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Enterobacteria recombinant strain and use thereof

A technology of Enterobacteriaceae and recombinant bacteria, applied in the direction of recombinant DNA technology, bacteria, microorganisms, etc., can solve the problems of aggravating the greenhouse effect, reducing the conversion rate of substrates into products, etc., achieving broad application prospects, reducing CO2 production, and improving conversion rate effect

Inactive Publication Date: 2011-07-20
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

CO 2 The generation of the substrate not only caused the reduction of the conversion rate of the substrate into the product, but also exacerbated the already increasingly serious greenhouse effect.

Method used

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  • Enterobacteria recombinant strain and use thereof
  • Enterobacteria recombinant strain and use thereof
  • Enterobacteria recombinant strain and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1, the construction of the Klebsiella oxytogenes recombinant bacterium that pyruvate formate lyase gene is inactivated

[0031] 1. Cloning of pyruvate formate lyase gene pfl and its upstream and downstream sequences

[0032] Primers p1 and p2 were designed according to the partial genome sequence of Klebsiella oxytoca (Genome Project ID: 12627) and the sequence of pyruvate formate lyase gene pfl (EU276019), and the pyruvate formate lyase gene pfl and its Upstream and downstream sequences, primer sequences are as follows:

[0033] p1 (upstream primer): 5'-TCAGGTACCTTGACAGCGATTTGATGGCT-3' (sequence 1 in the sequence listing),

[0034] p2 (downstream primer): 5'-AGTGAGCTCGAAGCGTTCATAAAGTGGCG-3' (sequence 2 in the sequence listing); the primer was synthesized by Beijing Saibaisheng Company.

[0035] Wild-type Klebsiella oxytoca M5al (Ohta K, Beall DS, Mejia JP, et al. Metabolicengineering of Klebsiella oxytoca M5A1 for ethanol product ion from xylose and glucos...

Embodiment 2

[0045] Embodiment 2, the glucose anaerobic fermentation of the Klebsiella oxytogenes recombinant bacterium that pyruvate formate lyase gene is inactivated

[0046] Klebsiella oxytoca uses glucose as a substrate to ferment and can produce lactic acid, succinic acid, 2,3-butanediol and by-product CO 2 、H 2 . In this experiment, the recombinant bacteria constructed in Example 1 were used for anaerobic fermentation, and the wild-type strain (ie, the starting bacteria) was used as a control to compare the results of its products and by-products. The specific method is as follows:

[0047] 1. Seed cultivation

[0048] Pick the single bacterium colony of wild-type Klebsiella oxytocia (starting bacterium) or the recombinant bacterium of Klebsiella oxytogen obtained in embodiment 1 from the plate, insert into 500 milliliters triangles that 100 milliliters of LB culture medium are housed In the bottle, cultivate at 37°C and 180r / min for 12 hours to obtain a seed culture solution.

...

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Abstract

The invention discloses a recombination strain for an enterobacterium and application of the recombination strain for the enterobacterium. The recombination strain provided by the invention is a recombination strain obtained after deactivation of a pyruvate formate-lyase gene in the enterobacterium. As shown by experiments, the amount of by-product CO2 produced during the fermentation process of the recombination strain is obviously reduced and the amount of target products is obviously improved compared with the control. For example, the amount of lactic acid, the amount of succinic acid and the amount of 2, 3-butanediol produced by fermentation of klebsiella oxytoca are respectively improved by 30 percent, 9 percent and 23 percent compared with the control, and the amount of the CO2 produced is only 8.04 percent compared with the control. The recombination strain helps to release the pressure of the greenhouse effect, has important function in the actual industrial production,not only can greatly reduce the yield of the CO2, but also can improve the conversion rate of converting substrates into products, and has wide application prospect.

Description

technical field [0001] The invention relates to a recombinant bacterium and its application, in particular to a recombinant bacterium of Enterobacter and its application. Background technique [0002] In the fermentation industry, when the substrate generates the target product under the action of microorganisms or enzymes, by-products are often produced, CO 2 It is one of the most common by-products in the fermentation industry. During the anaerobic fermentation of sugars by strains of Enterobacteriaceae, sugars are converted into phosphoenolpyruvate through a series of reactions, a part of phosphoenolpyruvate is converted into succinic acid, and the other part is converted into pyruvic acid, pyruvic acid There are three outlets: 1. converted into lactic acid under the catalysis of lactate dehydrogenase; ) catalyzed by α-acetolactate and 3-hydroxybutanone into 2,3-butanediol in turn; 3. decomposed into acetyl CoA and formic acid under the catalysis of pyruvate formate lya...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12N15/09C12P7/56C12P7/46C12P7/18C12R1/22C12R1/185C12R1/01C12R1/42C12R1/425
Inventor 杨光曹竹安
Owner TSINGHUA UNIV
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