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Method for producing hydrogen by bacterial fermentation

A technology for producing hydrogen and microorganisms, which is applied in the field of bacterial fermentation to produce hydrogen, and can solve problems such as little energy regulation

Inactive Publication Date: 2009-12-09
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the general research on improving hydrogen production capacity is the research on the metabolic pathways related to hydrogen production and the local regulation of enzymes, but there are few studies on the global energy regulation of the entire metabolism from the essence of hydrogen production.

Method used

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  • Method for producing hydrogen by bacterial fermentation

Examples

Experimental program
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Effect test

Embodiment 1

[0020] Embodiment 1, Enterobacter aerogenes uses sodium pyrophosphate as phosphorus source to produce hydrogen

[0021]1) Add sodium pyrophosphate to the basal medium (glucose 15.0g / L, tryptone 5.0g / L, ammonium sulfate 2.0g / L, magnesium sulfate 0.2g / L) so that the final concentration is 0.01mol / L, The pH was adjusted to 6.0, and the medium without sodium pyrophosphate was used as a control. Enter the activated Enterobacter aerogenes IAM1183 (Enterobacter aerogenes) in the medium, so that the initial content of Enterobacter aerogenes IAM1183 is 5×10 8 CFU / ml, cultured at 37°C, 170rpm for 15h.

[0022] 2) Add sodium pyrophosphate to the basal medium (glucose 15.0g / L, tryptone 5.0g / L, ammonium sulfate 2.0g / L, magnesium sulfate 0.2g / L) so that the final concentration is 0.10mol / L, The pH was adjusted to 6.0, and the medium without sodium pyrophosphate was used as a control. Enter the activated Enterobacter aerogenes IAM1183 (Enterobacter aerogenes) in the medium, so that the in...

Embodiment 2

[0031] Example 2. Recombinant Enterobacter aerogenes uses sodium pyrophosphate as phosphorus source to produce hydrogen

[0032] The pMCL plasmid is used as the expression vector of Enterobacter aerogenes IAM1183 (Enterobac ter aerogenes), and the pMCL plasmid construction process is as follows:

[0033] Use P1F and P1R primers (Table 1) to amplify the ampicillin resistance (Amp)-removed sequence on pMAL-c2X (purchased from NEB Company, No. N8076), and then digest with XhoI and KpnI to obtain product I;

[0034] PCR amplification conditions were: 94°C, 5min; 30 cycles, 94°C, 1min; 60°C, 3min; 72°C, 1min; final extension at 72°C for 5min, and finally 4°C storage.

[0035] Use primers P2F and P2R to amplify the chloramphenicol sequence (Cm) on pACYC184 (purchased from NEB Company, No. E4152S), and then clone it into the pMD-18T vector to obtain a recombinant vector. KpnI and XhoI digest the recombinant vector to obtain the product II;

[0036] PCR amplification conditions were...

Embodiment 3

[0049] Example 3, Enterobacter aerogenes produces hydrogen through aerobic-anaerobic conversion

[0050] 1) Add sodium dihydrogen phosphate to the basal medium (glucose 10.0g / L, tryptone 5.0g / L, ammonium sulfate 2.0g / L, magnesium sulfate 0.2g / L) to make the final concentration 0.01mol / L , the pH was adjusted to 6.0. Enter the activated Enterobacter aerogenes IAM1183 (Enterobacter aerogenes) in the culture medium, so that the initial content of Enterobacter aerogenes IAM1183 (Enterobacter aerogenes) is 5 × 10 8 CFU / ml, cultured at 37°C, 170rpm for 15h.

[0051]2) Add sodium dihydrogen phosphate to the basal medium (glucose 10.0g / L, tryptone 5.0g / L, ammonium sulfate 2.0g / L, magnesium sulfate 0.2g / L) to make the final concentration 0.10mol / L , the pH was adjusted to 6.0. Enter the activated Enterobacter aerogenes IAM1183 (Enterobacter aerogenes) in the culture medium, so that the initial content of Enterobacter aerogenes IAM1183 (Enterobacter aerogenes) is 5 × 10 8 CFU / ml, cu...

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Abstract

The invention discloses a method for producing hydrogen by bacterial fermentation, which is implemented by culturing hydrogen-producing microorganism in a basic culture medium added with phosphate radical ions to produce hydrogen under the conditions of (1) complete anaerobic culture and (2) culture with the concentration being 0.1-7.6% according to percent by volume, and then culture under complete anaerobic condition. The phosphate radical ions are H2PO4, HPO4, PO4, pyrophosphate radical or polyphosphate radical ions. The concentration of the phosphate radical ions is 0.01-0.15mol / L in respect to the basic culture medium. The hydrogen producing microorganism is enteroaerogen. The enteroaerogen is enteroaerogen or enteroaerogen expressing polyphosphate kinase. The hydrogen production method of the invention can improve the conversion yield of hydrogen and reduce production cost.

Description

technical field [0001] The invention relates to a method for producing hydrogen by bacterial fermentation. Background technique [0002] Biomass energy has always been an important energy source for human survival. It ranks fourth in the world's total energy consumption after coal, oil and natural gas, and occupies an important position in the entire energy system. In my country, biomass energy accounts for 20% of the total energy consumption. But for a long time, the proportion of biomass energy in my country's commercial energy structure is very small, and it is mainly used as primary energy in rural areas, accounting for about 70% of the total energy consumption in rural areas. As a relatively young research field, the biorefinery using biomass as raw material is mainly carried out in Europe; while the development of the industrial field is mainly in the United States. The U.S. Biomass Technology Advisory Council has launched a long-term plan and goal for bioenergy, bio...

Claims

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Application Information

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