Biological hydrogen production method and special strain thereof

A technology of recombinant bacteria and host bacteria, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems that cannot reach the level of practical application, achieve improved hydrogen production capacity, broad application prospects, and energy saving Effect

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

[0006] At present, research at home and abroad is mainly based on strain screening, domestication, breeding and optimization of various fermentation process parameters, which have not yet reached the level of practical application.

Method used

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  • Biological hydrogen production method and special strain thereof
  • Biological hydrogen production method and special strain thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1. Using Enterobacter aerogenes as a host, the gene encoding the regulatory factor FNR was transferred to improve its hydrogen production capacity

[0068] 1. Construction of recombinant bacteria

[0069] The expression plasmid pMCL-fnr was electrotransfected into Enterobacter aerogenes IAM1183 to obtain recombinant Enterobacter aerogenes / pMCL-fnr containing gene fnr.

[0070] 2. Fermentation of recombinant Enterobacter aerogenes / pMCL-fnr to produce hydrogen

[0071] The medium used consists of: every liter of medium contains glucose 15g, peptone 5g, (NH 4 ) 2 SO 4 2g, KH 2 PO 4 14g,K 2 HPO 4 3H 2 O 6g, MgSO 4 0.2 g, and the pH of the culture medium was 6.0.

[0072] Insert the activated recombinant Enterobacter aerogenes / pMCL-fnr into the medium so that its initial content in the medium is 5×10 8 CFU / ml, cultured at 37°C and 170rpm for 15h, and the recombinant plasmid was induced with IPTG (0.5mM). During the cultivation process, anaerobic condition...

Embodiment 2

[0080] Example 2, using Enterobacter aerogenes as a host, transferring the coding gene of the regulatory factor NarP to improve its hydrogen production capacity

[0081] 1. Construction of recombinant bacteria

[0082] The expression plasmid pMCL-narp was electrically introduced into Enterobacter aerogenes (Enterobacter aerogenes) IAM1183 to obtain recombinant Enterobacter aerogenes / pMCL-narp containing gene narp.

[0083] 2. Fermentation of recombinant Enterobacter aerogenes / pMCL-narp for hydrogen production

[0084] The cultivation method of the recombinant bacteria, the culture medium used, the detection method of the hydrogen production, the calculation method of the hydrogen production rate, etc. are all as described in Example 1.

[0085] At the same time, the recombinant Enterobacter aerogenes / pMCL transformed into the empty plasmid pMCL was used as a control.

[0086] Experiments were repeated three times and results are presented as mean ± standard deviation. The h...

Embodiment 3

[0088] Embodiment 3, using Clostridium putrefaction-like as host, transfer the coding gene of regulatory factor FNR to improve its hydrogen production capacity

[0089] 1. Construction of recombinant bacteria

[0090] The expression plasmid pMKL-fnr was electrotransfected into Clostridium paraputrificum (Clostridium paraputrificum) ATCC17796 to obtain recombinant Clostridium paraputrificum / pMKL-fnr containing gene fnr.

[0091] 2. Fermentation bacteria produce hydrogen

[0092] The composition of the medium used is: glucose 15.0g / L, tryptone 2.0g / L, yeast extract 4.5g / L, sodium carbonate 4.0g / L, cysteine ​​hydrochloride 3.0g / L; to 6.0.

[0093] Insert the activated recombinant Clostridium putrefaction-like / pMKL-fnr into the culture medium so that its concentration in the culture medium is 5×10 8 CFU / ml, the recombinant plasmid was induced with IPTG (0.5mM), and cultured at 37°C and 170rpm for 15h. During the cultivation process, anaerobic conditions were adopted, and oxyge...

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Abstract

The invention discloses a biohydrogen production method and a special strain thereof. The special strain is a recombination strain which is obtained from introducing genes inciting anaerobic fermentation of the host strain into the hydrogen production host strain, positively correlating with the host strain anaerobic metabolism and containing the introduced genes. Proved by tests, the hydrogen production capability of the recombination strain is obviously improved compared with the hydrogen production capability of a contrast strain; therefore, the recombination strain and the biohydrogen production method of the invention have wide application prospect in biohydrogen production field and great significance for saving energy.

Description

technical field [0001] The invention relates to a biological hydrogen production method and special bacteria. 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, ...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C12N1/21C12P3/00C12R1/01C12R1/145C12R1/185
Inventor邢新会卢元张翀来奇恒
OwnerTSINGHUA UNIV