阅读说明：本技术 利用秸秆制备香草醛和4-乙烯基愈创木酚的方法 (Method for preparing vanillin and 4-vinyl guaiacol by using straws ) 是由 高旻天 侯蓉蓉 崔收庆 于 2019-09-12 设计创作，主要内容包括：本发明涉及一种利用秸秆制备香草醛和4-乙烯基愈创木酚的方法。该方法的步骤包括：(1)粉碎秸秆粒径小于1mm；(2)利用纤维素酶糖化(1)的秸秆；(3)利用阿魏酸酯酶处理步骤(2)中得到的液体；(4)用枯草芽孢杆菌、青霉、酵母和纤维素酶对(3)中的液体进行转化,制备香草醛和4-乙烯基愈创木酚。秸秆的生物炼制,如秸秆的生物乙醇转化,其主要问题是纤维素酶成本高,如果能够在其转化过程中制备出香草醛和4-乙烯基愈创木酚这种高附加值产品,乙醇的生产成本就可以大幅度降低。(The invention relates to a method for preparing vanillin and 4-vinyl guaiacol by using straws. The method comprises the following steps: (1) the particle size of the crushed straw is less than 1 mm; (2) saccharifying the straw of (1) with cellulase; (3) treating the liquid obtained in the step (2) by using feruloyl esterase; (4) transforming the liquid in (3) with bacillus subtilis, penicillium, yeast and cellulase to prepare vanillin and 4-vinyl guaiacol. The main problem of the bio-refining of the straws, such as the bio-ethanol conversion of the straws, is that the cost of the cellulase is high, and if the high value-added products, such as vanillin and 4-vinyl guaiacol, can be prepared in the conversion process, the production cost of the ethanol can be greatly reduced.)

1. A method for preparing vanillin and 4-vinyl guaiacol by using straws is characterized by comprising the following steps:

a. the particle size of the crushed straw is less than 1 mm;

b. b, saccharifying the straws obtained in the step a by using cellulase to obtain saccharified liquid containing the saccharified straws;

c. inactivating the saccharified liquid obtained in the step b, and treating with feruloyl esterase to obtain a saccharified liquid treated by feruloyl esterase;

d. and (c) converting the saccharified solution obtained in the step c by using microorganisms to obtain vanillin and 4-vinyl guaiacol.

2. The method according to claim 1, wherein the specific steps of step b are: and (b) immersing the crushed straws obtained in the step a in a cellulase solution with the concentration range of 5-20 FPU/g, adjusting the pH value to 3-12, and saccharifying at 25-65 ℃ and 200rpm for 36-48 h to obtain the crushed straws saccharified by cellulase.

3. The method according to claim 1, wherein the specific steps of step c are: adding feruloyl esterase into the saccharified liquid after the cellulase is saccharified; the mass ratio of the crushed straws to the ferulic acid esterase is as follows: 1 g: 10-100 mg; obtaining a saccharified solution treated by feruloyl esterase, inoculating microorganisms accounting for 3-5% of the volume of the saccharified solution into the saccharified solution, and performing biotransformation for 24-48 hours at the temperature of 25-50 ℃ to obtain vanillin and 4-vinylguaiacol; the microorganism refers to penicillium, yeast or bacillus subtilis.

4. A method for preparing vanillin and 4-vinyl guaiacol by using straws is characterized by comprising the following steps:

a. the particle size of the crushed straw is less than 1 mm;

b. b, fermenting, saccharifying and converting the straws obtained in the step a by using penicillium to obtain vanillin and 4-vinyl guaiacol.

5. The method according to claim 3, wherein the specific steps of step b are: and (b) immersing the crushed straws obtained in the step (a) in ultrapure water, inoculating the cultivated penicillium in a volume of 1-5% of that of an ultrapure water system, performing straw fermentation saccharification conversion at a temperature of 25-65 ℃ and a pH value of 3-7 for 24-48 hours, and obtaining products of vanillin and 4-vinyl guaiacol.

6. A method according to any one of claims 1 to 4 wherein the straw is rice, barley, wheat or corn crop straw. .

Technical Field

The invention relates to comprehensive utilization of crop waste straws. In particular to a method for preparing vanillin and 4-vinyl guaiacol by using straws.

Background

Vanillin is considered one of the most important fragrances in the world, obtained from the pod of the orchid called vanilla, and is widely used in the food, beverage, perfume and pharmaceutical industries. The production of vanilla beans was highest in indonesia (2600 tons), followed by madagassica (1946 tons), china (1300 tons), mexico (395 tons), tanga (202 tons) and komoruo (66 tons) as recorded by the united nations grain agriculture organization statistics database. Due to the low yield and high cost of natural vanillin, the importance of using alternatives to produce vanillin is increasing. Chemically synthesized vanillin is mainly produced from fossil hydrocarbons, and the yield of synthesized vanillin is increased year by year because of its low price, but is prohibited because some production processes may cause environmental and health related problems. 4-vinyl guaiacol is a volatile phenol compound, is widely applied to industries such as food, beverage, medicine, spice and the like, and plays an important role in daily life of people. 4-Vinylguaiacol has clove-like spicy fragrance, and is an appreciable flavor component in Laochi beer in Germany and wheat in Belgium. Meanwhile, 4-vinylguaiacol has medicinal value due to its anticancer and antioxidant functions. 4-Vinylguaiacol can be synthesized by chemical synthesis, natural plant extraction and biotransformation. However, 4-vinylguaiacol obtained from natural plants is limited in amount and cannot meet the increasing market demand. Currently, 4-vinylguaiacol is produced by chemical synthesis on the market, but consumers prefer more environmentally friendly and more natural products. Food legislation in europe and the usa also indicates that flavour materials may be prepared by microbial or enzymatic processes. Therefore, the biotransformation of 4-vinylguaiacol and vanillin is an urgent priority.

