Method for increasing content of amino acid in tobacco extracting solution

Abstract

The invention relates to the field of tobaccos, in particular to a method for increasing the content of amino acid in a tobacco extracting solution, which comprises the following steps: mixing the tobacco extracting solution with a biological enzyme, fermenting, and inactivating to obtain a target tobacco extracting solution, the biological enzyme comprises multiple of phytoextraction enzyme, hemicellulase, fungal amylase, glucamylase, laccase or protease, the phytoextraction enzyme comprises cellulase, and the protease is acid protease or neutral protease. According to the method, the variety of amino acids in the tobacco extracting solution can be increased to 17, the content of the amino acids is increased by 20% or above, the process is simple, and operability is high.

Description

Technical Field

[0001] The invention relates to the field of tobacco, and in particular to a method for increasing the amino acid content in tobacco extract. Background Art

[0002] The types and contents of amino acids in tobacco leaves play an important role in the taste and flavor of cigarette products. On the one hand, some types of amino acids are indirect precursors of tobacco aroma precursors. For example, phenylalanine is a precursor of benzyl alcohol and phenylethanol, important aroma components of tobacco leaves. On the other hand, amino acids react with sugars to produce browning, namely the Maillard reaction, to produce many important aroma components of tobacco, such as pyrrole and pyrazine compounds, which are very beneficial for improving the smell of cigarettes, increasing the aroma of tobacco, and reducing impurities and irritation.

[0003] The quality of tobacco extract directly affects the taste of paper-made reconstituted tobacco. Currently, paper-made reconstituted tobacco generally has the disadvantages of having a strong irritating taste and a strong woody smell when smoked, and its aroma is also weaker than that of tobacco leaves. How to improve the taste quality of tobacco extract has always been a focus of the industry. Summary of the invention

[0004] In view of the above-mentioned shortcomings of the prior art, the object of the present invention is to provide a method for increasing the amino acid content in tobacco extract to solve the problems in the prior art.

[0005] To achieve the above-mentioned purpose and other related purposes, the present invention provides a method for increasing the amino acid content in tobacco extract, comprising the following steps: mixing the tobacco extract with a biological enzyme and then fermenting it, and inactivating it after fermentation to obtain the target tobacco extract, wherein the biological enzyme comprises a variety of plant enzymes, hemicellulases, fungal amylases, glucoamylases, laccases or proteases, the plant enzyme comprises cellulase, and the protease is an acidic protease.

[0006] In certain embodiments of the present invention, the mixed fermentation of tobacco extract and biological enzymes comprises the following steps:

[0007] 1) mixing tobacco extract with plant enzyme, hemicellulase, fungal amylase and glucoamylase, and fermenting;

[0008] 2) mixing laccase with the fermentation product of step 1) and continuing fermentation;

[0009] 3) Mixing the protease with the fermentation product of step 2) and continuing the fermentation.

[0010] The invention also provides tobacco extract obtained by the preparation method.

[0011] The invention also provides use of the tobacco extract in preparing tobacco sheets.

[0012] The present invention also provides a tobacco sheet, which is prepared by utilizing the tobacco extract.

[0013] As described above, the method for increasing the amino acid content in tobacco extract of the present invention has the following beneficial effects: by increasing the types and contents of amino acids in the extract, on the one hand, some tobacco aroma precursors are increased, for example, the generated phenylalanine is a precursor of benzyl alcohol and phenylethanol, which are important aroma components of tobacco leaves, and on the other hand, the ability of amino acids to react with sugars to brown, i.e., Maillard reaction, is improved to generate many important tobacco aroma components, such as pyrroles and pyrazine compounds, which are very beneficial for improving the smell of tobacco sheets, increasing tobacco aroma, and reducing miscellaneous odor and irritation. This method can increase the types of amino acids in tobacco concentrate to 17, increase the amino acid content by more than 20%, and has a simple process and strong operability, which can provide technical reference for other technicians in the industry. DETAILED DESCRIPTION

[0014] The invention aims at the low types and contents of amino acids in tobacco extract, and adds biological enzymes to the tobacco extract to increase the types and contents of amino acids. By using different biological enzymes, the types and contents of amino acids in the tobacco extract can be regulated, giving the tobacco extract different smoking tastes.

