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Recombinant plasmid and genetic engineering bacteria for producing succinic acid, production method and use thereof

A technology of genetically engineered bacteria and recombinant plasmids, applied in genetic engineering, plant genetic improvement, recombinant DNA technology, etc., can solve problems such as low yield, and achieve the effect of high succinic acid yield and increased growth rate

Inactive Publication Date: 2009-03-11
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] Although E. coli can also produce succinate itself, the yield is relatively low

Method used

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  • Recombinant plasmid and genetic engineering bacteria for producing succinic acid, production method and use thereof
  • Recombinant plasmid and genetic engineering bacteria for producing succinic acid, production method and use thereof
  • Recombinant plasmid and genetic engineering bacteria for producing succinic acid, production method and use thereof

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Embodiment Construction

[0049] The present invention will be further described below in combination with specific embodiments. However, these examples are only for illustrating the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following embodiments for which specific experimental conditions are not indicated are usually in accordance with conventional conditions, or in accordance with the conditions suggested by the manufacturer.

[0050] 1. The preparation of culture medium

[0051] The composition of LB medium is 5 g of yeast powder, 10 g of tryptone and 10 g of NaCl per liter of water. The solid medium is 1.5% agar powder added to the liquid base. In order to increase the selectivity of the medium, the concentration of ampicillin (Amp) used was 50 μg / ml.

[0052] 2. Extraction of total DNA from cyanobacterium Anabaena sp.7120

[0053] Cleavage with SDS-Protease K bacteria (cyanobacterium Anabaenasp.7120, purchased from the...

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Abstract

The invention discloses a recombinant plasmid for producing succinic acid and a succinic acid high-yield genetic engineering bacterium. The invention also discloses a method for preparing the recombinant plasmid and the genetic engineering bacterium. The invention simultaneously discloses application of the recombinant plasmid and the genetic engineering bacterium to the preparation of biological products for fermentation production of the succinic acid in the genetic engineering. The genetic engineering bacterium which is established by the recombinant plasmid has high stability, and improves the growth rate compared with the original strain BL21 (DE3); and the yield of the succinic acid is greatly improved under the condition of addition of revulsant with different concentrations, namely lactose or IPTG and two-stage fermentation of completely anaerobic or aerobic accumulation of biomasses and oxygen-free acid fermentation.

Description

technical field [0001] The invention belongs to the field of biological engineering. The invention relates to a recombinant plasmid for producing succinic acid and a genetic engineering bacterium with high succinic acid production. The present application also relates to the preparation method of the above-mentioned recombinant plasmid and genetically engineered bacteria. The present application also relates to the use of the above-mentioned recombinant plasmid and genetic engineering bacteria in the preparation of biological products for the production of succinic acid by genetic engineering fermentation. Background technique [0002] Energy and environmental protection are important issues facing the world today. As a biological material, succinic acid is one of the best petroleum substitutes. [0003] The scientific name of succinic acid is butanedioic acid, which is a linear saturated dicarboxylic acid and can be used as an intermediate chemical product to generate 1,...

Claims

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Application Information

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IPC IPC(8): C12N15/63C12N15/53C12N9/00C12N1/21C12P7/46
Inventor 邢建民王丹苏志国李望良李强
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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