Compound enzyme preparation for improving combustibility of Nanping cigars and application of compound enzyme preparation

Abstract

The invention belongs to the field of biological enzyme preparations, and particularly relates to a compound enzyme preparation for improving combustibility of Nanping cigars and application of the compound enzyme preparation. According to the method, the Nanping cigar tobacco leaves with improved combustion performance are prepared by harvesting the Nanping cigars, airing the Nanping cigars, screening and compounding enzyme preparations, optimizing an enzymolysis mode and controlling enzymolysis parameters only by adopting a compound enzyme preparation without non-enzyme preparations. Pectate lyase and pectin esterase in pectinase, papain in protease, beta-amylase saccharifying enzyme and isoamylase in amylase and cellulase are screened out and combined, and further optimized that pectate lyase, pectate esterase and cellulase are subjected to mixed enzymolysis, enzymolysis is carried out, enzymolysis is carried out, and enzymolysis is carried out. The papain, beta-amylase and isoamylase solutions are mixed for enzymolysis, and the temperature and time of two times of enzymolysis are controlled, so that the combustibility of the Nanping cigar tobacco leaves is remarkably improved, and meanwhile, the smoking evaluation result of the Nanping cigar tobacco leaves is good.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biological enzyme preparation, and particularly relates to a compound enzyme preparation for improving the combustibility of Nanping cigar and application thereof. BACKGROUND

[0002] Cigar, also known as cigar tobacco, is a kind of cigarette, which is made of dried and fermented tobacco. Cigar is an early product in the production of tobacco products, but the production of cigar in China is relatively late.

[0003] Cigar combustibility refers to the performance of cigar tobacco or finished cigar when it is ignited and burns without flame. Cigar combustibility has a significant impact on smoke quality. The indicators for measuring the combustibility of cigar include two aspects of smoldering and ash color, wherein the smoldering indicators include four aspects of smoldering time, smoldering rate, combustion completeness and combustion uniformity. The linear burning rate or mass burning rate of cigar when it is smoldering, and the smoldering time are usually used to describe the combustibility of Nanping cigar.

[0004] The combustibility of cigar is the result of the interaction of various chemical components in the cigar at high temperature. Current research shows that the main chemical components affecting the combustibility of cigar include total sugar and reducing sugar content, plant alkaloid, total nitrogen content, volatile acid, starch, cellulose, pectin content, and potassium-chlorine ratio. Part of the existing technology records that the total sugar content, reducing sugar content and the difference between the two sugar contents are significantly positively correlated with the combustibility, while the protein, plant alkaloid and total nitrogen content are extremely significantly negatively correlated with the combustibility. When the sugar-alkali ratio and the nitrogen-alkali ratio are high, the combustibility is better. Part of the existing technology records that the correlation between the two sugar ratio and the combustibility is not significant. When the total potassium content in the cigar is increased, the cigar has lighter miscellaneous gas and improved combustibility, and the ash color is whiter. When the chlorine content in the cigar is too high, the combustibility is reduced and the ash color is poor. When the volatile acid content in the cigar is increased, the score of the combustibility of the cigar is significantly improved, and when the volatile alkali content is increased, the score of the combustibility is significantly reduced. Under the same combustion conditions, starch and cellulose have higher flammability than pectin, and the combustibility of cellulose is better than that of starch, while the combustibility of pectin is relatively poor. When the starch content in the cigar is increased, the combustibility of the cigar will be significantly reduced. Starch has good coke formation property and releases more non-combustible gas products, thereby reducing the combustibility of Nanping cigar. Cellulose is beneficial to the combustibility of Nanping cigar, but too high cellulose content will produce a choking odor and make the cigar organization rough and fragile.

[0005] Harvesting and curing are important links in the production process of Nanping cigar tobacco. Suitable harvesting maturity and reasonable temperature and humidity settings can coordinate and promote the internal chemical composition and material transformation of Nanping cigar tobacco, thereby improving the combustibility.

[0006] According to the search, there are enzyme preparations for treating tobacco, changing the content of total sugar and reducing sugar in tobacco, plant alkaline, total nitrogen content, volatile acid, starch, cellulose, pectin content and or potassium chloride ratio, etc., so as to improve the combustion of tobacco, such as non-patent document 1 ("Effect of exogenous enzyme preparation on aroma quality and combustion characteristics of flue-cured tobacco", Fan Wenju, Southwestern Agricultural Sciences, Vol. 31, No. 5, pp. 948-957, 2018) discloses that YN99 tobacco leaves are used as test materials in Qinling tobacco area, and 7 kinds of exogenous enzymes such as neutral protease, papain, flavor protease, cellulase, pectinase, saccharifying enzyme and alpha-amylase are set. However, this document only uses single enzyme preparation treatment, and does not study the combination of enzyme preparation and the mode of enzymolysis. The object of treatment is also not Nanping cigar; patent documents also exist for the improvement of enzyme preparation on tobacco or Nanping cigar tobacco leaf combustion, such as CN104939293A discloses a method for fermenting Nanping cigar tobacco leaf with pectinase, which only uses pectinase. Similarly, the single enzyme preparation has poor effect, and the influence of different models of pectinase on Nanping cigar tobacco leaf is not studied; CN103120362A discloses a method for improving the combustion of Nanping cigar tobacco leaf, which only uses single saccharifying enzyme, and the effect is not good; CN114403496A discloses a synergist for improving the combustion and ash color of fermented Nanping cigar tobacco leaf and a method for using the same, which only uses protease, and also adds reducing sugar and non-enzyme preparation means such as combustion enhancer. At present, there is no literature on the influence of Nanping cigar harvesting maturity and curing process on cigar tobacco leaf combustion.