[0015] Based on this, the present invention provides a method for increasing the amino acid content in a tobacco extract, comprising the following steps: mixing the tobacco extract with a biological enzyme and then fermenting it, and inactivating it after fermentation to obtain a target tobacco extract, wherein the biological enzyme comprises a variety of plant enzymes, hemicellulases, fungal amylases, glucoamylases, laccases or proteases, the plant enzyme comprises cellulase, and the protease is an acidic protease.

[0016] The tobacco extract is usually an original extract obtained by soaking or extracting tobacco with a solvent (such as water, alcohol, ketone or other organic solvent), or a concentrated extract obtained by concentrating the original extract.

[0017] In certain embodiments of the present invention, the density of the tobacco extract is 1.15-1.30 g / m 3 The density of the tobacco extract refers to the mass of the tobacco extract per unit volume.

[0018] In certain embodiments of the present invention, the plant enzyme further comprises pectinase, and the mass ratio of the cellulase to the pectinase is 7:3.

[0019] In certain embodiments of the present invention, the tobacco extract is mixed with plant enzymes, hemicellulase, fungal amylase, glucoamylase, laccase and protease and then fermented.

[0020] In certain embodiments of the present invention, tobacco extract is mixed with plant enzymes, hemicellulases, fungal amylases, glucoamylases, laccases or proteases in separate steps and fermented.

[0021] In certain embodiments of the present invention, the mixed fermentation of tobacco extract and biological enzymes comprises the following steps:

[0022] 1) mixing tobacco extract with plant enzyme, hemicellulase, fungal amylase and glucoamylase, and fermenting;

[0023] 2) mixing laccase with the fermentation product of step 1) and continuing fermentation;

[0024] 3) Mixing the protease with the fermentation product of step 2) and continuing the fermentation.

[0025] In certain embodiments of the present invention, in step 1), the mass ratio of the tobacco extract to the plant enzyme is 1 kg: 0.1-0.5 g.

[0026] The mass ratio of the tobacco extract to the plant enzyme is selected from any one of the following: 1kg: 0.1-0.2g, 1kg: 0.2-0.3g, 1kg: 0.3-0.4g, 1kg: 0.4-0.5g.

[0027] In certain embodiments of the present invention, in step 1), the mass ratio of the tobacco extract to the hemicellulase is 1 kg: 0.03-0.25 g.

[0028] The mass ratio of the tobacco extract to hemicellulase is selected from any one of the following: 1kg: 0.03-0.05g, 1kg: 0.05-0.1g, 1kg: 0.1-0.15g, 1kg: 0.15-0.2g, 1kg: 0.2-0.25g.

[0029] In certain embodiments of the present invention, in step 1), the mass ratio of the tobacco extract to the fungal amylase is 1 kg: 0.3-0.8 g.

[0030] The mass ratio of the tobacco extract to the fungal amylase is selected from any one of the following: 1kg: 0.3-0.4g, 1kg: 0.4-0.5g, 1kg: 0.5-0.6g, 1kg: 0.6-0.7g, 1kg: 0.7-0.8g.

[0031] In certain embodiments of the present invention, in step 1), the mass ratio of the tobacco extract to glucoamylase is 1 kg: 0.05-0.4 g.

[0032] The mass ratio of the tobacco extract to glucoamylase is selected from any of the following: 1kg: 0.05-0.1g, 1kg: 0.1-0.15g, 1kg: 0.15-0.2g, 1kg: 0.2-0.3g, 1kg: 0.3-0.4g.

[0033] In certain embodiments of the present invention, the fermentation time in step 1) is 1.5 to 4 hours, and / or the fermentation temperature is 40° C.-55° C.

