃/1h, the temperature of the wet bulb is adjusted to 38 ℃, the temperature of the dry bulb and the temperature of the wet bulb are stabilized, the tobacco leaves in the whole curing barn are baked for 10-14 h, and the main vein of the high-temperature layer is 1/2; in the stem drying period, the dry bulb temperature is increased to 66 ℃ at the heating rate of 1 ℃/h, the wet bulb temperature is adjusted to 39 ℃, the dry bulb temperature and the wet bulb temperature are stabilized, the tobacco leaves are baked for 28h until the main pulse of the tobacco leaves in the whole curing barn is dried, and finally the tobacco leaves are taken out of the curing barn after being cooled along with the curing barn.
The above treatment was set as experimental group A, and experimental group B was set up on this basis.
Experimental group B: adjusting the wet bulb temperature to 36 ℃ in the yellowing period, and adjusting the baking time to 17 h; and in the later stage of the yellowing period, the temperature of the wet bulb is adjusted to be increased to 37 ℃, and the baking time is adjusted to be 10 h.
Experimental groups A, B were each treated with 5 treatments.
TABLE 7 TSNAs content comparison of cured tobacco after optimized process for KRK26 cured tobacco variety
Example 8 Effect of conventional flue-curing Process on TSNAs content in KRK26 tobacco leaves
Control experiments were set up on the basis of example 7: the wet bulb temperature in the yellowing period of example 7 was adjusted to 34 ℃ and the baking time was adjusted to 14 hours; adjusting the wet bulb temperature to 35 ℃ in the post-yellowing period, and adjusting the baking time to 12 h; other process parameters were unchanged.
TABLE 8 TSNAs content comparison of cured tobacco after conventional process KRK26
As can be seen from Table 7, the content of TSNAs of the flue-cured tobacco with KRK26 in the optimized process of the invention is reduced by 16.90-19.03% under the condition of inoculating strains; as can be seen from Table 8, the TSNAs content of the flue-cured tobacco under the conventional process of KRK26 is reduced by 20.69-22.78% under the condition of inoculating strains, so that the TSNAs content of the flue-cured tobacco can be obviously reduced by the inoculating strains, and the effect of the optimized process of the invention is more obvious than that of the conventional process.
Example 9 Effect of the baking Process of the present invention on the Activity of KRK26 tobacco Strain
According to examples 7 and 8, the viable cell count and NO of the bacterial species per leaf unit at the time of the completion of the early stage of yellowing in baking and the time of the late stage of yellowing2 -And (4) content.
TABLE 9 effective viable count and NO of tobacco leaves per unit area2 -Content comparison
As can be seen from Table 9, the number of viable bacteria per unit area is significantly higher and the purity is clearly lower in the process of the present method than in the conventional processNO as precursor for nitrosamine removal2 -The method has stronger capability, so the method proves that the process can improve the survival rate of the strains.
Claims (10)
1. A baking method for reducing TSNAs of different varieties of flue-cured tobaccos by using strains is characterized by comprising the steps of harvesting, rod weaving, inoculation, furnace charging and baking, and specifically comprises the following steps:
A. harvesting: collecting fresh flue-cured tobacco leaves with proper maturity;
B. rod weaving: classifying the collected fresh tobacco leaves and then weaving the tobacco leaves into rods;
C. inoculation: preparing bacterial liquid with an OD600 value of 1 by using at least one of the bacterial strains 05-101, J45 and J54, and then uniformly spraying the bacterial liquid to be inoculated on the surface of the tobacco leaves which are already woven into the rods;
D. charging: loading the inoculated tobacco leaves with the braided rods into a bulk curing barn;
E. baking: and (4) baking the tobacco leaves after being charged into the furnace, and obtaining the tobacco leaves with low TSNAs after a yellowing period, a color fixing period and a stem drying period.
2. The roasting method for reducing TSNAs of different varieties of flue-cured tobaccos by using strains according to claim 1, wherein the preservation number of the 05-101 strains is CCTCC NO: m2019665, the preservation number of the J45 strain is CCTCC NO: m2019666, the preservation number of the J54 strain is CCTCC NO: m2019667.
3. The curing method for reducing TSNAs of different varieties of flue-cured tobaccos by utilizing strains according to claim 2, wherein the amount of the strain solution with the OD600 value of 1 sprayed on the tobacco leaves in the step C is 3-5% of the weight of the tobacco leaves.
4. The curing method for reducing TSNAs of different varieties of flue-cured tobaccos by using strains according to claim 3, wherein the bacterial liquid is sprayed on the surface of the tobacco leaves in the step C and then the tobacco leaves are directly charged in a furnace and cured.