[0007] The prior art does not study the influence of Nanping cigar harvesting maturity and curing process on cigar tobacco leaf combustion, and there is no systematic study on the complex enzyme preparation suitable for improving the combustion of Nanping cigar tobacco leaf. There is no complex enzyme preparation only by the combination of enzyme preparation, the optimization of enzymolysis mode and the control of enzymolysis parameters to improve the combustion of Nanping cigar tobacco leaf. SUMMARY

[0008] In one aspect, the present application provides a method for improving the combustion of Nanping cigar tobacco leaf by harvesting maturity and curing process. The method comprises the following steps: preparing Nanping cigar tobacco leaf by adopting M1LTHH process, wherein the harvesting maturity of the Nanping cigar tobacco leaf is green, and the curing process comprises the following steps: in the wilting stage, the humidity is 95-98%RH, and the temperature is 22-25℃; in the yellowing stage, the humidity is 90-95%RH, and the temperature is 25-28℃; in the browning stage, the humidity is 85-90%RH, and the temperature is 28-31℃; in the color fixing stage, the humidity is 70-80%RH, and the temperature is 31-34℃; in the dry muscle stage, the humidity is 60-70%RH, and the temperature is 31-34℃.

[0009] The second aspect of the present application provides a complex enzyme preparation for improving the combustibility of Nanping cigar tobacco leaves, wherein the complex enzyme preparation comprises pectin lyase, pectin esterase, papain, beta-amylase, isoamylase and cellulase solution; the enzyme preparation solution is prepared by adding pectin lyase, pectin esterase, papain, beta-amylase, isoamylase and cellulase into pure water respectively to form enzyme preparation solutions with a mass concentration of 2%; and the Nanping cigar tobacco leaves are prepared by using the M1LTHH process, i.e., the harvested Nanping cigar tobacco leaves are green in maturity, and the curing process is as follows: wilting period, humidity 95-98% RH, temperature 22-25℃; yellowing period, humidity 90-95% RH, temperature 25-28℃; browning period, humidity 85-90% RH, temperature 28-31℃; color fixing period, humidity 70-80% RH, temperature 31-34℃; and dry period, humidity 60-70% RH, temperature 31-34℃.

[0010] The third aspect of the present application provides an application of the complex enzyme preparation for improving the combustibility of Nanping cigar tobacco leaves in the preparation of Nanping cigar tobacco leaves.

[0011] The fourth aspect of the present application provides a method for improving the combustibility of Nanping cigar tobacco leaves by using a complex enzyme preparation for controllable enzymolysis, which comprises the following steps: (1) adding pectin lyase, pectin esterase, papain, beta-amylase, isoamylase and cellulase into pure water respectively to form enzyme preparation solutions with a mass concentration of 2%; (2) mixing the pectin lyase, pectin esterase and cellulase solutions, mixing the papain, beta-amylase and isoamylase solutions, placing the Nanping cigar tobacco leaves in the mixed enzyme preparation solution of pectin lyase, pectin esterase and cellulase, and performing primary enzymolysis at 35-45℃ for 25-35 minutes, taking out the Nanping cigar tobacco leaves, inactivating the enzymes, placing the Nanping cigar tobacco leaves in the mixed enzyme preparation solution of papain, beta-amylase and isoamylase, and performing secondary enzymolysis at 25-35℃ for 20-30 minutes, taking out the Nanping cigar tobacco leaves and drying, and controlling the water content of the Nanping cigar tobacco leaves to be 12%.

[0012] Further, the temperature for the primary enzymolysis in step (2) is 40-45℃, preferably 45℃, and more preferably 40℃.

[0013] Further, the time for the primary enzymolysis in step (2) is 30-35 minutes, preferably 30 minutes.

[0014] Further, the temperature for the secondary enzymolysis in step (2) is 30-35℃, preferably 30℃.