[0034] In certain embodiments of the present invention, in step 2), the mass ratio of the tobacco extract to laccase is 1 kg: 0.3-0.8 g.

[0035] The mass ratio of the tobacco extract to laccase is selected from any one of the following: 1kg: 0.3-0.4g, 1kg: 0.4-0.5g, 1kg: 0.5-0.6g, 1kg: 0.6-0.7g, 1kg: 0.7-0.8g.

[0036] In certain embodiments of the present invention, the fermentation time in step 2) is 30-50 min, and / or the fermentation temperature is 40°C-50°C.

[0037] In certain embodiments of the present invention, in step 3), the mass ratio of the tobacco extract to the protease is 1 kg: 0.05-0.4 g.

[0038] The mass ratio of the tobacco extract to the protease is selected from any one of the following: 1kg: 0.05-0.1g, 1kg: 0.1-0.15g, 1kg: 0.15-0.2g, 1kg: 0.2-0.3g, 1kg: 0.3-0.4g.

[0039] In certain embodiments of the present invention, the fermentation time in step 3) is 1-3 hours, and / or the fermentation temperature is 40°C-45°C.

[0040] In certain embodiments of the present invention, the inactivation method is heating at 80-100° C. for 20-40 min.

[0041] The invention also provides tobacco extract obtained by the preparation method.

[0042] The tobacco extract has higher types and contents of amino acids than unfermented tobacco extract.

[0043] The invention also provides use of the tobacco extract in preparing tobacco sheets.

[0044] The present invention also provides a tobacco sheet, which is prepared by utilizing the tobacco extract.

[0045] The following describes the embodiments of the present invention through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention.

[0046] Before further describing the specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terms used in the examples of the present invention are for describing the specific embodiments rather than for limiting the scope of protection of the present invention; in the present specification and claims, unless otherwise expressly stated herein, the singular forms "a", "an" and "the" include plural forms.

[0047] When the embodiments give numerical ranges, it should be understood that, unless otherwise specified in the present invention, both endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those generally understood by those skilled in the art. In addition to the specific methods, equipment, and materials used in the embodiments, according to the grasp of the prior art by those skilled in the art and the record of the present invention, any methods, equipment, and materials of the prior art similar or equivalent to the methods, equipment, and materials described in the embodiments of the present invention can also be used to realize the present invention.

[0048] All the biological enzymes in the following examples are products of Xiasheng Biotechnology Co., Ltd.:

[0049] Phytase, product number SPE-017;

[0050] Hemicellulase product number, FDG-2255;

[0051] Fungal amylase, product number FDY-2247;

[0052] Glucoamylase, product number SDG2413;

[0053] Protease (for tobacco), FFG-0604;

[0054] Laccase, Catalog No. FDY-2204.

[0055] Example 1

[0056] (1) The tobacco raw material is mixed with hot water at 50-70°C in a mass ratio of 1:6 and extracted for 40 minutes. After the extraction, the material is mechanically squeezed to separate the solid and liquid, the solid phase is pulped and paper-made to obtain a tobacco sheet, and the aqueous phase is concentrated to obtain a tobacco extract with a density of 1.15 g / cm 3 ;

[0057] (2) Weigh 1 kg of tobacco extract, density 1.15 g / cm 3 .

[0058] (3) Based on the weight of 1 kg of extract, in the first step, 0.2 g of plant enzyme, 0.05 g of hemicellulase, 0.4 g of fungal amylase, and 0.1 g of glucoamylase were added; the fermentation temperature was 40° C. and the fermentation time was 2 h.

[0059] (4) In the second step, 0.4 g of laccase was added, the fermentation temperature was 40 °C, and the fermentation time was 30 min.

[0060] (5) In the third step, 0.1 g of acidic protease was added, the fermentation temperature was 40°C, and the fermentation time was 1 h.

[0061] (6) The fermented extract was inactivated by heating at 80°C for 20 min to obtain the target extract.

