Method for producing butane diacid by fermenting Jerusalem artichoke raw material

Abstract

The invention provides a method for producing butane diacid by fermenting cheap Jerusalem artichoke raw material. The invention also provides the application of the Jerusalem artichoke raw material in preparing butane diacid. Meanwhile, the invention provides the application of Actinobacillus succinogenes in the method for preparing the butane diacid by using the Jerusalem artichoke as raw material. The invention has the advantages of using renewable material, being environment-friendly, relieving the lack of the petrification resource in synthesizing the butane diacid by chemical process, and obvious embodiment of no competition with human for grains and no competition with grains for land; therefore, not only the cost of the culture medium of fermentation is reduced, but also the shortage of the grain as fermentation raw material is relieved; moreover, the development and use of large areas of marginal lands such as wasteland, desert and mudflat and the like are solved, thereby being favorable to environment and agriculture and rural construction, and increasing the income of farmers.

Description

technical field

[0001] The invention belongs to the technical field of bioengineering. Specifically, the present invention relates to a method for producing succinic acid by fermenting cheap Jerusalem artichoke raw materials. The invention also relates to the application of Jerusalem artichoke raw material in the preparation of succinic acid. The invention also relates to the application of Actinobacillus succinogenes in the method for preparing succinic acid from Jerusalem artichoke. Background technique

[0002] Succinic acid, also known as succinic acid, has the molecular formula C 4 h 6 o 4 , with a molecular weight of 118.09, is a common natural organic acid that widely exists in the human body, animals, plants and microorganisms.

[0003] Succinic acid is an important four-carbon compound in industry. It is widely used in food, medicine, pesticides, dyes, spices, paints, plastics and material industries as organic synthesis raw materials, intermediate products or ...

Examples

Embodiment 1

[0067] Fermentation and preparation of black koji inulinase

[0068] Aspergillus niger (Apergillus niger) CGMCC 3.4309 was purchased from China General Microorganism Culture Collection and Management Center. The spores were cultivated on the slant of conventional mold potato agar medium, and the spore suspension was prepared (10 6 individual / mL), and were inserted into a 250mL Erlenmeyer flask filled with 80mL enzyme-producing medium. Enzyme production fermentation medium formula (per L): inulin 50g / L, peptone 40g / L, sucrose ester 6g / L, pH6.0. Under the conditions of 30° C. and 200 revolutions / minute, shake the flask to ferment for 2-5 days, respectively take the fermented liquids of different culture days, and measure the enzyme activity. And under the condition of 50 ℃, hydrolysis containing total sugar is 8.5% Jerusalem artichoke slurry (enzyme amount 40U / g total sugar) to measure reducing sugar and calculation hydrolysis rate after 4 hours, 8 hours, 12 hours, the results...

Embodiment 2

[0073] Succinic acid fermentation of Jerusalem artichoke hydrolyzed sugar prepared by different hydrolysis methods

[0074] Seed medium (per L): glucose 5g, yeast extract 5g, K 2 HPO 4 ·3H 2 O 1.0g, NaH 2 PO 4 2H 2 O 1.0g, pH7.0. Anaerobic culture at 37°C for 16 hours.

[0075] Fermentation medium (per L): Jerusalem artichoke hydrolyzed syrup (total reducing sugar) 60g, yeast extract 15g, corn steep liquor 20g, MgCl 2 0.2g, Na 2 HPO 4 12H 2 O 1.5g, NaH 2 PO 4 2H 2 O1.5g, pH 6.5. Anaerobic cultivation at 37°C for 48 hours.

[0076] The method that enzyme hydrolysis Jerusalem artichoke syrup preparation adopts is briefly described as follows:

[0077] (1) Enzymatic hydrolysis: wash the fresh tubers of Jerusalem artichoke, remove the mud, roughly cut to the size of 20-30mm, beat with a tissue masher, and be mixed with a Jerusalem artichoke slurry containing 5%-10% of total sugar, using Example 1 Prepared inulinase (44.1U / ml, 40U / g total sugar for input total sugar)...