[0015] Further, the time for the secondary enzymolysis in step (2) is 25-30 minutes, preferably 25 minutes. The fourth aspect of the present application provides a method for improving the combustibility of Nanping cigar tobacco leaves by using a complex enzyme preparation to control enzymolysis, comprising the following steps: (1) Take pectin lyase, pectin esterase, papain, beta-amylase, isoamylase, and cellulase, respectively, add them to pure water, and prepare enzyme preparation solutions with a mass concentration of 2% respectively; (2) Mix the pectin lyase, pectin esterase, and cellulase solutions, mix the papain, beta-amylase, and isoamylase solutions, place the Nanping cigar tobacco leaves in the mixed enzyme preparation solution of pectin lyase, pectin esterase, and cellulase, and perform enzymolysis at 40°C for 30 minutes, then take out the leaves, kill the enzymes, and place them in the mixed enzyme preparation solution of papain, beta-amylase, and isoamylase, and perform enzymolysis at 30°C for 25 minutes, then take out the leaves, dry them, and control the moisture content of the Nanping cigar tobacco leaves to be 12%.

[0016] The Nanping cigar tobacco leaves with improved combustibility prepared by any of the above-mentioned methods.

[0017] Compared with the prior art, the present specification achieves at least the following beneficial effects: (1) The present specification systematically studies the effects of harvesting maturity and curing process on Nanping cigar tobacco leaves, and the results show that the Nanping cigar tobacco leaves prepared by the M1LTHH process (harvesting maturity is green, and the curing process is: wilting period humidity 95-98%RH, temperature 22-25°C, yellowing period humidity 90-95%RH, temperature 25-28°C, browning period humidity 85-90%RH, temperature 28-31°C, color fixing period humidity 70-80%RH, temperature 31-34°C, dry tendon period humidity 60-70%RH, temperature 31-34°C) or the M2LTHH process can improve the combustibility thereof.

[0018] (2) The present specification selects pectin lyase and pectin esterase from pectinase, papain from protease, beta-amylase saccharifying enzyme and isoamylase from amylase, and cellulase from cellulase, which have an impact on the combustibility of Nanping cigar tobacco leaves, combines them, and significantly improves the combustibility of Nanping cigar tobacco leaves by only optimizing the enzyme hydrolysis method and controlling the enzyme hydrolysis conditions without using non-enzyme preparation means.

[0019] (3) The present specification studies the effects of different enzyme hydrolysis methods on the combustibility of Nanping cigar tobacco leaves, and selects the method of first mixing pectin lyase, pectin esterase, and cellulase for enzyme hydrolysis, and then mixing papain, beta-amylase, and isoamylase solution for enzyme hydrolysis, which can significantly improve the combustibility of Nanping cigar tobacco leaves. The possible reason is that pectin lyase, pectin esterase, and cellulase are all wall-breaking enzymes, which can damage the cell wall of plants and make the internal structure loose, so that papain, beta-amylase, and isoamylase can more easily combine with their respective substrates, thereby improving the efficiency of enzyme hydrolysis.

[0020] (4) The present application significantly improves the combustibility of Nanping cigar tobacco leaves by controlling the first enzymolysis temperature and time, and the second enzymolysis temperature and time. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 Gray comparison of cigar tobacco leaves of different harvesting maturities.

[0022] Figure 2 Gray comparison of cigar tobacco leaves of different curing temperatures.

[0023] Figure 3 Gray comparison of cigar tobacco leaves of different curing humidities.

[0024] Figure 4 Gray comparison of cigar tobacco leaves of different harvesting maturities and curing treatments.

[0025] Figure 5 Example 4: Effect of first enzymolysis temperature on combustibility of Nanping cigar tobacco leaves.

[0026] Figure 6 Example 4: Effect of first enzymolysis time on combustibility of Nanping cigar tobacco leaves.

[0027] Figure 7 Example 4: Effect of second enzymolysis temperature on combustibility of Nanping cigar tobacco leaves.

[0028] Figure 8 Example 4: Effect of second enzymolysis time on combustibility of Nanping cigar tobacco leaves. DETAILED DESCRIPTION

[0029] The specific embodiments of the present application are described below in conjunction with the accompanying drawings. In the implementation examples, the experimental methods used are conventional methods unless otherwise specified; and the materials, reagents, etc. used are commercially available unless otherwise specified.

[0030] Example 1: Effect of harvesting and curing process on combustibility of Nanping cigar tobacco leaves In combination with the environmental conditions such as precipitation, temperature, and humidity in Nanping tobacco-growing areas, cigar tobacco leaves of different maturities were collected, and different temperature and humidity technical parameters during curing were set. The interaction of different harvesting maturities and temperature and humidity conditions during different curing stages on the internal chemical components and their transformation of cigar tobacco leaves was studied, and the effects on combustibility, ash content, and sensory evaluation quality were determined to determine the most suitable harvesting maturity of cigar tobacco leaves and the matching key temperature and humidity technical parameters during curing.