5. The curing method for reducing TSNAs of different varieties of flue-cured tobaccos by utilizing strains according to claim 2, wherein SPAD values of fresh tobaccos corresponding to suitable maturity of tobacco leaves at the lower part, the middle part and the upper part of a tobacco plant of the flue-cured tobaccos in the step A are respectively 8.6-11.4, 17.9-18.6 and 13.2-20.3, or the standard of the fresh tobaccos suitable for maturity is that angles of stems and leaves are 87-93 degrees, or the standard of the fresh tobaccos suitable for maturity is that main veins become white and bright, branch veins turn green and white, leaves are greenish yellow and yellowish, most of fuzz falls off, and the flue-cured tobaccos have rich tobacco oil after being touched by hands.
6. The curing method for reducing TSNAs of different varieties of flue-cured tobaccos by using strains according to any one of claims 1 to 5, wherein the varieties of the flue-cured tobaccos in the step A comprise Honghuadajinyuan, K326 and KRK 26.
7. The curing method for reducing TSNAs of different varieties of flue-cured tobaccos by utilizing strains according to claim 6, wherein the tobacco weaving amount of the tobacco leaves in the step B is 90-110 leaves/rod; and D, carrying out tobacco loading on the bulk curing barn by 540-580 rods/furnace, carrying out tobacco loading density by 40-45 kg/m and carrying out tobacco loading to more than 95% of the volume of the curing barn.
8. The roasting method for reducing TSNAs of different varieties of flue-cured tobaccos by utilizing strains according to claim 6, wherein the roasting of the Honghua Dajinyuan tobacco leaves in the step E comprises the following steps of yellowing stage control, color fixing stage control and tendon drying stage control, and the steps are as follows:
e11, yellowing stage control: the yellowing period comprises an initial yellowing period, a middle yellowing period and a later yellowing period, wherein in the initial yellowing period, the temperature of dry balls in a drying chamber of a curing barn is increased to 33-34 ℃ from room temperature at a heating rate of 1 ℃/h, the temperature of wet balls is increased to 32-33 ℃ from room temperature, and then the dry and wet balls are stably baked for 16-20 h; after the early yellowing stage is finished, the dry bulb temperature is increased to 38-39 ℃ at a heating rate of 1 ℃/h in the middle yellowing stage, the wet bulb temperature is adjusted to 37-38 ℃, and then the dry bulb temperature and the wet bulb temperature are stably baked for 24-28 h; in the later yellowing stage, after the middle yellowing stage is finished, the temperature of dry balls is increased to 42-43 ℃ at the heating rate of 1 ℃/h, the temperature of wet balls is adjusted to 38-39 ℃, and then the dry and wet balls are stably baked for 16-20 h until the bottom table tobacco leaves are completely yellow;
e21, fixation period control: the ground color period comprises a color fixing initial stage and a color fixing later stage, wherein in the color fixing initial stage, after the E11 step is finished, the dry bulb temperature is increased to 47-48 ℃ at the heating rate of 0.5 ℃/h, the wet bulb temperature is adjusted to 37-38 ℃, and then the dry bulb temperature and the wet bulb temperature are stabilized and baked for 12-16 h until the top tobacco leaves are completely yellow; in the latter color fixing stage, after the initial color fixing stage is finished, the temperature of the dry bulb is increased to 53-54 ℃ at the heating rate of 0.5 ℃/h, the temperature of the wet bulb is increased to 38-39 ℃, and then the dry bulb and the wet bulb are stably baked for 20-24 h until the branches and the mesophyll of the tobacco leaves in the whole curing barn are dried;
e31, controlling the dry gluten period: after the E21 sub-step is finished, before the dry bulb temperature is 60 ℃, big fire is burnt, big moisture is discharged, then middle fire is gradually burnt, the moisture discharge amount of an air inlet is reduced, the dry bulb temperature is increased to 67-68 ℃ at the heating rate of 1 ℃/h, the wet bulb temperature is adjusted to 39-40 ℃, then the dry bulb temperature and the wet bulb temperature are stabilized for baking for 20-30 h until the main pulse of the tobacco leaves in the full baking room is dried, and finally the tobacco leaves are taken out of the furnace after being cooled along with the furnace.