Embodiment 3

[0084] Succinic Acid Fermentation of Jerusalem Artichoke Syrup with Different Initial Reducing Sugar Concentrations

[0085] According to the method of Example 2, the Jerusalem artichoke fresh tuber hydrolyzed syrup prepared by using the inulinase hydrolysis of Example 1 was fermented in batches with anaerobic bottles, and when the initial reducing sugar concentration was 30, 45, 55, 65, and 80 g / L respectively, The results are shown in Table 3:

[0086] Table 3 Results of succinic acid fermentation of Jerusalem artichoke syrup with different initial reducing sugar concentrations (anaerobic bottle batch fermentation)

[0087] initial reducing sugar concentration

[0088] As can be seen from Table 3, when the initial reducing sugar concentration is 65-80g / L, the output of succinic acid is the highest, reaching 46.54-46.61g / L. The sugar yield was 79.1%, and the sugar utilization rate was 90.5%.

technical field

[0001] The invention belongs to the technical field of bioengineering. Specifically, the present invention relates to a method for producing succinic acid by fermenting cheap Jerusalem artichoke raw materials. The invention also relates to the application of Jerusalem artichoke raw material in the preparation of succinic acid. The invention also relates to the application of Actinobacillus succinogenes in the method for preparing succinic acid from Jerusalem artichoke. Background technique

[0002] Succinic acid, also known as succinic acid, has the molecular formula C 4 h 6 o 4 , with a molecular weight of 118.09, is a common natural organic acid that widely exists in the human body, animals, plants and microorganisms.

[0003] Succinic acid is an important four-carbon compound in industry. It is widely used in food, medicine, pesticides, dyes, spices, paints, plastics and material industries as organic synthesis raw materials, intermediate products or ...

Examples

Embodiment 1

[0067] Fermentation and preparation of black koji inulinase

[0068] Aspergillus niger (Apergillus niger) CGMCC 3.4309 was purchased from China General Microorganism Culture Collection and Management Center. The spores were cultivated on the slant of conventional mold potato agar medium, and the spore suspension was prepared (10 6 individual / mL), and were inserted into a 250mL Erlenmeyer flask filled with 80mL enzyme-producing medium. Enzyme production fermentation medium formula (per L): inulin 50g / L, peptone 40g / L, sucrose ester 6g / L, pH6.0. Under the conditions of 30° C. and 200 revolutions / minute, shake the flask to ferment for 2-5 days, respectively take the fermented liquids of different culture days, and measure the enzyme activity. And under the condition of 50 ℃, hydrolysis containing total sugar is 8.5% Jerusalem artichoke slurry (enzyme amount 40U / g total sugar) to measure reducing sugar and calculation hydrolysis rate after 4 hours, 8 hours, 12 hours, the results...

Embodiment 2

[0073] Succinic acid fermentation of Jerusalem artichoke hydrolyzed sugar prepared by different hydrolysis methods

[0074] Seed medium (per L): glucose 5g, yeast extract 5g, K 2 HPO 4 ·3H 2 O 1.0g, NaH 2 PO 4 2H 2 O 1.0g, pH7.0. Anaerobic culture at 37°C for 16 hours.

[0075] Fermentation medium (per L): Jerusalem artichoke hydrolyzed syrup (total reducing sugar) 60g, yeast extract 15g, corn steep liquor 20g, MgCl 2 0.2g, Na 2 HPO 4 12H 2 O 1.5g, NaH 2 PO 4 2H 2 O1.5g, pH 6.5. Anaerobic cultivation at 37°C for 48 hours.

[0076] The method that enzyme hydrolysis Jerusalem artichoke syrup preparation adopts is briefly described as follows:

[0077] (1) Enzymatic hydrolysis: wash the fresh tubers of Jerusalem artichoke, remove the mud, roughly cut to the size of 20-30mm, beat with a tissue masher, and be mixed with a Jerusalem artichoke slurry containing 5%-10% of total sugar, using Example 1 Prepared inulinase (44.1U / ml, 40U / g total sugar for input total sugar)...

Embodiment 3

[0084] Succinic Acid Fermentation of Jerusalem Artichoke Syrup with Different Initial Reducing Sugar Concentrations

[0085] According to the method of Example 2, the Jerusalem artichoke fresh tuber hydrolyzed syrup prepared by using the inulinase hydrolysis of Example 1 was fermented in batches with anaerobic bottles, and when the initial reducing sugar concentration was 30, 45, 55, 65, and 80 g / L respectively, The results are shown in Table 3:

[0086] Table 3 Results of succinic acid fermentation of Jerusalem artichoke syrup with different initial reducing sugar concentrations (anaerobic bottle batch fermentation)

[0087] initial reducing sugar concentration

[0088] As can be seen from Table 3, when the initial reducing sugar concentration is 65-80g / L, the output of succinic acid is the highest, reaching 46.54-46.61g / L. The sugar yield was 79.1%, and the sugar utilization rate was 90.5%.

Images