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Escherichia coli YPC-SA-1 and application thereof

A YPC-SA-1, Escherichia coli technology, applied in the direction of bacteria, microorganisms, biochemical equipment and methods, etc., can solve the problems of unfavorable industrial production, long aerobic culture time, increased manpower, energy consumption, etc. Achieve great social significance and economic value, shorten the time of aerobic culture, and reduce the effect of culture time

Inactive Publication Date: 2016-05-18
SINOPEC YANGZI PETROCHEM +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in practical applications, the applicant found that during the aerobic growth stage of the bacterial strain, its growth was significantly slower in the culture medium prepared with industrial-grade raw materials than in the culture medium prepared with analytical-grade raw materials, showing a longer lag period, The total aerobic incubation time is also longer
In the process of industrial production, it is not feasible to use analytical-grade raw materials for production, while using industrial-grade raw materials requires longer cultivation time, which increases manpower, energy consumption, etc., and ultimately leads to an increase in product costs, which is not conducive to actual production. Industrial production

Method used

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  • Escherichia coli YPC-SA-1 and application thereof
  • Escherichia coli YPC-SA-1 and application thereof
  • Escherichia coli YPC-SA-1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] This example illustrates the method of performing the first step of plasma mutagenesis on the original Escherichia coli strain BER108.

[0039] The aerobic growth curve of the original starting strain BER108 in the synthetic medium prepared with analytical grade and industrial grade raw materials is as follows: figure 1 with figure 2 As shown, the growth lag period of strains using analytical-grade raw materials is 15h, and the growth is significantly inhibited, so the strains are selected by mutagenesis.

[0040] The method for carrying out the first step plasma mutagenesis of Escherichia coli starting strain is as follows:

[0041] The original strain of Escherichia coli BER108 (CN102643770A, deposit number CCTCCNO: M2012068) was activated in LB test tubes, cultured overnight at 37°C, 200r / min; freshly cultured cells were diluted to cell concentration OD 600 =1.0, drop it on a sterile slide, dry it with sterile wind; use helium as the discharge gas, 80~120W as the...

Embodiment 2

[0043] This example illustrates the method of screening excellent Escherichia coli.

[0044] Wherein, the medium formula used is as follows:

[0045] Solid synthetic medium plate: citric acid 3g / L, Na 2 HPO 4 12H 2 O4g / L, KH 2 PO 4 8g / L, (NH 4 ) 2 HPO 4 8g / L, (NH 4 ) 2 SO 4 0.75g / L,NH 4 Cl0.2g / L, MgSO 4 ·7H 2 O1g / L, CaCl 2 2H 2 O10.0mg / L, ZnSO 4 ·7H 2 O0.5mg / L, CuCl 2 2H 2 O0.25mg / L, MnSO 4 ·H 2 O2.5mg / L, CoCl 2 ·6H 2 O1.75mg / L, H 3 BO 3 0.12mg / L, Al 2 (SO4) 3 1.77mg / L, Na 2 MoO 4 2H 2 O0.5mg / L, iron citrate 16.1mg / L, vitamin B 1 20mg / L, biotin 2mg / L, agar 15g / L, glucose 10g / L.

[0046] Seed medium: peptone 10g / L, yeast powder 5g / L, NaCl 5g / L.

[0047] Synthetic medium: citric acid 3g / L, Na 2 HPO 4 12H 2 O4g / L, KH 2 PO 4 8g / L, (NH 4 ) 2 SO 4 0.75g / L, NH 4 Cl0.2g / L, MgSO 4 ·7H 2 O1g / L, CaCl 2 2H 2 O10.0mg / L, ZnSO 4 ·7H 2 O0.5mg / L, CuCl 2 2H 2 O0.25mg / L, MnSO 4 ·H 2 O2.5mg / L, CoCl 2 ·6H 2 O1.75mg / L, H 3 BO 3 0.12mg / L, Al 2 ...

Embodiment 3

[0059] This example illustrates the passage stability of the mutant strain YPC-SA-1.

[0060] On the synthetic medium plate, the mutant strain YPC-SA-1 was transferred and cultured several times, and the obtained strains were subjected to fermentation verification respectively. The experimental results are shown in Table 2:

[0061] Table 2 Passage stability analysis of mutant strain BEW308

[0062]

[0063] From the experimental results, it can be seen that after 8 consecutive subcultures, the aerobic growth of the mutant strain YPC-SA-1 is relatively stable, and it has good passage stability, which can be used as a production strain for further research and development.

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Abstract

The invention discloses escherichia coli which rapidly adapts to an industrial raw material medium for production of succinic acid and an application thereof. The classification term of the escherichia coli is Escherichia coli YPC-SA-1, and its preservation number is CCTCC NO:M 2014178. The invention also discloses an application of the above escherichia coli to industrial raw material cultivation growth. Under the aerobic condition, the strain can rapidly adapt to industrial raw material growth environment and grows by the utilization of nutrient composition, and lag phase is 7-8 h, which is almost the same under the condition of analytical raw material culture. In comparison with an original strain, lag phase of the strain in the industrial synthetic medium is shortened by 12 h, and aerobic culture time is shortened by 15 h. The mutant strain YPC-SA-1 has laid a foundation for industrial efficient fermentation for production of succinic acid.

Description

technical field [0001] The invention belongs to the technical field of industrial microorganism breeding and fermentation, and relates to Escherichia coli YPC-SA-1 and its application. 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 synthesize polybutylene succinate (PBS) Degradable polyester. [0003] In recent years, with the depletion of fossil resources and the increasingly serious environmental problems, the preparation of succinic acid by biologic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/46C12R1/19
Inventor 丁家海冯小华李维新沈品德黄树军黄兴东刘经纬方晓江陈韶辉
Owner SINOPEC YANGZI PETROCHEM
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