The traditional mode of treating straws adopts field incineration, and the emission of the straws can generate greenhouse gases and air pollutants, and the greenhouse gases and the air pollutants can aggravate environmental pollution and threaten human health. Straw is a renewable energy source with a wide variety of sources, mainly composed of cellulose, hemicellulose and lignin, and can be biologically converted into useful materials and biofuels, including lactic acid and ethanol. In the process of pretreating and saccharifying the straws by using the cellulase, the cellulase has stronger capacity of hydrolyzing cellulose to release glucose, and the ferulic acid esterase can also release phenolic acid in the saccharification liquid. Phenolic acids can be converted to 4-vinylguaiacol and vanillin. At present, vanillin and 4-vinyl guaiacol in the market are mostly chemically synthesized, and may have certain safety problems. Therefore, there is interest and interest in obtaining these natural products by biological methods.

Disclosure of Invention

The invention aims to improve the production added value in the process of biological refining of straws, and provides a method for preparing vanillin and 4-vinyl guaiacol by straws, so as to reduce the biological refining cost of straws.

In order to achieve the purpose, the technical scheme adopted by the invention is as follows:

a method for preparing vanillin and 4-vinyl guaiacol by using straws is characterized by comprising the following steps:

a. the particle size of the crushed straw is less than 1 mm;

b. b, saccharifying the straws obtained in the step a by using cellulase to obtain saccharified liquid containing the saccharified straws;

c. inactivating the saccharified liquid obtained in the step b, and treating with feruloyl esterase to obtain a saccharified liquid treated by feruloyl esterase;

d. and (c) converting the saccharified solution obtained in the step c by using microorganisms to obtain vanillin and 4-vinyl guaiacol.

The specific steps of the step b are as follows: and (b) immersing the crushed straws obtained in the step a in a cellulase solution with the concentration range of 5-20 FPU/g, adjusting the pH value to 3-12, and saccharifying at 25-65 ℃ and 200rpm for 36-48 h to obtain the crushed straws saccharified by cellulase.

The specific steps of the step c are as follows: adding feruloyl esterase into the saccharified liquid after the cellulase is saccharified; the mass ratio of the crushed straws to the ferulic acid esterase is as follows: 1 g: 10-100 mg; obtaining a saccharified solution treated by feruloyl esterase, inoculating microorganisms accounting for 3-5% of the volume of the saccharified solution into the saccharified solution, and performing biotransformation for 24-48 hours at the temperature of 25-50 ℃ to obtain vanillin and 4-vinylguaiacol; the microorganism refers to penicillium, yeast or bacillus subtilis.

A method for preparing vanillin and 4-vinyl guaiacol by using straws is characterized by comprising the following steps:

a. the particle size of the crushed straw is less than 1 mm;

b. b, fermenting, saccharifying and converting the straws obtained in the step a by using penicillium to obtain vanillin and 4-vinyl guaiacol.

The specific steps of the step b are as follows: and (b) immersing the crushed straws obtained in the step (a) in ultrapure water, inoculating the cultivated penicillium in a volume of 1-5% of that of an ultrapure water system, performing straw fermentation saccharification conversion at a temperature of 25-65 ℃ and a pH value of 3-7 for 24-48 hours, and obtaining products of vanillin and 4-vinyl guaiacol.

The straw is rice, barley, wheat or corn crop straw.

The invention has the beneficial effects that: the main problem of the bio-refining of the straws, such as the bio-ethanol conversion of the straws, is that the cost of the cellulase is high, and if the high value-added products, such as vanillin and 4-vinyl guaiacol, can be prepared in the conversion process, the production cost of the ethanol can be greatly reduced.

Drawings

FIG. 1 is a diagram showing penicillium enzyme production and saccharification; the penicillium can be directly fermented by straws to produce enzyme, saccharify and convert; wherein A is the amount of feruloyl esterase produced by Penicillium (mU/mL); b is the amount of filter paper enzyme activity produced by Penicillium (FPU/mL); c is the amount (g/L) of glucose produced in the enzyme production process of penicillium; d is the amount (g/L) of xylose produced in the enzyme production process of penicillium; e is the amount (mg/g) of vanillic acid produced in the enzyme production process of penicillium; f is the content (mg/g) of vanillin generated in the enzyme production process of penicillium.

Detailed Description

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.