Method for fermentation production of succinic acid by using cotton fiber material to fix actinobacillus succinogenes

An actinomycetes, cotton fiber technology, applied in microorganism-based methods, biochemical equipment and methods, fixed on/in organic carriers, etc., can solve the problem of high production cost, prolonged, low bacterial biomass, etc. problem, to achieve the effect of high acid yield, high conversion rate and high production intensity

Active Publication Date: 2013-12-11
SHANDONG XINGQIANG CHEM IND TECH RES INST CO LTD
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Problems solved by technology

However, at present, this production method is also restricted by the relatively high production cost.
The production of succinic acid by microbial fermentation still has the problems of low bacterial biomass, high substrate concentration and high product concentration in the fermentation process, and the problem that ideal production intensity and conversion rate cannot be obtained at the same time. Therefore, how to obtain the existing high High product concentration and high production intensity and conversion rate have become a research hotspot in the production of succinic acid by fermentation of Actinobacillus succinic acid
[0003] In 2004, U.S. Urbanance reported using plastic composite matrix as immobilized material, installed on the stirring paddle of fermenter, and operated continuously. The production intensity of succinic acid reached 2.1g / L / h, and the conversion rate was 0.72g / g. , but the maximum concentration of succinic acid is only 10.4g / L; they also tried to use the biofilm cycle fermentation system, adding a porous mesh material in the fermenter to enrich the bacteria, the conversion rate and the concentration of succinic acid increased to 0.88g / g and 34g / L, but the production intensity is only 0.9g / L / h
In 2009, South Korea’s Kim et al. fixed cells in a biofilm system for cyclic continuous fermentation. Under the condition of a dilution rate of 0.2 / h, the highest production intensity was 6.63g / L / h, but the concentration of succinic acid was still only 13.26g / L
In 2011, Li Qiang, Institute of Process Technology, Chinese Academy of Sciences, etc. proposed a device for coupling fermentation and extraction, coupling the fermentation reactor with an enlarged adsorption bed. Add the culture medium back into the fermenter for fermentation. When the resin in the adsorption bed is saturated, the entire fermentation cycle ends, and the concentration of succinic acid can be significantly increased (up to 145.2g / L), but the fermentation time is extended from 48h to 126h. The production intensity is 1.3g / L / h, and the average conversion rate is only 0.52g / g
In 2012, Van Heerden and Nicol in South Africa used perlite as a fixed cell material, and perlite was filled in the reactor as an adsorption material for continuous fermentation. When the dilution rate was 0.56 / h, the production intensity of the reactor reached 6.35g / L / h , but the conversion rate is only 0.65g / g
Although some of the above methods effectively improve the production intensity of succinic acid, and some increase the product concentration, there are few methods for obtaining high acid concentration, high conversion rate and high production intensity at the same time

Method used

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  • Method for fermentation production of succinic acid by using cotton fiber material to fix actinobacillus succinogenes
  • Method for fermentation production of succinic acid by using cotton fiber material to fix actinobacillus succinogenes
  • Method for fermentation production of succinic acid by using cotton fiber material to fix actinobacillus succinogenes

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1 batch fermentation

[0023] In the 3L fermenter fiber bed reactor device: Take the fiber bed carrier (the thickness of the fabric is 1mm, the porosity > 95%), and the surface area is 60×6cm 2 , wind it on the stirring shaft of the fermenter, fix the top with a stirring paddle, and fix it with stainless steel wire mesh. The diameter of the wire mesh is about 2.5×2.5cm 2 , and does not affect stirring. The diameter of the fiber bed is about 11cm, the height is 16cm, 115-121°C for 20 minutes, and then 121°C for 20 minutes after overnight;

[0024] The seeds of A. succinogenes CCTCC NO:M2012036 were cultured statically in an anaerobic incubator for 12 hours at a temperature of 38°C. Put the seed medium into the fiber bed reactor, add the seeds of Actinobacillus succinicum shake flask according to the inoculum amount of 2.5%, and incubate at 38°C under the flow of N 2 with CO 2 Under the anaerobic condition of the mixed gas, the adsorption was cultivated for...

Embodiment 2

[0028] Embodiment 2 Batch fermentation repeatedly

[0029] Referring to the method of Example 1, the adsorbed Actinobacillus succinici was cultivated in a 3L fiber bed reactor for repeated batch fermentation. The initial sugar concentration of the fermentation medium was 50g / L glucose, and samples were taken every 3-4 hours to measure the acid production. After an average of 12 hours of fermentation, the glucose concentration dropped to 3g / L. The fresh medium was replaced, and 10 batches of batch fermentation were repeated. The results are shown in Table 2. After the end of the 10th batch, no cell growth was found to stop, which indicated that the fiber bed reactor could still be used.

[0030] Table 2 Fiber bed reactor immobilized repeated batch fermentation to produce succinic acid

[0031]

Embodiment 3

[0032] Embodiment 3 Fed-batch fermentation

[0033] Referring to the method of Example 1, the 3L fiber bed reactor was cultivated in the fiber bed reactor. Actinobacillus succinicum was cultivated, and fed-batch fermentation was carried out in order to obtain a high concentration of succinic acid. The initial glucose of the fermentation medium was 60g / L, and when the residual sugar decreased to 12.4g / L in the first 12 hours of the fermentation process, the glucose concentration was added to 67g / L; after 38 hours, the residual sugar decreased to 5g / L, and the concentration of succinic acid was 87g / L, the conversion rate is 0.83g / g, and the production intensity is 2.38g / L / h.

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Abstract

The invention discloses a method for fermentation production of succinic acid by using a cotton fiber material to fix actinobacillus succinogenes, belonging to the fermentation engineering field. The actinobacillus succinogenes is placed in a fermentation tank which contains a cotton fiber carrier for culturing, adsorbing and fixing; the fermentation cell concentration is improved through a repeated batch fermentation process, a repeated fed-batch fermentation process, a continuous fermentation process or a double-tank circulating continuous fermentation process; meanwhile, high acid production concentration, high production strength and high conversation rate are obtained. The conversation rate is 90% to a maximum, the production strength is 4.27g / L / h to a maximum, and the butyric acid yield is 98.7g / L to a maximum.

Description

technical field [0001] The invention relates to a method for fermenting and producing succinic acid by immobilizing Actinobacillus succinic acid with cotton fiber materials, belonging to the field of fermentation engineering. Background technique [0002] Succinic acid, also known as succinic acid, is an important C4 platform compound. At present, succinic acid is mainly produced by chemical synthesis using petroleum as raw material. The production of succinic acid by the fermentation of Actinobacillus succinic acid starts from renewable sugary raw materials and fixes CO through the TCA reduction pathway of microorganisms. 2 The production of succinic acid meets the development requirements of today's low-carbon economy and has good development prospects. However, the current production method is also restricted by high production costs. The production of succinic acid by microbial fermentation still has the problems of low bacterial biomass, high substrate concentration ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/46C12N11/10C12R1/01
Inventor 郑璞颜强
Owner SHANDONG XINGQIANG CHEM IND TECH RES INST CO LTD
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