[0031] Test site The experiment was conducted in the tea-fruit field of the San-yuan village of the Yan-shan town of the Shao-wu city of the Nan-ping city. The uniform growth of the cigar tobacco field is required for the picking of the cigar tobacco leaves with different maturity. The interactive contrast experiment of the picking maturity and the curing process was carried out in the cigar tobacco curing barn of the tobacco experimental station.

[0032] Test design The double-factor experiment design of the picking maturity and the curing process was adopted. The picking maturity was set as one part and three maturity treatments. The part was the middle tobacco leaves (8-12 leaf positions). The maturity was judged by the color, which was green (M1), green-yellow (M2) and yellow-green (M3) respectively. The curing process was set as four treatments, i.e. low temperature and high humidity (LTHH), medium temperature and high humidity (MTHH), low temperature and medium humidity (LTMH) and medium temperature and medium humidity (MTMH). There were 12 treatments in total. The picking maturity of the cigar tobacco leaves was determined by the color appearance. The curing process was adjusted by the placing position and the curing density of the tobacco leaves in the curing barn.

[0033] Table 1. The experiment treatment table of the picking maturity and the curing temperature and humidity Table 2. The setting reference table of the curing temperature and humidity Chemical component index determination The chemical components of the tobacco leaves were determined, which mainly included the regular chemical components (nicotine, total nitrogen, total sugar, reducing sugar, potassium, chlorine), the microelements (calcium, magnesium), the carbohydrates (starch, lignin, cellulose) and the secondary alkaloids.

[0034] Effects of different picking maturity on the chemical components of the cigar tobacco leaves From the Tables 3 and 4, the different picking maturity had different effects on the chemical components of the cigar tobacco leaves, in which the individual indexes reached the extremely significant difference between the treatments. With the increase of the picking maturity, the contents of the total nitrogen, total plant alkaloids, calcium and the potassium-chlorine ratio showed the gradually increasing trend, and the difference between the treatments was not significant. The contents of the total sugar, reducing sugar, potassium and chlorine showed the gradually decreasing trend, and the difference between the treatments was not significant. The contents of the nicotine and the pseudotuberfractin showed the trend of first decreasing and then increasing, and there was the extremely significant difference between the treatments. The contents of the magnesium, starch, cellulose, lignin, nitrogen-alkaloid ratio and sugar-alkaloid ratio did not show the obvious trend, and the difference between the treatments was not significant.

[0035] Table 3. The regular chemical components of different picking maturity Table 4. The other chemical components of different picking maturity Effects of different curing process on the chemical components of the cigar tobacco leaves From table 5 and 6, different curing process has different degree of influence on the chemical composition of cigar tobacco leaf, in which individual index reaches extremely significant difference between treatments. With the increase of curing temperature and humidity, the content of total plant alkaloid and lignin presents a small upward trend, in which total plant alkaloid exists extremely significant difference between treatments; the content of total nitrogen, starch, cellulose and nitrogen alkaline ratio presents a small downward trend, in which total plant alkaloid and nitrogen alkaline ratio exists extremely significant difference between treatments; the content of total sugar, reducing sugar, potassium, chlorine, calcium, magnesium, secondary alkaloid and sugar alkaline ratio, potassium chlorine ratio does not present obvious trend, and the difference between treatments is not significant.

[0036] Table 5 General chemical composition of different curing process Table 6 Other chemical composition of different curing process Effect of harvest maturity and curing process on the chemical composition of cigar tobacco leaf From table 7 and 8, the interaction of different harvest maturity and different curing process has different degree of influence on the chemical composition of cigar tobacco leaf, in which total nitrogen, total plant alkaloid, potassium, calcium, starch, lignin, nicotine, anabasine, false ephedra alkaline and nitrogen alkaline ratio, potassium chlorine ratio reaches extremely significant difference between treatments, total sugar, reducing sugar, chlorine, magnesium, cellulose, nicotine and sugar alkaline ratio reaches significant difference between treatments.

[0037] Table 7 General chemical composition of harvest maturity and curing process interaction The total nitrogen content of M3MTHH treatment was the highest, M2MTHH treatment was the second, and M2LTHH treatment was the lowest; the total plant base content of M1MTHH treatment was the highest, M2LTMH treatment and M2LTHH treatment were the second, and M2MTMH treatment was the lowest; the total sugar content of M2MTMH treatment was the highest, M1MTHH treatment and M2MTHH treatment were the second, and M3LTHH treatment was the lowest; the reducing sugar content of M3MTMH treatment was the highest, M1LTMH treatment was the second, and M1LTHH treatment was the lowest; the potassium content of M1MTHH treatment was the highest, M1MTMH treatment was the second, and M2MTMH treatment was the lowest; the chlorine content of M2LTHH treatment was the highest, M2MTHH treatment and M2MTMH treatment were the second, and M2LTMH treatment was the lowest; the nitrogen base ratio of M3MTHH treatment was the largest, M3LTMH treatment and M3MTMH treatment were the second, and M2LTHH treatment was the smallest; the sugar base ratio of M3MTMH treatment was the largest, M1LTMH treatment was the second, and M2LTHH treatment was the smallest; the potassium chlorine ratio of M1MTHH treatment was the largest, M3MTHH treatment was the second, and M2MTMH treatment was the smallest.