[0062] The amino acid content in the extract was detected by liquid chromatography-mass spectrometry GB 5009.124-2016 method.

[0063] Experimental data

[0064]

[0065] Compared with the unfermented tobacco extract, the types of amino acids in the extract after adding bio-enzyme fermentation increased to 17, and the amino acid content increased by 38.14%.

[0066] Comparative Example 1

[0067] (1) Weigh 1 kg of the tobacco extract prepared in step 1) of Example 1, and first add 0.2 g of pectinase, 0.05 g of hemicellulase, 0.4 g of mesophilic amylase, and 0.1 g of glucoamylase; fermentation temperature is 40° C., and fermentation time is 2 h.

[0068] (2) In the second step, 0.4 g of high temperature desizing enzyme was added, the fermentation temperature was 40°C, and the fermentation time was 30 min.

[0069] (3) In the third step, 0.1 g of acidic protease was added, the fermentation temperature was 40°C, and the fermentation was carried out for 1 h.

[0070] (4) The fermented extract was inactivated by heating at 80°C for 20 min to obtain the target extract.

[0071] The amino acid content in the extract was detected by liquid chromatography-mass spectrometry GB 5009.124-2016 method.

[0072] Experimental data

[0073]

[0074] Compared with the unfermented tobacco extract, the types of amino acids in the extract after adding bio-enzyme fermentation increased to 12, and the amino acid content increased by 9.40%, which was less than that in Example 1.

[0075] Comparative Example 2

[0076] (1) Weigh 1 kg of the tobacco extract prepared in step 1) of Example 1, add 0.1 g of acid protease, ferment at 40° C., and ferment for 1 h.

[0077] (2) In the second step, 0.4 g of laccase was added, the fermentation temperature was 40°C, and the fermentation time was 30 min.

[0078] (3) In the third step, 0.2 g of plant extract enzyme, 0.05 g of hemicellulase, 0.4 g of fungal amylase, and 0.1 g of glucoamylase were added;

[0079] Fermentation temperature is 40℃, and fermentation time is 2h.

[0080] (4) The fermented extract was inactivated by heating at 80°C for 20 min to obtain the target extract.

[0081] The amino acid content in the extract was detected by liquid chromatography-mass spectrometry GB 5009.124-2016 method.

[0082] Experimental data

[0083]

[0084]

[0085] Compared with the unfermented tobacco extract, the number of amino acids in the extract after adding the bio-enzyme fermentation increased to 15, and the amino acid content increased by 5.12%. It can be seen that compared with Example 1, the treatment effect on the extract is reduced after the enzyme addition order is changed.

[0086] Embodiment 2:

[0087] (1) Prepare tobacco extract in the same manner as step 1) of Example 1. Weigh 1 kg of tobacco extract with a density of 1.20 g / cm 3 .

[0088] (2) Based on the weight of 1 kg of extract, in the first step, 0.3 g of plant enzyme, 0.1 g of hemicellulase, 0.5 g of fungal amylase, and 0.2 g of glucoamylase were added; the fermentation temperature was 45° C. and the fermentation was carried out for 3 h.

[0089] (3) In the second step, 0.5 g of laccase was added, the fermentation temperature was 45°C, and the fermentation time was 40 min.

[0090] (4) In the third step, 0.2 g of protease was added, the fermentation temperature was 42° C., and the fermentation was carried out for 2 h.

[0091] (5) The fermented extract was inactivated by heating at 90°C for 30 min to obtain the target extract.

[0092] Experimental data

[0093]

[0094]

[0095] Compared with the unfermented tobacco extract, the types of amino acids in the extract after adding bio-enzyme fermentation increased to 17, and the amino acid content increased by 29.89%.

[0096] Comparative Example 3

[0097] (1) Weigh 1 kg of the tobacco extract prepared in step 1) of Example 2, with a density of 1.20 g / cm 3 .

[0098] (2) Based on the weight of 1 kg of the extract, 0.2 g of acid protease was added in the first step, the fermentation temperature was 42°C, and the fermentation was carried out for 2 h.

