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Clostridium for synthesizing glutathion and construction method and use thereof

A technology of glutathione and clostridium, which is applied in the field of clostridium for the synthesis of glutathione and its construction and application, can solve the problem of strain production performance conversion rate, yield, production intensity and ability to withstand extreme environments that are difficult to meet industrial Production requirements and other issues, to achieve the effect of increasing butanol production intensity, improving downstream processing, and improving the ability to utilize starch

Inactive Publication Date: 2009-05-06
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the production performance of existing strains (including conversion rate, yield, production intensity and ability to withstand extreme environments) is still difficult to meet the requirements of industrial production

Method used

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  • Clostridium for synthesizing glutathion and construction method and use thereof
  • Clostridium for synthesizing glutathion and construction method and use thereof
  • Clostridium for synthesizing glutathion and construction method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1, construct the clostridium acetobutylicum of synthetic glutathione

[0036] The bacterial genome extraction kit was used to extract the chromosomal DNA of E.coli JM109 in the middle and late stages of logarithmic growth, and use this as a template to perform PCR amplification with the following primers:

[0037] gshB-F:5'-ttcaga ggatcc ATGATCAAGCTCGGCATCGTG-3' (the chemical line part is BamH I recognition site);

[0038] gshB-R:5'-aaggc gaattc TTACTGCTGCTGTAAACGTGCTTC-3' (the underlined part is the EcoR I recognition site).

[0039] The high-efficiency fidelity enzyme Primerstart of TaKaRa Company was used for PCR amplification, and the PCR amplification program was: 95°C for 3min; 98°C for 10s, 55°C for 15s, 72°C for 1min, 30 cycles; 72°C for 10min.

[0040] The obtained PCR product was purified using a PCR product recovery kit, and then the purified PCR product was double-enzymatically digested with BamH I and EcoR I from Fermetas, and after standing...

Embodiment 2

[0054] Embodiment 2, utilize Clostridium acetobutylicum (Clostridium acetobutylicum) SMB-1 (pITG-B) to produce butanol by fermentation

[0055] Clostridium acetobutylicum (Clostridium acetobutylicum) SMB-1 (pITG-B) was statically cultured in RCM medium at 37° C. to logarithmic phase, and used as fermentation seed solution 1.

[0056] The fermented seed liquid 1 was inoculated into a 7L fermenter equipped with 3L corn starch medium according to the volume percentage of 5% for fermentation. The fermentation temperature was 35°C, 37°C, and 39°C respectively, and the fermentation method was static fermentation; The time is 36 hours. Clostridium acetobutylicum (Clostridium acetobutylicum) SMB-1 (pIMP1) was used as a control. The fermented broth after fermentation was detected by liquid chromatography, and the results are shown in Table 1. Liquid chromatography detection condition is: sample pretreatment: 12000rpm centrifuges 1min, gets supernatant, filters with 0.22 μm filter mem...

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Abstract

The invention discloses a clostridium for synthesizing glutathione, a method for constructing the same and application thereof. The method for constructing the clostridium for synthesizing the glutathione is to introduce a DNA molecule containing an encoding gene of the glutathione for synthesizing related protein into the clostridium to construct a recombinant bacterium. The DNA molecule containing the encoding gene of the glutathione for synthesizing the related protein comprises a thiolase promoter and the encoding gene of the glutathione for synthesizing the related protein sequentially from the upstream to the downstream. The invention also discloses a method for producing butanol, which is to ferment and culture the recombinant bacterium constructed by the method for constructing the clostridium for synthesizing the glutathione to produce the butanol. The yield of the butanol produced through fermenting an acetone butanol clostridium for synthesizing the glutathione constructed by the invention is 12.8 grams per liter, and the highest yield thereof can reach 0.35 gram per liter per hour.

Description

technical field [0001] The invention relates to a clostridium for synthesizing glutathione, a construction method and application thereof. Background technique [0002] Butanol is an important chemical raw material, mainly used in the manufacture of plasticizers, solvents, extractants, etc. In recent years, it has been found that the calorific value and octane number of butanol are equivalent to those of gasoline, and it can be mixed with gasoline in any ratio, and can be transported by existing oil pipelines. A promising new biofuel. The production of butanol by biorefinery has a long history, and a lot of research has been carried out from the breeding of excellent strains to the improvement of production technology and extraction technology. However, the production performance of the existing strains (including conversion rate, yield, production intensity and ability to withstand extreme environments) is still difficult to meet the requirements of industrial production....

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/54C12P7/16C12R1/145
Inventor 朱林江张延平李寅
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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