[0038] Table 8 Other chemical components of the interaction of harvesting maturity and curing process The calcium content of M3LTMH treatment was the highest, M1LTMH treatment was the second, and M2MTMH treatment was the lowest; the magnesium content of M2MTMH treatment was the highest, M3LTMH treatment was the second, and M1LTMH treatment was the lowest; the starch content of M2LTMH treatment was the highest, M3LTHH treatment and M2LTHH treatment were the second, and M1MTMH treatment was the lowest; the cellulose content of M1LTMH treatment was the highest, M3MTHH treatment was the second, and M3LTHH treatment was the lowest; the lignin content of M2MTMH treatment was the highest, M2MTHH treatment and M3LTHH treatment were the second, and M2MTMH treatment was the lowest; the nicotine content of M2LTHH treatment was the highest, M3LTMH treatment and M1MTHH treatment were the second, and M2MTMH treatment was the lowest; the nornicotine content of M3MTHH treatment was the highest, M2MTMH treatment and M1MTHH were the second, and M3LTMH treatment was the lowest; the anabasine content of M2LTHH treatment was the highest, M2MTHH treatment and M3LTHH treatment were the second, and M2LTHH treatment was the lowest.

[0039] Effects of different harvesting maturity on the combustibility and ash of cigar tobacco leaves Table 9 shows that different harvest maturity levels have varying degrees of influence on the combustibility and ash content of cigar tobacco leaves, with significant differences observed among treatments for each index. Brightness ( The ash content and whiteness showed a trend of first decreasing and then increasing with the increase of harvest maturity, with M1 having the highest ash brightness and the color closest to white-gray; the burning rate showed a trend of gradually decreasing with the increase of harvest maturity, with M1 having the highest burning rate.

[0040] Table 9 Combustibility and Ash Content at Different Harvest Maturities The Influence of Different Drying Processes on the Combustibility and Ash Content of Cigar Tobacco Leaves Table 10 shows that different drying processes have varying degrees of impact on the combustibility and ash content of cigar tobacco leaves, with significant differences observed among the treatments. Brightness ( The ash content and whiteness gradually decrease with increasing temperature, but gradually increase with increasing humidity. LTHH has the highest ash brightness and the color closest to white-gray. The burning rate gradually decreases with increasing temperature and humidity, while LTMH has the highest burning rate.

[0041] Table 10 Combustibility and ash content of different drying processes The Influence of Harvest Maturity and Drying Process on the Combustibility and Ash Content of Cigar Tobacco Leaves As shown in Table 11, under the interaction of different harvest maturity levels and different drying processes, the combustibility and ash content of cigar tobacco leaves changed to varying degrees. Brightness, whiteness, and burning rate showed highly significant differences among the treatments. The M1LTHH treatment had the highest ash brightness, followed by the M2LTHH and M3MTMH treatments, while the M3LTMH treatment had the lowest ash brightness. The M1MTHH treatment had the highest ash whiteness, followed by the M1LTHH and M2LTMH treatments, while the M3MTMH treatment had the lowest ash whiteness. The M3MTMH treatment had the highest burning rate, followed by the M1LTHH and M3LTHH treatments, while the M3MTMH treatment had the lowest burning rate.

[0042] Table 11 Combustibility and Ash Content of Harvest Maturity and Drying Process The Influence of Harvest Maturity and Drying Process on the Sensory Quality of Cigar Tobacco Leaves From Table 12, it can be seen that different harvesting maturities and curing processes have different effects on the aroma characteristics, smoke characteristics, aftertaste characteristics and combustion characteristics of cigar leaves. M2LTMH and M3MTMH perform best in terms of aroma characteristics, followed by M1LTMH, M1MTHH, etc.; M1LTMH and M2MTHH perform best in terms of smoke characteristics, followed by M1MTHH, M2LTMH, etc.; M1MTHH, M2MTMH and M2MTHH perform best in terms of aftertaste characteristics, followed by M1LTMH, M1LTHH, etc.; M1LTHH, M1MTMH and M2LTHH perform best in terms of combustion characteristics, followed by M3LTMH, M3LTHH, etc.; the overall sensory quality score of M1LTHH and M2LTHH is the highest, followed by M1MTHH, M3LTHH and M2MTHH, etc. The overall description of the sensory quality of M1LTHH and M2LTHH is that the aroma amount is sufficient, the richness is high, the maturity is good, the smoke is delicate and soft, the irritation is small, the aftertaste is good, the taste is clean and the sweet feeling is strong, and the combustion is good, the ash is grayish white, and the ash condensation degree is high.