9. The roasting method for reducing TSNAs of different varieties of flue-cured tobaccos by utilizing strains according to claim 6, wherein the roasting of the K326 variety of tobacco leaves in the E step comprises the following steps of yellowing stage control, color fixing stage control and tendon drying stage control, and the steps are as follows:
e12, yellowing stage control: the yellowing period comprises an initial yellowing period, a middle yellowing period and a later yellowing period, wherein in the initial yellowing period, the temperature of dry balls in a drying chamber of a curing barn is increased to 34-35 ℃ from room temperature and the temperature of wet balls is increased to 32-33 ℃ from room temperature at the heating rate of 1 ℃/1-2 h, and then the dry and wet balls are stably baked for 14-18 h; after the early yellowing stage is finished, the dry bulb temperature is increased to 38-40 ℃ at a heating rate of 1 ℃/1-2 h in the middle yellowing stage, the wet bulb temperature is adjusted to 36-37 ℃, and then the dry bulb temperature and the wet bulb temperature are stably baked for 20-22 h; in the later yellowing stage, after the middle yellowing stage is finished, the temperature of dry balls is increased to 42-43 ℃ at the temperature increase rate of 1 ℃/1-2 h, the temperature of wet balls is adjusted to be increased to 36-37 ℃, and then the dry and wet ball temperature is stabilized for baking for 14-18 h until the bottom table tobacco leaves are completely yellow;
e22, fixation period control: the ground color period comprises a color fixing initial stage and a color fixing later stage, wherein in the color fixing initial stage, after the E12 step is finished, the dry bulb temperature is increased to 45-47 ℃ at the heating rate of 1 ℃/1-2 h, the wet bulb temperature is adjusted to 36-38 ℃, and then the dry bulb temperature and the wet bulb temperature are stabilized for baking for 10-14 h until the top stage tobacco leaves are completely yellow; in the latter color fixing stage, after the initial color fixing stage is finished, the temperature of the dry bulb is increased to 55-57 ℃ at the temperature increasing rate of 1 ℃/1-2 h, the temperature of the wet bulb is increased to 38-40 ℃, and then the dry and wet bulb is stably baked for 24-30 h until the vein and the mesophyll of the tobacco leaves in the whole curing barn are dried;
e32, controlling the dry gluten period: after the E22 substep is finished, the dry bulb temperature is increased to 66-68 ℃ at the heating rate of 1 ℃/1-2 h, the wet bulb temperature is adjusted to 40-42 ℃, then the dry and wet bulb temperatures are stabilized for baking for 20-26 h until the main pulse of the tobacco leaves in the whole curing barn is dried, and finally the tobacco leaves are taken out of the curing barn after being cooled along with the curing barn.
10. The curing method for reducing TSNAs of different varieties of flue-cured tobaccos by utilizing strains according to claim 6, wherein the curing of the tobacco leaves of the KRK26 variety in the step E comprises the following steps of yellowing stage control, color fixing stage control and tendon drying stage control:
e13, yellowing stage control: the yellowing period comprises an initial yellowing period, a middle yellowing period and a later yellowing period, wherein in the initial yellowing period, the temperature of dry balls in a drying chamber of a curing barn is increased to 34-35 ℃ from room temperature at the heating rate of 1 ℃/2h, the temperature of wet balls is increased to 33-34 ℃ from room temperature, and then the dry and wet ball temperatures are stabilized and cured for 10-14 h; after the early yellowing stage is finished, the dry bulb temperature is increased to 37-38 ℃ at the temperature rising rate of 1 ℃/2h in the middle yellowing stage, the wet bulb temperature is adjusted to 35-36 ℃, and then the dry bulb temperature and the wet bulb temperature are stably baked for 16-18 h; in the later yellowing stage, after the middle yellowing stage is finished, the temperature of dry balls is increased to 38-39 ℃ at the heating rate of 1 ℃/2h, the temperature of wet balls is adjusted to be increased to 35-37 ℃, and then the dry and wet balls are stably baked for 10-14 h until the bottom table tobacco leaves are completely yellow and the main veins are softened;
e23, fixation period control: the ground color period comprises a color fixing initial stage, a color fixing middle stage and a color fixing later stage, wherein in the color fixing initial stage, after the step E13 is finished, the dry bulb temperature is increased to 40-42 ℃ at the heating rate of 1 ℃/2h, the wet bulb temperature is adjusted to 36-37 ℃, and then the dry bulb temperature and the wet bulb temperature are stabilized and baked for 12-16 h until the branch veins are completely whitened and the leaves 1/3 are dried; after the color fixing initial stage is finished, the dry bulb temperature is increased to 46-48 ℃ at the heating rate of 1 ℃/h in the color fixing middle stage, the wet bulb temperature is adjusted to 36-38 ℃, and then the dry and wet bulb temperatures are stabilized for baking for 18-22 h until the main veins of the tobacco leaves in the whole baking room are completely whitened, and the leaves are dried above 2/3 and have an odor feeling; in the latter stage of color fixing, after the middle stage of color fixing is finished, the dry bulb temperature is increased to 54-56 ℃ and the wet bulb temperature is increased to 38-39 ℃ at the heating rate of 1 ℃/h, and then the dry and wet bulb temperatures are stabilized for baking for 10-14 h until the leaves of the tobacco leaves in the full curing barn are completely dried and the main vein stem of the high-temperature layer is 1/2 finished;
e33, controlling the dry gluten period: after the E23 substep is finished, the dry bulb temperature is increased to 65-67 ℃ at the heating rate of 1 ℃/h, the wet bulb temperature is adjusted to 39-40 ℃, then the dry and wet bulb temperatures are stabilized for baking for 26-30 h until the main pulse of the tobacco leaves in the full curing barn is dried, and finally the tobacco leaves are taken out of the furnace after being cooled along with the furnace.
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