[0099] (3) In the second step, 0.5 g of laccase was added, the fermentation temperature was 45°C, and the fermentation time was 40 min.

[0100] (4) In the third step, 0.3 g of plant extractase, 0.1 g of hemicellulase, 0.5 g of fungal amylase, and 0.2 g of glucoamylase were added;

[0101] Fermentation temperature is 45℃, and fermentation time is 3h.

[0102] (5) The fermented extract was inactivated by heating at 90°C for 30 min to obtain the target extract.

[0103] The amino acid content in the extract was detected by liquid chromatography-mass spectrometry GB 5009.124-2016 method.

[0104] Experimental data

[0105]

[0106]

[0107] Compared with the unfermented tobacco extract, the number of amino acids in the extract after adding biological enzyme fermentation increased to 13, and the amino acid content increased by 5.82%. It can be seen that the treatment effect on the extract will decrease after the enzyme addition order is changed.

[0108] Embodiment 3:

[0109] (1) Prepare tobacco extract in the same manner as step 1) of Example 1. Weigh 1 kg of tobacco extract with a density of 1.30 g / cm 3 .

[0110] (2) Based on the weight of 1 kg of extract, in the first step, 0.4 g of plant enzyme, 0.2 g of hemicellulase, 0.7 g of fungal amylase, and 0.3 g of glucoamylase were added; the fermentation temperature was 45° C. and the fermentation was carried out for 4 h.

[0111] (3) In the second step, 0.7 g of laccase was added, the fermentation temperature was 50°C, and the fermentation time was 50 min.

[0112] (4) In the third step, 0.3 g of protease was added, the fermentation temperature was 45°C, and the fermentation was carried out for 3 h.

[0113] (5) The fermented extract was inactivated by heating at 100°C for 40 min to obtain the target extract.

[0114] Experimental data

[0115]

[0116]

[0117] Compared with the unfermented tobacco extract, the types of amino acids in the extract after adding bio-enzyme fermentation increased to 17, and the amino acid content increased by 36.02%.

[0118] Comparative Example 4

[0119] (1) Weigh 1 kg of tobacco extract prepared in step 1) of Example 3, with a density of 1.30 g / cm 3 .

[0120] (2) Based on 1 kg of the extract, in the first step, 0.2 g of chlorophyllase, 0.05 g of nicotine dehydrogenase, and 0.1 g of glucoamylase were added; the fermentation temperature was 45° C. and the fermentation was carried out for 4 h.

[0121] (3) In the second step, 0.4 g of cellulase and 0.3 g of neutral protease were added, the fermentation temperature was 50° C., and the fermentation time was 50 min.

[0122] (4) In the third step, 0.1 g of pectinase was added, the fermentation temperature was 45°C, and the fermentation was carried out for 3 h.

[0123] (5) The fermented extract was inactivated by heating at 100°C for 40 min to obtain the target extract.

[0124] The amino acid content in the extract was detected by liquid chromatography-mass spectrometry GB 5009.124-2016 method.

[0125] Experimental data

[0126]

[0127]

[0128] Compared with the unfermented tobacco extract, the number of amino acid species in the extract after adding bio-enzyme fermentation increased to 13, and the amino acid content increased by 11.34%. It can be seen that compared with the bio-enzyme species in Example 3, the bio-enzyme species in the comparative example have poor treatment effects.

[0129] Example 4 Preparation of tobacco sheet products

[0130] The tobacco extract prepared in Example 3 and Comparative Example 4 is evenly coated on a tobacco sheet base, and then dried to obtain a finished reconstituted tobacco leaf. The specific steps are as follows:

[0131] A. Calculate the required amount of coating liquid according to the coating rate of 36%-38%, coating rate = (weight of product after coating - weight of tobacco base paper before coating) / weight of product after coating * 100%. For example, a coating rate of 36% means that 100g of the coated product contains 36g of tobacco extract in absolute dry weight);

[0132] B. Using a roller coater, evenly coat the tobacco extract onto the tobacco sheet base;

[0133] C. After coating, the product is dried to remove moisture, and a tobacco sheet product is obtained, the moisture content of the product is about 12%.