[0043] Table 12 Sensory quality of cigar leaves with different harvesting maturities and curing processes According to the feedback of cigar sensory evaluation experts, the treatment with harvesting maturity M1 and low-temperature high-humidity curing process has no obvious alkaline odor, followed by the treatment with harvesting maturity M2 and low-temperature high-humidity curing process, and the other treatments have slight or slightly alkaline odor.

[0044] In summary, the green (M1) harvesting maturity and the low-temperature high-humidity (LTHH) curing process, i.e. the M1LTHH process (harvesting maturity green, curing process: wilting period humidity 95-98%RH, temperature 22-25℃, yellowing period humidity 90-95%RH, temperature 25-28℃, browning period humidity 85-90%RH, temperature 28-31℃, color fixing period humidity 70-80%RH, temperature 31-34℃, dry period humidity 60-70%RH, temperature 31-34℃) or the M2LTHH process are selected to prepare Nanping cigar leaves.

[0045] Example 2 Effect of different enzyme preparations on the combustion of Nanping cigar leaves According to the prior art, the main chemical components affecting the combustion of Nanping cigar include total sugar and reducing sugar content, plant alkaloid, total nitrogen content, volatile acid, starch, cellulose, pectin content, potassium-chlorine ratio, etc., and pectinase, protease, amylase, cellulase and hemicellulase all affect the above components. Therefore, the above enzyme preparations are selected to study their effects on the combustion of Nanping cigar leaves (prepared by the M1LTHH process), and the specific steps are as follows: (1) Take different enzyme preparations: pectinase (pectin lyase, polygalacturonase, pectin esterase), protease (papain, alkaline protease, neutral protease, flavor protease), amylase (a-amylase, b-amylase, saccharifying enzyme, isoamylase), cellulase, hemicellulase, respectively added in pure water, prepared into enzyme preparation solution with mass concentration of 2%; (2) Put Nanping cigar tobacco leaves in enzyme preparation solution respectively, and carry out enzymolysis at 35℃ for 30 minutes, take out and dry, and control the moisture content of Nanping cigar tobacco leaves to be 12%. The specific determination results are shown in Table 13.

[0046] Table 13 According to Table 13, under the same enzyme preparation addition amount, pectin lyase and pectin esterase in pectinase, papain in protease, b-amylase saccharifying enzyme and isoamylase in amylase, and cellulase can improve the combustibility of Nanping cigar tobacco leaves, compared with other pectinase, protease, amylase and hemicellulase; compared with the blank group, the enzyme preparation group can improve the combustibility of Nanping cigar tobacco leaves. Therefore, pectin lyase, pectin esterase, papain, b-amylase saccharifying enzyme, isoamylase and cellulase are selected as the compound enzyme preparation.

[0047] Example 3 Influence of different enzymolysis modes on the combustibility of Nanping cigar tobacco leaves (1) Take pectin lyase, pectin esterase, papain, b-amylase, isoamylase and cellulase, respectively added in pure water, and respectively prepared into enzyme preparation solution with mass concentration of 2%. (2) Scheme 1: Mix pectin lyase, pectin esterase and cellulase solution, mix papain, b-amylase and isoamylase solution, put Nanping cigar tobacco leaves in the mixed enzyme preparation solution of pectin lyase, pectin esterase and cellulase, carry out enzymolysis at 35℃ for 30 minutes, take out and place in the mixed enzyme preparation solution of papain, b-amylase and isoamylase after enzyme inactivation, carry out enzymolysis at 35℃ for 30 minutes, take out and dry, and control the moisture content of Nanping cigar tobacco leaves to be 12%.

[0048] Scheme 2: Mix pectin lyase, pectin esterase and cellulase solution, mix papain, b-amylase and isoamylase solution, put Nanping cigar tobacco leaves in the mixed enzyme preparation solution of papain, b-amylase and isoamylase, carry out enzymolysis at 35℃ for 30 minutes, take out and place in the mixed enzyme preparation solution of pectin lyase, pectin esterase and cellulase after enzyme inactivation, carry out enzymolysis at 35℃ for 30 minutes, take out and dry, and control the moisture content of Nanping cigar tobacco leaves to be 12%.

[0049] Scheme 3: Pectin lyase, pectin esterase, papain solution is mixed, β-amylase, isoamylase, cellulase solution is mixed, Nanping cigar tobacco is placed in the mixed enzyme preparation solution of pectin lyase, pectin esterase, papain, and is enzymolyzed at 35℃ for 30 minutes, taken out and dried after enzyme inactivation, placed in the mixed enzyme preparation solution of β-amylase, isoamylase, cellulase, and is enzymolyzed at 35℃ for 30 minutes, taken out and dried, and the moisture content of Nanping cigar tobacco is controlled to be 12%.

