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A method for extracting lignin from tobacco leaves by enzymatic hydrolysis

A technology of lignin and enzymatic hydrolysis, applied in the field of lignin extraction, can solve the problems of tobacco leaf pretreatment effect to be improved, low yield, time-consuming and the like, and achieve the effects of avoiding dissolution loss, high yield, simple and efficient operation

Active Publication Date: 2016-03-30
CHINA TOBACCO GUANGDONG IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the above methods are all based on the purpose of increasing the conversion rate of sugar, but ignore the structural changes of lignin and the loss of lignin in the pretreatment process, resulting in low yields and other issues.
In addition, organic solvents are also commonly used to enhance enzymatic extraction of lignin, such as acetone (Journal of pulp and paperscience, 2003, 29(7):235-240) and mixed solvents of benzene / toluene-ethanol / ether (Industrial Crops Products., 2010, 32:551 -559), the above organic solvent pretreatment method is not only time-consuming and dangerous, but also the pretreatment effect on tobacco leaves needs to be improved

Method used

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  • A method for extracting lignin from tobacco leaves by enzymatic hydrolysis
  • A method for extracting lignin from tobacco leaves by enzymatic hydrolysis

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Experimental program
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Effect test

Embodiment 1

[0038]S1. Weigh 10g of tobacco leaf powder (moisture content 94.14%, particle size 60-100 mesh), and disperse the tobacco leaf powder in a mixed solution of 250mL lithium hydroxide, urea and water at -16°C. The mass ratio of lithium hydroxide, urea and water is 7:3:90. After stirring for 40 min under the condition of maintaining -16°C, filter with a 600-mesh filter cloth, and wash the solid with pure water until neutral. Vacuum drying at 50°C for 24 hours yielded 3.6738 g of solid, and the cellulose content of pretreated tobacco leaves was determined to be 19.85% by literature method (NREL / TP-510-42618, Determination of Structural Carbohydrates and Lignin in Biomass. 2008).

[0039] S2. Weigh 1g of the solid obtained from S1, add it to 50mL acetic acid-sodium acetate buffer solution, the pH value is 4.8, and the ionic strength is 50mmol / L, then add 20FPU / g cellulase (CTec2, Novi Letter China Biotechnology Co., Ltd.), react at a temperature of 50 ℃ for 72h. After the reaction...

Embodiment 2

[0044] S1. Weigh 10g of tobacco leaf powder (moisture content 94.14%, particle size 60-100 mesh), and disperse the tobacco leaf powder in a mixed solution of 250ml lithium hydroxide, urea and water at -16°C. The mass ratio of lithium hydroxide, urea and water is 7:2:91. After stirring for 40 minutes under the condition of maintaining -16°C, filter with a 600-mesh filter cloth, and keep the filtrate for UV testing. And wash the solid with pure water until neutral. Vacuum drying at 50°C for 24 hours yielded 3.4737 g of solid, and the cellulose content of the pretreated tobacco leaves was determined to be 19.71%.

[0045] S2. Weigh 1g of the solid obtained from S1, add it to 50mL acetic acid-sodium acetate buffer solution, the pH is 4.8, and the ionic strength is 50mmol / L, then add 20FPU / g cellulase (CTec2, Novozymes provided by China Biotechnology Co., Ltd.), and reacted at 50 °C for 72 h. After the reaction, the hydrolyzate of sugar compounds and crude lignin were obtained b...

Embodiment 3

[0048] S1. Weigh 10 g of dried tobacco leaf powder (moisture content 94.14%, particle size 60-100 mesh), and disperse the tobacco leaf powder in a mixed solution of 250 ml of lithium hydroxide, urea and water at -16°C. The mass ratio of lithium hydroxide, urea and water in the mixed solution is 8:4:88. After stirring for 40 minutes under the condition of maintaining -16°C, filter with a 600-mesh filter cloth, and keep the filtrate A for ultraviolet testing. And wash the solid with pure water until neutral. Vacuum drying at 50°C for 24 hours yielded 3.4923 g of solid, and the cellulose content of the pretreated tobacco leaves was determined to be 19.23%.

[0049] S2. Weigh 1g of the solid obtained from S1, add it into 50mL acetic acid-sodium acetate buffer solution, the pH is 4.8, and the ionic strength is 50mmol / L, then add 10FPU / g cellulase (CTec2, Novozymes provided by China Biotechnology Co., Ltd.), and reacted at 50 °C for 72 h. After the reaction, the hydrolyzate of su...

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Abstract

The invention discloses a method of extracting tobacco leaf lignin by an enhanced enzymolysis process. The method comprises the following steps of dispersing dried tobacco leaf powder at a low temperature into mixed liquor of lithium hydrate, urea and water for pre-treating; and carrying out enzymolysis on the pre-treated tobacco leaf powder by cellulose and purifying acidy dioxane liquor to obtain tobacco leaf lignin. The method disclosed by the invention not only overcomes the defect that the lignin structure is changed by lignin dissolution loss caused by the conventional tobacco leaf lignin extracting method, so that purity of the tobacco leaf lignin is remarkably improved, but also is beneficial to subsequent tobacco lea lignin analysis and research. Reagent adopted by the method disclosed by the invention is non-toxic ad pollution-free, low in cost, simple and efficient to operate.

Description

technical field [0001] The invention relates to the technical field of lignin extraction, in particular to a method for extracting lignin from tobacco leaves through an enhanced enzymatic hydrolysis method. Background technique [0002] With the continuous development of the tobacco industry, smoking and health issues have attracted much attention, reducing tar and harm has become an inevitable trend and research focus of the tobacco industry. Lignin in tobacco leaves is the main source of harmful substances such as polycyclic aromatic hydrocarbons and aromatic amines in tobacco tar. Studies have shown that the thermal cracking products of lignin mainly contain phenolic substances of methoxyl, hydroxyl and alkyl groups (Experimental Research on Rapid Pyrolysis of Lignin. 2005,39(5):710-714). During the smoking and burning process of cigarettes, the lignin in tobacco may be cracked to produce various phenolic substances with cancer-promoting effects. Studying the pyrolysis ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08H7/00
Inventor 孔浩辉黄翼飞
Owner CHINA TOBACCO GUANGDONG IND