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Method for producing succinic acid through fermentation by utilizing lemna minor

A technology of succinic acid and duckweed, applied in the field of microbial fermentation, can solve the problem of high cost of raw materials, and achieve the effects of reducing production cost, being environmentally friendly, and reducing the amount of nitrogen source added.

Inactive Publication Date: 2015-08-19
GUANGXI ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] The purpose of the present invention is to provide a method for producing succinic acid by fermenting duckweed in order to solve the problem that the raw material cost is too high in the current succinic acid fermentation production. Actinobacillus succigenes is fermented to produce succinic acid, which not only reduces the production cost of succinic acid, but also realizes the recycling and environmental protection of waste water

Method used

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  • Method for producing succinic acid through fermentation by utilizing lemna minor
  • Method for producing succinic acid through fermentation by utilizing lemna minor
  • Method for producing succinic acid through fermentation by utilizing lemna minor

Examples

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Effect test

Embodiment 1

[0024] This embodiment is a specific example of the method for producing succinic acid by fermentation of duckweed according to the present invention, including the following operation steps and control technical conditions: raw materials of duckweed are collected from Xingning District, Nanning City, Guangxi Zhuang Autonomous Region.

[0025] 1. Crushing and sizing of duckweed raw materials: Duckweed dried at 70°C to a constant weight is pulverized, passed through a 60-mesh sieve, and water is added to adjust the total sugar concentration to 30-70g / L to obtain a duckweed mash ;

[0026] 2. Fermentation culture and preparation: adjust the pH value of duckweed mash to 5.5, add medium temperature α-amylase according to 10u / g duckweed powder, hydrolyze at 85℃ for 30min, adjust the pH to 4.5 by cooling, and float at 105u / g Add saccharifying enzyme to the powdered sugar, hydrolyze at 60℃ for 2h, add corn steep liquor 0-20g / L, KH 2 PO 4 2g / L, CaCl 2 0.3g / L, MgCl 2 0.5-1g / L, basic magnesi...

Embodiment 2

[0039] The Actinobacillus succinate-producing strains were connected to the seed culture medium, cultured in an anaerobic incubator or ordinary incubator at 37°C for 16 hours, and then subjected to secondary propagation and culture for 8 hours according to the 5% (V / V) inoculum. The liquid seed is obtained. Put 8% (V / V) inoculum into a 250mL Erlenmeyer flask containing fermentation medium, with a liquid volume of 150mL, add 20g / L corn steep liquor as a nitrogen source, and add 50g / L basic magnesium carbonate Adjust the pH to 6.0-7.0 and place it in an anaerobic incubator for fermentation at 37°C for 60 hours. Anaerobic box culture conditions: anaerobic box (Don Whitley Scientific DG250anaerobic workstation), mixed gas N 2 : CO 2 : H 2 =8.5: 1.0: 0.5, temperature 37°C. The experimental results are shown in Table 1.

[0040] Table 1 The effect of duckweed with different total sugar concentration on the yield of succinic acid

[0041]

[0042] It can be seen from Table 1 that duckw...

Embodiment 3

[0044] According to the method of Actinobacillus succinate-producing fermented duckweed to produce succinic acid in Example 1, the initial total sugar concentration was 65g / L, and different kinds of nitrogen sources were added. The total nitrogen was added at 1.5g / L. The control did not add anything. Nitrogen source, the inoculum amount is 8% (V / V), the 250mL Erlenmeyer flask of the fermentation medium, the liquid volume is 150mL, placed in an anaerobic incubator at 37°C for 60h fermentation, HPLC is used to determine succinic acid and main by-products Content, the results are shown in Table 2.

[0045] Table 2 Effects of different nitrogen sources on the fermentation of duckweed to produce succinic acid

[0046]

[0047]

[0048] It can be seen from Table 2 that the most suitable nitrogen source for succinic acid fermentation is yeast powder, followed by corn steep liquor. When urea is the nitrogen source, the effect is poor (34.77g / L), but even if no nitrogen source is added The...

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Abstract

The invention relates to a method for producing succinic acid through fermentation by utilizing lemna minor. The method comprises the following steps of drying the lemna minor, crushing, and then adding water for slurry mixing; hydrolyzing starch contained in the lemna minor by utilizing an alpha-amylase and a saccharifying enzyme to release fermentable saccharide as a carbon source; adding a certain amount of protease to hydrolyze protein contained in the lemna minor to obtain a material as a nitrogen source; adding needed nutrient substances and a pH regulating agent, and sterilizing to be used as a fermentation culture medium; adding a cultured actinobacillus seed capable of generating succinic acid under a sterile condition, placing into an anaerobic culture box for culture and fermentation to obtain succinic acid. The method for producing succinic acid by utilizing a novel energy plant lemna minor, which is disclosed by the invention, can not only be used for solving the problem of secondary pollution probably caused by large-quantity propagation of the lemna minor in a sewage purifying process, but also provide a low-cost renewable raw material source for the biological refining of succinic acid, is environmentally friendly and can be used for solving the problem of petrochemical resource shortage of the chemical synthesis of succinic acid.

Description

Technical field [0001] The invention belongs to the technical field of microbial fermentation, and specifically relates to a method for producing succinic acid by fermentation of duckweed. Background technique [0002] Succinic acid, also known as succinic acid (succsinic acid), is a common natural organic acid. It is one of the intermediate products of biological TCA cycle, and it is widely present in animals, plants and microorganisms. Succinic acid is an important organic synthesis intermediate. It is recognized as GRAS by the US FDA. It is generally considered safe. It is mainly used in medicine, food, dyes, biodegradable plastics, surfactants, animal feed additives, plant growth stimulators, Fields such as detergents and green solvents. [0003] At present, the production of succinic acid mainly uses a chemical synthesis method, from butane to maleic anhydride (maleic anhydride), and then electrolytic reduction reaction to generate succinic acid. The production of succinic a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/46C12R1/01
CPCC12P7/46
Inventor 申乃坤王青艳廖思明朱绮霞朱婧秦艳
Owner GUANGXI ACAD OF SCI
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