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Method for producing pimelic acid by fermenting enterobacter hormaechei

A technology of Enterobacter horii and pimelic acid is applied in the field of Enterobacter horii fermenting and producing pimelic acid to achieve the effect of reducing the cost of industrialization

Active Publication Date: 2022-07-08
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the current problems, the present invention provides a screening method for high-yield pimelic acid strains and its fermentation optimization method

Method used

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  • Method for producing pimelic acid by fermenting enterobacter hormaechei
  • Method for producing pimelic acid by fermenting enterobacter hormaechei
  • Method for producing pimelic acid by fermenting enterobacter hormaechei

Examples

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Effect test

Embodiment 1

[0037] Example 1: Screening of pimelic acid-producing strains

[0038] After streaking the strains preserved in the laboratory, pick a single colony to prepare a secondary seed solution: the single colony of the inoculated strain is placed in a 250 mL Erlenmeyer flask containing 50 mL of LB medium, and cultured at 37°C for 12 hours at 250 r / min to prepare primary seeds. Continue to transfer the first-class seed solution to a conical flask at 2% of the inoculum, and cultivate at 37°C at 250 r / min for 12 h to prepare the second-class seed solution.

[0039] A 48-well plate was used for primary screening, fermentation was performed at 30°C, 250 r / min for 72 h, and pimelic acid in the fermentation broth was detected by HPLC. Then, the strains obtained from the primary screening were re-screened in a shaker flask, and the prepared secondary seed liquid was inoculated into a 250mL conical flask containing 50mL SOB medium with 2% inoculum, and fermented at 30°C and 250r / min for 72h. ...

Embodiment 2

[0041] Example 2: Optimization of media composition

[0042] Preparation of secondary seed solution: inoculate a single colony of strain Bpa-3 in a 250 mL conical flask containing 50 mL of LB medium, and cultivate at 37°C for 12 hours at 250 r / min to prepare primary seed solution; inoculate the primary seed solution with 2% Continue to transfer the amount to the Erlenmeyer flask, cultivate at 37°C, 250r / min for 12h to prepare secondary seed solution (OD 600 =4±0.5).

[0043] (1) Optimization of carbon source species: 4g / L sucrose and 4g / L glycerol were used to replace glucose in SOB medium, respectively, and the secondary seed solution of strain Bpa-3 was treated with an inoculum of 2% (v / v). It was inoculated into the medium, fermented at 30°C, 250r / min for 72h, sampling every 12h, and the fermentation broth was analyzed by HPLC. The result is as figure 2 As shown, after 72 h of fermentation, when the carbon source was glycerol, the yield of pimelic acid was 122.5 mg / L as...

Embodiment 3

[0047] Example 3: Optimization of inoculum size

[0048] The secondary seed liquid of strain Bpa-3 was transferred with the inoculum of 1%, 2%, 4%, 6%, 8% and 10% (v / v) respectively, and the optimal composition medium was used for shake flask fermentation , 30 ℃, 250r / min fermentation 72h, sampling every 12h, using HPLC to analyze the fermentation broth. The result is as Figure 5 As shown, when the inoculation amount is 4%, the yield of pimelic acid can reach a maximum of 149.2 mg / L, and when the inoculation amount is 1%, 2%, 6%, 8%, and 10%, the yield of pimelic acid is respectively were 113.4 mg / L, 123.5 mg / L, 140.3 mg / L, 140.8 mg / L, and 124.0 mg / L.

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Abstract

The invention discloses a method for producing pimelic acid through fermentation of enterobacter hormaechei, and belongs to the technical field of microbial fermentation. The enterobacter hormaechei capable of producing pimelic acid is screened from preserved strains, the strain is a natural strain, antibiotics and inducers do not need to be added in the fermentation process, the industrial production cost is reduced, and meanwhile environment protection is facilitated. When the enterobacter hormaechei is fermented for 72 hours under a shake flask condition, the yield of pimelic acid is 115.4 mg / L; by optimizing the fermentation processes such as the carbon source type, the nitrogen source type, the carbon and nitrogen source combination, the inoculum size, the type and concentration of additional inorganic salt and the like, the yield of pimelic acid is further increased and reaches 217.3 mg / L. Meanwhile, a new host is provided for biological synthesis of pimelic acid, and a biological foundation is also laid for subsequent research.

Description

technical field [0001] The invention relates to a method for fermenting and producing pimelic acid by Enterobacter hallii, and belongs to the technical field of microbial fermentation. Background technique [0002] Pimelic acid, also known as tartaric acid, is an important seven-carbon dicarboxylic acid, mainly used as a precursor for biotin synthesis, in addition to the preparation of polyamides, surfactants and pesticides . At present, pimelic acid is mainly synthesized industrially by the cracking of salicylic acid, but this method has disadvantages such as complicated process and low yield. [0003] With the continuous development of modern biotechnology, biosynthesis of pimelic acid has gained more and more attention, and the existence of trace amounts of pimelic acid was first discovered in the culture process of some microorganisms. Subsequently, the biosynthesis of pimelic acid was studied by means of metabolic engineering and synthetic biology. By overexpressing ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/44C12R1/01
CPCC12P7/44Y02A50/30
Inventor 邓禹李国辉鲍青青支睿毛银赵运英周胜虎
Owner JIANGNAN UNIV
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