Method for degrading lignin in tobacco stems

Abstract

The invention belongs to the field of biotechnology and discloses a method for degrading lignin in tobacco stems. The method for degrading the lignin in the tobacco stems comprises the steps of mixing the tobacco stems with laccase and water, and performing enzymolysis on the mixture for 36 hours to 60 hours at the temperature of 40 DEG C-48 DEG C at a pH of 4.5 to 5.5. The method for degrading the lignin in tobacco stems is simple in operation, small in damage to the tobacco stems serving as raw materials, and capable of remarkably increasing the degradation rate of the lignin in the tobacco stems, improving quality of cut tobacco stems, and fully utilizing tobacco stem resources. Offensive odor and irritation of cigarettes added with tobacco stem enzymolysis products prepared by the method can be decreased remarkably, lignin odor is weakened greatly, and fragrance and agreeable aftertaste of the cigarettes can be improved.

Description

technical field

[0001] The invention belongs to the field of biotechnology, in particular to a method for degrading lignin in tobacco stems. Background technique

[0002] Tobacco is an annual herbaceous plant of the family Solanaceae, which is mainly used to make shredded tobacco, cigarettes and cigars. Tobacco leaves are the main processing part. Tobacco leaves are separated into sheets and stems after threshing and rebaking. Tobacco stems are the thick and hard veins of tobacco leaves, accounting for about 25%-30% of the leaf weight. They are the main by-products of the tobacco industry. one of the products. Existing methods for utilizing tobacco stems mainly include manufacturing cigarettes, producing tobacco sheets, extracting nicotine and solanesol, and the like. In the process of making cigarettes, some tobacco stems are shredded and expanded and added to cigarette products, which can increase the filling value of shredded tobacco and reduce the content of tar and ca...

Examples

Embodiment 1

[0055] Example 1 Condition screening of laccase enzymatic hydrolysis of lignin in tobacco stems

[0056] Tobacco stem: provided by Zhengzhou Cigarette Factory, dried at 60°C to constant weight, then pulverized with a pulverizer, and passed through a 60-mesh sieve to obtain dry tobacco stem powder for future use.

[0057] Determination of lignin content: Wrap the accurately weighed 1g stalk end sample with filter paper, then put it into a Soxhlet extractor and extract it with a 1:2 ethanol-benzene mixture for 6 hours, take it out and air-dry it, then move it to a 250mL sample with a grinding port Add 15mL of 72% sulfuric acid to a conical flask with a glass stopper, keep it in a constant temperature water bath at 25°C for 2.5h and keep stirring, then transfer it all to a 1000mL conical flask, dilute with water until the concentration of sulfuric acid is 3 %, keep the volume and boil for 4 hours; filter with constant weight quantitative filter paper, then wash until neutral, the...

Embodiment 2

[0085] Example 2 Utilizing laccase to enzymatically hydrolyze lignin in tobacco stems

[0086]Accurately weigh 20g of tobacco stem powder and place it in a triangular flask, add laccase 20000U (purchased from Beijing Novozymes), 900mL of acetic acid-sodium acetate buffer solution, and finally add water to 1000mL to adjust the pH value of the enzymolysis solution 5.0, the Erlenmeyer flask was placed in a full-temperature shaking incubator with a rotation speed of 150r / min and 45°C for enzymatic hydrolysis for 48 hours. After the enzymatic hydrolysis, the precipitate was collected by filtration, washed with 100 mL of water, and dried to obtain 17.02 g of the product. The inactivated enzymatic hydrolyzate was used as a control, and other treatment conditions were the same as above. Measure the content of lignin in the sample before and after enzymolysis according to the lignin content determination method provided in Example 1, the content of lignin in the tobacco stem before en...

