Escherichia coli LL016 and application thereof

A technology of Escherichia coli and succinic acid, applied in the direction of bacteria, microorganisms, biochemical equipment and methods, etc.

Inactive Publication Date: 2013-02-20
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, under pure anaerobic conditions, there are few reports on the growth and accumulation of succinic acid in Escherichia coli using synthetic media.

Method used

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  • Escherichia coli LL016 and application thereof
  • Escherichia coli LL016 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] This example illustrates the method of performing the first step of plasma mutagenesis on the original strain of Escherichia coli.

[0038] The method for carrying out the first step plasma mutagenesis of Escherichia coli original strain is as follows:

[0039] Activate the original strain of Escherichia coli BA016 in an LB test tube, culture overnight at 37°C, 200 r / min; take freshly cultured cells and dilute to the cell concentration OD 600 =1.0, drop on the sterile slide, and dry with sterile wind; use helium as the discharge gas, 80-120 W as the radio frequency power, 10-30 SLM as the gas flow rate, and 10-30 s as the radiation According to the time, plasma mutagenesis was performed on the strain; after mutagenesis, the bacterial film on the slide was eluted with 1 mL of sterile saline, spread on a synthetic medium plate, and cultivated in an anaerobic box. The experimental results are attached figure 1 , where the abscissa is the irradiation time, and the ordina...

Embodiment 2

[0041] This example illustrates the method for screening excellent Escherichia coli.

[0042] Wherein, the medium formula used is as follows:

[0043] (1) Solid synthetic medium plate: citric acid 3 g L -1 , Na 2 HPO 4 12H 2 O 4 g L -1 , KH 2 PO 4 8 g L -1 , (NH 4 ) 2 HPO 4 8 g L -1 , NH 4 Cl 0.2 g L -1 , (NH 4 ) 2 SO 4 0.75 g·L -1 , MgSO 4 ·7H 2 O 1 g L -1 , CaCl 2 2H 2 O 10.0 mg·L -1 , ZnSO 4 ·7H 2 O 0.5 mg L-1 , CuCl 2 2H 2 O 0.25 mg L -1 , MnSO 4 ·H 2 O 2.5 mg L -1 , CoCl 2 ·6H 2 O 1.75 mg L -1 , H 3 BO 3 0.12 mg·L -1 , Al 2 (SO 4 ) 3 1.77 mg·L -1 , Na 2 MoO 4 2H 2 O 0.5 mg L -1 , ferric citrate 16.1 mg·L -1 , Vitamin B 1 20 mg·L -1 , biotin 2 mg·L -1 , agar 20 g / L, glucose 10 g / L.

[0044] (2) Seed medium: peptone 10 g / L, yeast powder 5 g / L, NaCl 5 g / L.

[0045] (3) Shake flask fermentation medium: citric acid 3 g L -1 , Na 2 HPO 4 12H 2 O 4 g L -1 , KH 2 PO 4 8 g L -1 , (NH 4 ) 2 HPO 4 8 g L -1 , NH 4...

Embodiment 3

[0057] This example illustrates the passage stability of the mutant strain LL016

[0058] On the synthetic medium plate, the mutant strain LL016 was transferred and cultured several times, and the obtained strains were subjected to fermentation verification respectively. The experimental results are shown in Table 2:

[0059] Table 2 Passage stability analysis of mutant strain LL016

[0060]

[0061] It can be seen from the experimental results that after 8 consecutive subcultures, the succinic acid production and succinic acid conversion rate of the mutant strain LL016 are relatively stable, with good passage stability, and can be used as a production strain for further research and development.

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PUM

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Abstract

The invention discloses Escherichia coli LL016 with a preservation number of CCTCC (China Center For Type Culture Collection) NO:M2012429. The invention also discloses application of the Escherichia coli LL016 to production of butanedioic acid by using a synthetic medium through pure anaerobic fermentation, and an application method of the Escherichia coli LL016. The Escherichia coli grows by using an inorganic nitrogen source and glucose in an anaerobic condition, greatly accumulates butanedioic acid, and has the advantages of higher growth rate, higher acid yield and great social significance and high economic value.

Description

technical field [0001] The invention relates to a strain of Escherichia coli LL016 and its application, in particular to a strain of Escherichia coli LL016 capable of producing succinic acid by pure anaerobic fermentation of a synthetic medium and its application, belonging to industrial microbial breeding and its fermentation technology field. Background technique [0002] Succinic acid, also known as succinic acid, is a common natural organic acid widely found in the human body, animals, plants and microorganisms. As an intermediate product of the TCA cycle and one of the terminal reduction products of anaerobic metabolism, succinic acid plays a very important role in the process of biological metabolism. Research results in recent years have shown that succinic acid can also be used as a C4 platform compound to synthesize bulk chemicals such as 1,4-butanediol, tetrahydrofuran, and γ-butyrolactone, as well as polybutylene succinate (PBS) biodegradable compounds. Degrades...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/18C12R1/19
Inventor 姜岷梁丽亚刘嵘明万青陈旭任心怡马江锋陈可泉韦萍欧阳平凯
Owner NANJING UNIV OF TECH
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