[0050] Scheme 4: Pectin lyase, pectin esterase, papain, β-amylase, isoamylase, cellulase solution is mixed, Nanping cigar tobacco is placed in the mixed enzyme preparation solution of pectin lyase, pectin esterase, cellulase, papain, β-amylase, isoamylase, and is enzymolyzed at 35℃ for 30 minutes, taken out and dried, and the moisture content of Nanping cigar tobacco is controlled to be 12%; The burning property of Nanping cigar tobacco is determined according to the method of Example 1, and the specific results are shown in Table 14. Table 14 According to Table 14, under the same enzymolysis time of 30 minutes of each enzyme preparation, scheme 1, i.e., first mixed enzyme solution of pectin lyase, pectin esterase and cellulase, and then mixed enzyme solution of papain, β-amylase and isoamylase, can significantly improve the burning property of Nanping cigar tobacco, and is better than schemes 2-4. The possible reason is that pectin lyase, pectin esterase and cellulase are all wall-breaking enzymes, which can destroy the cell wall of plants and make the internal structure loose, so that papain, β-amylase and isoamylase can more easily combine with their respective substrates, thereby improving the efficiency of enzymolysis.

[0051] Example 4: Effect of different enzymolysis parameters on the burning property of Nanping cigar tobacco 1), Effect of one-time enzymolysis temperature on the burning property of Nanping cigar tobacco (1) Pectin lyase, pectin esterase, papain, β-amylase, isoamylase, cellulase are taken and respectively added into pure water to prepare enzyme preparation solutions with a mass concentration of 2%; (2) Pectin lyase, pectin esterase, cellulase solution is mixed, papain, β-amylase, isoamylase solution is mixed, Nanping cigar tobacco is placed in the mixed enzyme preparation solution of pectin lyase, pectin esterase and cellulase, and is enzymolyzed at (30℃, 35℃, 40℃, 45℃) for 30 minutes, taken out and placed in the mixed enzyme preparation solution of papain, β-amylase and isoamylase after enzyme inactivation, and is enzymolyzed at 35℃ for 30 minutes, taken out and dried, and the moisture content of Nanping cigar tobacco is controlled to be 12%; The specific results are shown in Table 14. Figure 5 Table 14 Figure 5It can be seen that as the temperature of the first enzymatic hydrolysis increases, the smoldering rate (mm / min) and the ash value increase, reaching the maximum value at 40℃. Therefore, the temperature of the first enzymatic hydrolysis is selected as 40℃.

[0052] 2) The effect of single enzymatic hydrolysis time on the combustibility of Nanping cigar tobacco leaves (1) Take pectin lyase, pectin esterase, papain, β-amylase, isoamylase and cellulase, add them to pure water respectively, and prepare enzyme preparation solutions with a mass concentration of 2% respectively; (2) Mix pectin lyase, pectin esterase and cellulase solutions, mix papain, β-amylase and isoamylase solutions, place Nanping cigar tobacco leaves in a mixed enzyme preparation solution of pectin lyase, pectin esterase and cellulase, and enzymatically hydrolyze at 40℃ for (15, 20, 25, 30, 35) minutes. After removing and inactivating the enzymes, place them in a mixed enzyme preparation solution of papain, β-amylase and isoamylase, and enzymatically hydrolyze at 35℃ for 30 minutes. Remove and dry, and control the moisture content of Nanping cigar tobacco leaves to 12%. Specific results are as follows Figure 6 As shown, from Figure 6 It can be seen that as the enzymatic hydrolysis time increases, the smoldering rate (mm / min) and ash value increase, reaching their maximum values ​​at 30 min. Therefore, a single enzymatic hydrolysis time of 30 min is selected.

[0053] 3) The effect of secondary enzymatic hydrolysis temperature on the combustibility of Nanping cigar tobacco leaves (1) Take pectin lyase, pectin esterase, papain, β-amylase, isoamylase and cellulase, add them to pure water respectively, and prepare enzyme preparation solutions with a mass concentration of 2% respectively; (2) Mix pectin lyase, pectin esterase and cellulase solutions, mix papain, β-amylase and isoamylase solutions, place Nanping cigar tobacco leaves in a mixed enzyme preparation solution of pectin lyase, pectin esterase and cellulase, enzymatically hydrolyze at 40℃ for 30 minutes, remove and inactivate the enzyme, place in a mixed enzyme preparation solution of papain, β-amylase and isoamylase, enzymatically hydrolyze at (32℃, 35℃, 38℃, 41℃, 44℃) for 30 minutes, remove and dry, and control the moisture content of Nanping cigar tobacco leaves to 12%; Specific results are as follows Figure 7 As shown, from Figure 7 It can be seen that as the secondary enzymatic hydrolysis temperature increases, the smoldering rate (mm / min) and ash value increase, reaching their maximum values ​​at 38℃. Therefore, the secondary enzymatic hydrolysis temperature is selected as 38℃.

