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Method for producing succinic acid by simultaneous saccharification and fermentation of straw raw material

A technology of simultaneous saccharification fermentation and succinic acid, applied in the field of bioengineering, can solve the problem of no succinic acid, etc., and achieve the effects of promoting utilization, good application value, and reducing investment and operating costs

Inactive Publication Date: 2009-12-16
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no research example of synchronous saccharification and fermentation of straw raw materials to produce succinic acid

Method used

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  • Method for producing succinic acid by simultaneous saccharification and fermentation of straw raw material
  • Method for producing succinic acid by simultaneous saccharification and fermentation of straw raw material
  • Method for producing succinic acid by simultaneous saccharification and fermentation of straw raw material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Fermentation of corn stalk enzyme hydrolyzate by dilute alkali pretreatment to produce succinic acid

[0032] Take 80g of dried and pulverized corn stalks, mix them with 1200mL of 1% (w / w) sodium hydroxide solution, treat at 120°C for 1h, filter the residue, wash the residue with water until neutral, and dry at 60°C to obtain The weight loss of 41.3g of treated straw particles was 48.3%. The composition analysis of corn straw before and after dilute alkali treatment is shown in Table 1.

[0033] Table 1 Analysis of corn stalk components before and after dilute alkali treatment

[0034]

[0035] Take 2g of the treated straw particles, put it into a 150mL Erlenmeyer flask, add 42mL pH4.8 citrate buffer, 6mL cellulase solution, add enzyme amount at 20IU / g (substrate), total liquid volume 50mL, Hydrolysis was carried out on a water bath shaker at 50°C for 36 hours, and the sugar content in the hydrolyzed solution was determined as follows:

[0036] Table 2 Co...

Embodiment 2

[0041] Example 2 Synchronous fermentation of corn stalks after dilute alkali pretreatment to produce succinic acid

[0042] Weigh 2g of dry straw particles pretreated by dilute alkali into a 150mL anaerobic bottle, add 50mL of culture solution, which contains: yeast extract 10g / L, sodium dihydrogen phosphate 1.5g / L, dipotassium hydrogen phosphate 1.5g / L , anhydrous calcium chloride 0.2g / L, MgCl 2 0.15g / L, sodium acetate 0.5g / L, adjusted to pH 6.8, stirred evenly, sterilized at 121°C for 15min, then inserted 2.5mL of Actinobacillus succinicum CGMCC1593 anaerobic seeds, and added commercially available Celluclast 1.5L Weixin cellulase, the dosage is 20IU / g substrate. Shake well in 100% CO 2 Under these conditions, the fermentation broth was cultured statically for 72 hours, and the content of succinic acid in the fermentation broth was determined to be 20.8g / L.

Embodiment 3

[0043] Embodiment 3 Trichoderma reesei shake flask liquid fermentation produces cellulase

[0044] Fermentation medium: peptone 0.3%, ammonium sulfate 0.2%, yeast extract 0.05%, KH 2 PO 4 0.4%, CaCl 2 2H 2 0 0.03%, MgSO 4 ·7H 2 O 0.03%, Tween-80 0.02%, microcrystalline cellulose (Avicel) 1%-2.5%, bran 2%-6%. Fermentation conditions: culture temperature 28~30℃, pH 5.5, culture time 5~6 days, shaker rotation speed 180r·min -1 , 250mL Erlenmeyer bottle liquid 50mL.

[0045] Rinse the well-grown Trichoderma reesei viridans CICC40359 slant with 20mL of normal saline, transfer the rinsing solution into a conical flask equipped with glass beads, shake it well, and inoculate it into the fermentation medium at 5% as seed solution, and measure the enzyme after 6 days of fermentation The live results are shown in Table 4.

[0046] Table 4 Enzyme production with different microcrystalline cellulose and bran content

[0047]

[0048] Example 4-9 Synchronous saccharification an...

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Abstract

The invention discloses a method for producing succinic acid by the simultaneous saccharification and fermentation of a straw raw material and belongs to the technical field of bioengineering. In the method, a straw agricultural waste serving as a raw material is synchronously saccharified and fermented by microorganisms to form the succinic acid; or straws are cut, grounded and pretreated by a diluted alkali to form succinic acid under the coactions of cellulose generated by trichoderma reesei and trichoderma viride and actinobacillus succinogenes; or the straws pretreated by the 70g / L diluted alkali forms 35.5g / L succinic acid through anaerobic fermentation at 38 DEG C. The method has the advantages of cheap and non-grain raw material, low equipment investment and operation cost, environment protection and alleviation of the dependency of chemical synthesis on petrochemical resources.

Description

technical field [0001] The invention relates to a method for producing succinic acid by synchronous saccharification and fermentation of microorganisms using agricultural waste straw as a raw material, and belongs to the technical field of bioengineering. Background technique [0002] Succinic acid, also known as succinic acid, is an important C 4 Platform compounds, which are used to synthesize 1,4-butanediol, tetrahydrofuran, adipic acid, γ-butyrolactone, N-methylpyrrolidone and other bulk chemicals, professional chemicals and polybutylene succinate (PBS), etc. Basic raw material for degradable plastics. As a basic raw material of organic chemical industry, succinic acid is widely used in food, medicine, surfactants, detergents, green solvents, biodegradable plastics and other fields. At present, succinic acid is mainly prepared commercially by the chemical method of catalytic hydrogenation of maleic anhydride followed by hydrolysis. Chemical production of succinic acid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/46C12R1/01
Inventor 郑璞孙志浩方林徐岩
Owner JIANGNAN UNIV
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