[0134] The aroma components of the prepared tobacco sheets were detected. The aroma component content of Example 4 was increased by 46.4% compared with that of Comparative Example 4.

[0135] Experimental data

[0136]

[0137]

[0138] Comparative data

[0139]

[0140] Due to the increased content of aroma components in the tobacco flakes, the irritation and paper smell of the tobacco flakes are reduced during the smoking process, and at the same time the aroma quality of the tobacco flakes is improved, thereby comprehensively improving the quality of the tobacco flakes.

[0141] The above examples are for the purpose of illustrating the embodiments disclosed by the present invention and are not to be construed as limiting the present invention. In addition, the various modifications listed herein and the variations of the methods in the invention are obvious to those skilled in the art without departing from the scope and spirit of the present invention. Although the present invention has been specifically described in conjunction with various specific preferred embodiments of the present invention, it should be understood that the present invention should not be limited to these specific embodiments. In fact, various modifications as described above that are obvious to those skilled in the art to obtain the invention should be included within the scope of the present invention.

Claims

1. A method for increasing the amino acid content in tobacco extract, characterized in that: The method comprises the following steps: mixing tobacco extract with biological enzymes and then fermenting them, and then inactivating them after fermentation to obtain target tobacco extract, wherein the biological enzymes comprise plant enzymes, hemicellulase, fungal amylase, glucoamylase, laccase or protease, wherein the plant enzymes comprise cellulase, and the protease is acidic protease.

2. The method according to claim 1, characterized in that The tobacco extract is mixed with plant enzymes, hemicellulase, fungal amylase, glucoamylase, laccase and protease and then fermented; preferably, the tobacco extract is mixed with plant enzymes, hemicellulase, fungal amylase, glucoamylase, laccase or protease in steps and then fermented.

3. The method according to claim 1, characterized in that The mixed fermentation of tobacco extract and biological enzymes includes the following steps: 1) mixing tobacco extract with plant enzyme, hemicellulase, fungal amylase and glucoamylase, and fermenting; 2) mixing laccase with the fermentation product of step 1) and continuing fermentation; 3) Mixing the protease with the fermentation product of step 2) and continuing the fermentation.

4. The method according to claim 1, characterized in that: Step 1) further includes any one or more of the following features: 1A) the mass ratio of the tobacco extract to the plant extract enzyme is 1kg: 0.1-0.5g; 1B) the mass ratio of the tobacco extract to the hemicellulase is 1kg: 0.03-0.25g; 1C) the mass ratio of the tobacco extract to the fungal amylase is 1kg:0.3-0.8g; 1D) The mass ratio of the tobacco extract to glucoamylase is 1kg:0.05-0.4g. 1E) The fermentation time in step 1) is 1.5 to 4 hours, and / or the fermentation temperature is 40°C to 55°C.

5. The method according to claim 1, characterized in that Step 2) further includes any one or more of the following features: 2A) the mass ratio of the tobacco extract to the laccase is 1 kg: 0.3-0.8 g; 2B) Fermentation time is 30-50min; 2C) The fermentation temperature is 40°C-50°C.

6. The method according to claim 1, characterized in that Step 3) further includes any one or more of the following features: 3A) the mass ratio of the tobacco extract to the protease is 1 kg: 0.05-0.4 g; 3B) Fermentation time is 1-3 hours; 3C) The fermentation temperature is 40°C-45°C.

7. The method according to claim 1, characterized in that The inactivation method is heating at 80-100°C for 20-40 minutes.

8. The tobacco extract obtained by the preparation method according to any one of claims 1 to 7.

9. Use of the tobacco extract according to claim 8 in preparing tobacco sheets.

10. A tobacco sheet, wherein the tobacco sheet is prepared using the tobacco extract according to claim 8.

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