[0054] 4) The effect of secondary enzymatic hydrolysis time on the combustibility of Nanping cigar tobacco leaves (1) Take pectin lyase, pectin esterase, papain, β-amylase, isoamylase and cellulase, add them to pure water respectively, and prepare enzyme preparation solutions with a mass concentration of 2% respectively; (2) Mix pectin lyase, pectin esterase and cellulase solutions, mix papain, β-amylase and isoamylase solutions, place Nanping cigar tobacco leaves in a mixed enzyme preparation solution of pectin lyase, pectin esterase and cellulase, enzymatically hydrolyze at 40℃ for 30 minutes, remove and inactivate the enzyme, place in a mixed enzyme preparation solution of papain, β-amylase and isoamylase, enzymatically hydrolyze at 30℃ for (15, 20, 25, 30, 35) minutes, remove and dry, and control the moisture content of Nanping cigar tobacco leaves to 12%; Specific results are as follows Figure 8 As shown, from Figure 8 It can be seen that as the secondary enzymatic hydrolysis time increases, the smoldering rate (mm / min) and ash value increase, reaching their maximum values ​​at 25 min. Therefore, the secondary enzymatic hydrolysis time is selected as 25 min.

[0055] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. A compound enzyme preparation for improving the combustibility of Nanping cigar tobacco leaves, characterized in that... The compound enzyme preparation includes solutions of pectin lyase, pectin esterase, papain, β-amylase, isoamylase, and cellulase. The enzyme solutions are prepared by adding pectin lyase, pectin esterase, papain, β-amylase, isoamylase, and cellulase to purified water to prepare 2% (w / w) enzyme solutions. The flat cigar tobacco leaves are prepared using the M1LTHH process, meaning the leaves are harvested at a green maturity. The drying process is as follows: withering period: humidity 95-98%RH, temperature 22-25℃; yellowing period: humidity 90-95%RH, temperature 25-28℃; browning period: humidity 85-90%RH, temperature 28-31℃; color fixing period: humidity 70-80%RH, temperature 31-34℃; stalk drying period: humidity 60-70%RH, temperature 31-34℃.

2. A method for controlling the enzymatic hydrolysis of Nanping cigar tobacco leaves using the compound enzyme preparation described in claim 1, characterized in that... Includes the following steps: (1) Take pectin lyase, pectin esterase, papain, β-amylase, isoamylase and cellulase, add them to pure water respectively, and prepare enzyme preparation solutions with a mass concentration of 2% respectively; (2) Mix pectin lyase, pectin esterase and cellulase solutions, and mix papain, β-amylase and isoamylase solutions. Place Nanping cigar tobacco leaves in the mixed enzyme preparation solution of pectin lyase, pectin esterase and cellulase, and enzymatically hydrolyze them once at 35-45℃ for 25-35 minutes. After removing and inactivating the enzymes, place them in the mixed enzyme preparation solution of papain, β-amylase and isoamylase, and enzymatically hydrolyze them a second time at 25-35℃ for 20-30 minutes. Remove and dry them, controlling the moisture content of Nanping cigar tobacco leaves to be 12%.

3. The method for controlling enzymatic hydrolysis of Nanping cigar tobacco leaves using the compound enzyme preparation of claim 1, as described in claim 2, is characterized in that... The temperature for the first enzymatic hydrolysis in step (2) is 35°C.

4. The method for controlling enzymatic hydrolysis of Nanping cigar tobacco leaves using the compound enzyme preparation of claim 1, as described in claim 2, is characterized in that... The temperature for the first enzymatic hydrolysis in step (2) is 40℃.

5. The method for controlling enzymatic hydrolysis of Nanping cigar tobacco leaves using the compound enzyme preparation of claim 1 according to claim 2, characterized in that, The enzymatic hydrolysis time in step (2) is 35 minutes.

6. The method for controlling enzymatic hydrolysis of Nanping cigar tobacco leaves using the compound enzyme preparation of claim 1, as described in claim 2, is characterized in that... The enzymatic hydrolysis time in step (2) is 30 minutes.

7. The method for controlling enzymatic hydrolysis of Nanping cigar tobacco leaves using the compound enzyme preparation of claim 1, as described in claim 2, is characterized in that... The temperature for the secondary enzymatic hydrolysis in step (2) is 35℃.

8. The method for controlling enzymatic hydrolysis of Nanping cigar tobacco leaves using the compound enzyme preparation of claim 1 according to claim 2, characterized in that, The temperature for the secondary enzymatic hydrolysis in step (2) is 30℃.

9. The method for controlling enzymatic hydrolysis of Nanping cigar tobacco leaves using the compound enzyme preparation of claim 1 according to claim 2, characterized in that, The time for the second enzymatic hydrolysis in step (2) is 30 minutes.

10. The method for controlling enzymatic hydrolysis of Nanping cigar tobacco leaves using the compound enzyme preparation of claim 1 according to claim 2, characterized in that, The time for the second enzymatic hydrolysis in step (2) is 25 minutes.

Images