Embodiment 3

[0087] Example 3 Degradation of Lignin in Tobacco Stems by Laccase

[0088] Accurately weigh 20g of tobacco stem powder and place it in a triangular flask, add 16000U laccase (purchased from Beijing Novozymes), 700mL of buffer solution, and finally add water to 800mL to adjust the pH value of the enzymolysis solution to 5.5. The Erlenmeyer flask was placed in a full-temperature shaking incubator with a rotating speed of 160r / min, and the enzymatic hydrolysis was carried out at 40°C for 60h. After the enzymatic hydrolysis, the precipitate was collected by filtration, washed with 100 mL of water, and dried to obtain 17.18 g of the product. The inactivated enzymatic hydrolyzate was used as a control, and other treatment conditions were the same as above. Measure the content of lignin in the sample before and after enzymolysis according to the lignin content determination method provided in Example 1, the content of lignin in the tobacco stem before enzymolysis is 4.74%, the cont...

technical field

[0001] The invention belongs to the field of biotechnology, in particular to a method for degrading lignin in tobacco stems. Background technique

[0002] Tobacco is an annual herbaceous plant of the family Solanaceae, which is mainly used to make shredded tobacco, cigarettes and cigars. Tobacco leaves are the main processing part. Tobacco leaves are separated into sheets and stems after threshing and rebaking. Tobacco stems are the thick and hard veins of tobacco leaves, accounting for about 25%-30% of the leaf weight. They are the main by-products of the tobacco industry. one of the products. Existing methods for utilizing tobacco stems mainly include manufacturing cigarettes, producing tobacco sheets, extracting nicotine and solanesol, and the like. In the process of making cigarettes, some tobacco stems are shredded and expanded and added to cigarette products, which can increase the filling value of shredded tobacco and reduce the content of tar and ca...

Examples

Embodiment 1

[0055] Example 1 Condition screening of laccase enzymatic hydrolysis of lignin in tobacco stems

[0056] Tobacco stem: provided by Zhengzhou Cigarette Factory, dried at 60°C to constant weight, then pulverized with a pulverizer, and passed through a 60-mesh sieve to obtain dry tobacco stem powder for future use.

[0057] Determination of lignin content: Wrap the accurately weighed 1g stalk end sample with filter paper, then put it into a Soxhlet extractor and extract it with a 1:2 ethanol-benzene mixture for 6 hours, take it out and air-dry it, then move it to a 250mL sample with a grinding port Add 15mL of 72% sulfuric acid to a conical flask with a glass stopper, keep it in a constant temperature water bath at 25°C for 2.5h and keep stirring, then transfer it all to a 1000mL conical flask, dilute with water until the concentration of sulfuric acid is 3 %, keep the volume and boil for 4 hours; filter with constant weight quantitative filter paper, then wash until neutral, the...

Embodiment 2

[0085] Example 2 Utilizing laccase to enzymatically hydrolyze lignin in tobacco stems

[0086]Accurately weigh 20g of tobacco stem powder and place it in a triangular flask, add laccase 20000U (purchased from Beijing Novozymes), 900mL of acetic acid-sodium acetate buffer solution, and finally add water to 1000mL to adjust the pH value of the enzymolysis solution 5.0, the Erlenmeyer flask was placed in a full-temperature shaking incubator with a rotation speed of 150r / min and 45°C for enzymatic hydrolysis for 48 hours. After the enzymatic hydrolysis, the precipitate was collected by filtration, washed with 100 mL of water, and dried to obtain 17.02 g of the product. The inactivated enzymatic hydrolyzate was used as a control, and other treatment conditions were the same as above. Measure the content of lignin in the sample before and after enzymolysis according to the lignin content determination method provided in Example 1, the content of lignin in the tobacco stem before en...

Embodiment 3

[0087] Example 3 Degradation of Lignin in Tobacco Stems by Laccase

[0088] Accurately weigh 20g of tobacco stem powder and place it in a triangular flask, add 16000U laccase (purchased from Beijing Novozymes), 700mL of buffer solution, and finally add water to 800mL to adjust the pH value of the enzymolysis solution to 5.5. The Erlenmeyer flask was placed in a full-temperature shaking incubator with a rotating speed of 160r / min, and the enzymatic hydrolysis was carried out at 40°C for 60h. After the enzymatic hydrolysis, the precipitate was collected by filtration, washed with 100 mL of water, and dried to obtain 17.18 g of the product. The inactivated enzymatic hydrolyzate was used as a control, and other treatment conditions were the same as above. Measure the content of lignin in the sample before and after enzymolysis according to the lignin content determination method provided in Example 1, the content of lignin in the tobacco stem before enzymolysis is 4.74%, the cont...

Images