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Hydrogen producing engineering bacteria and application thereof

A technology for engineering bacteria and producing bacteria, applied in the directions of bacteria, microorganisms, biochemical equipment and methods, etc., to achieve the effect of increasing the production rate and the hydrogen production rate

Inactive Publication Date: 2009-12-30
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, hydrogen mainly comes from reforming conversion of fossil fuels (accounting for 96% of hydrogen sources) and hydrogen production by electrolysis of water (accounting for 4% of hydrogen sources), which obviously failed to get rid of the original fossil energy system

Method used

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  • Hydrogen producing engineering bacteria and application thereof
  • Hydrogen producing engineering bacteria and application thereof
  • Hydrogen producing engineering bacteria and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1. Regulation of intracellular NADH levels through NADH regeneration to increase bacterial hydrogen production rate

[0027] The NADH hydrogen production pathway is one of the important hydrogen production pathways of Enterobacter aerogenes. To increase the hydrogen production rate of cells by regulating the NADH metabolic pathway, it can be achieved by making the cells produce more NADH. Currently the most successful intracellular NADH regeneration system utilizes a system that relies on formic acid and NAD + NADH regeneration achieved by formate dehydrogenase (FDH1) (HCOO - +NAD + →NADH+CO 2 )(Riebel BR, Gibbs P R, Wellborn W B, et al. Cofactor regeneration of both NAD + fromNADH and NADP + from NADPH: NADH oxidase from Lactobacillus sanfranciscensis. Adv Synth Catal, 2003, 345).

[0028] Using the plasmid pMAL-c2X (purchased from NEB Company, No. N8076) as a template, and using P1F and P1R as primers (see Table 1 for specific primer sequences), PCR ampli...

Embodiment 2

[0040] Example 2. Regulation of NADH levels in cells through NADH regeneration to increase bacterial hydrogen production rate

[0041] The NADH hydrogen production pathway is an important hydrogen production pathway of Clostridium paraputrificum. To increase the hydrogen production rate of cells by regulating the NADH metabolic pathway, it can be achieved by making cells produce more NADH. Currently the most successful intracellular NADH regeneration system utilizes a system that relies on formic acid and NAD + NADH regeneration achieved by formate dehydrogenase (FDH1) (HCOO - +NAD + →NADH+CO 2 )(Riebel BR, Gibbs P R, Wellborn W B, et al. Cofactor regeneration of both NAD + fromNADH and NADP + from NADPH: NADH oxidase from Lactobacillus sanfranciscensis. Adv Synth Catal, 2003, 345).

[0042] Using the plasmid pMAL-c2X (purchased from NEB Company, No. N8076) as a template, and using P4F and P4R as primers (see Table 2 for specific primer sequences), PCR amplification was p...

Embodiment 3

[0055] Example 3. Regulating intracellular NADH levels by knocking out the lactic acid pathway that consumes the most NADH to increase bacterial hydrogen production rate

[0056] The NADH hydrogen production pathway is one of the important hydrogen production pathways of Enterobacter aerogenes. To increase the hydrogen production rate of cells by regulating the NADH metabolic pathway, more NADH can flow to the hydrogen production pathway by regulating the cell metabolism. In the generation and metabolism of cellular organic matter, the pathways that consume NADH include lactic acid production pathways, ethanol production pathways, succinic acid production pathways, and 2,3-butanediol production pathways. Among them, the lactic acid production pathway accounts for the largest proportion of cellular organic metabolites, and lactic acid is one of the main products of cellular anaerobic metabolism, consuming the most NADH. The hydrogen production rate can be increased by knocking ...

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Abstract

The invention discloses hydrogen producing engineering bacteria. The engineering bacteria are realized by over expressing a gene related to regeneration of nicotinamide adenine dinucleotide (NADH) in the hydrogen producing bacteria process and / or knocking out a metabolism path consuming the nicotinamide adenine dinucleotide in the hydrogen producing bacteria process. The metabolism path consuming the NADH can be knocked out, and a cell can generate more NADH through carrying out metabolic operation on an NADH hydrogen producing path of the hydrogen producing bacteria, so that the NADH as one of the most important complementary factors in the cell can be redistributed, and more metabolism flows can be promoted to enter the hydrogen producing path, and the hydrogen producing rate of the cell can be improved. Culture of the bacteria can greatly improve the hydrogen producing velocity with hydrogen yield in unit volume being 2.2 times of original bacteria, and improve the hydrogen producing rate for consuming glucose in unit mass by 86.8 percent, which basically approaches the theoretic yield of microbe produced hydrogen.

Description

technical field [0001] The invention relates to hydrogen-producing engineering bacteria and applications thereof. Background technique [0002] The fossil energy system established since the Industrial Revolution in the 18th century is facing two major challenges: the non-renewability of fossil fuels such as coal, oil, and natural gas; and environmental problems such as the greenhouse effect, acid rain, and dust pollution caused by the petroleum refining industry. These have greatly promoted the research and development of renewable energy industries such as wind energy, solar energy, water energy, biomass energy, geothermal energy and ocean energy. [0003] Biomass energy has always been an important source of energy for human survival, second only to coal, oil and natural gas, and ranks fourth in the world's total energy consumption, occupying an important position in the entire energy system. In my country, biomass energy accounts for 20% of the total energy consumption....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12P3/00C12R1/01C12R1/145
Inventor 邢新会卢元张翀来奇恒吴希
Owner TSINGHUA UNIV