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Technique for biotransforming CO2 into isopropyl alcohol by utilizing blue algae

A technology of isopropyl alcohol and cyanobacteria, applied in the field of recombinant cyanobacteria, can solve the problems of unable to achieve sustainable development of renewable energy, aggravate the shortage of fossil energy, and not solve the problem of developing bioenergy and food.

Inactive Publication Date: 2011-09-14
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method avoids the problem of exacerbating the shortage of fossil energy, Escherichia coli is a heterotrophic bacterium and still needs to use glucose to produce isopropanol. It does not solve the conflict between the development of bioenergy and food, and cannot achieve sustainable development of renewable energy. Therefore, the photoautotrophic bacteria cyanobacteria that can directly utilize solar energy are ideal hosts for the biosynthesis of isopropanol.

Method used

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  • Technique for biotransforming CO2 into isopropyl alcohol by utilizing blue algae
  • Technique for biotransforming CO2 into isopropyl alcohol by utilizing blue algae
  • Technique for biotransforming CO2 into isopropyl alcohol by utilizing blue algae

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Embodiment 1

[0070] Embodiment 1, application freshwater cyanobacteria produces isopropanol

[0071] Establishment of the isopropanol synthesis pathway in the freshwater cyanobacterium Synechocystis sp. 6803. Since there is acetyl-CoA acetyltransferase in Synechocystis sp., the genes encoding acetoacetyl-CoA transferase (ctfAB), acetoacetate decarboxylase (adc) and alcohol dehydrogenase (adh) are expressed in Synechocystis sp. ), the isopropanol synthesis pathway can be established.

[0072] 1. Construction of the recombinant plasmid pHR-ctfAB::adc::adh (Syneocystis homologous recombination integration expression vector)

[0073] 1. Acquisition of acetoacetyl-CoA transferase gene ctfAB and acetoacetate decarboxylase gene adc

[0074] ctfAB and adc are two adjacent genes on the Clostridium acetobutylicum genome. Therefore, the genomic DNA of Clostridium acetobutylicum was used as a template, and the primer pair composed of ctfABF and adcR was used for PCR amplification to obtain a PCR pr...

Embodiment 2

[0118]Embodiment 2, application freshwater cyanobacteria produces isopropanol and / or acetone

[0119] In the freshwater cyanobacterium Synechocystis 6803, the acetone pathway was established first, and the isopropanol pathway was established on the basis of acetone production. From figure 1 It can be seen from the synthesis route of isopropanol that isopropanol is derived from the dehydrogenation of acetone. Therefore, in this embodiment, it is carried out in two steps, that is, to obtain acetone-producing recombinant algae first, and then introduce alcohol dehydrogenase into the recombinant algae to obtain isopropanol-producing recombinant algae. Using the poly-β-hydroxybutyrate synthase gene (pha) on the Synechocystis genome as an integration platform, the genes encoding acetoacetyl-CoA transferase (ctfAB) and acetoacetate decarboxylase were expressed in Synechocystis sp. The gene (adc) was used to establish the acetone synthesis pathway; the gene encoding alcohol dehydrog...

Embodiment 3

[0170] Embodiment 3, application seawater cyanobacteria produces isopropanol (isopropanol synthesis pathway is established in seawater cyanobacterium Synechococcus 7002)

[0171] 1. Construction of recombinant plasmid pHR-thl::ctfAB::adc::adh (Synechococcus homologous recombination integration expression vector)

[0172] 1. Acquisition of the coding gene (thl) of acetyl-CoA acetyltransferase

[0173] Since there is no acetyl-CoA acetyltransferase in Synechococcus, it is necessary to express the enzyme in Synechococcus. The genomic DNA of Clostridium acetobutylicum was used as a template, and the primer pair composed of thlF and thlR was used for PCR amplification to obtain a PCR product (about 1100 bp), which was the thl gene.

[0174] thlF: 5'-ATGAAAGAAGTTGTAATAGC-3'

[0175] thlR: 5'-CTAGCACTTT TCTAGCAAT-3'

[0176] 2. The thl gene, the ctfAB-adc fusion gene of Example 1 and the adh gene of Example 1 are simultaneously used as templates, and the primer combination composed ...

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Abstract

The invention discloses a technique for biotransforming CO2 into isopropyl alcohol by utilizing blue algae, and provides the following six methods for preparing recombined blue algae: (1) leading a CoA transferase gene, an acetoacetate decarboxylase gene and an alcohol dehydrogenase gene into the blue algae; (2) leading the CoA transferase gene and the acetoacetate decarboxylase gene into the blue algae; (3) leading a coded gene of the alcohol dehydrogenase into the recombined blue algae in (2); (4) leading an acetyl-CoA acetyltransferase gene, an acetoacetyl-CoA transferase gene, the acetoacetate decarboxylase gene and the alcohol dehydrogenase gene into the blue algae; (5) leading the acetyl CoA acetyltransferase gene, the acetoacetyl-CoA transferase gene and the acetoacetate decarboxylase gene into the blue algae; and (6) leading the alcohol dehydrogenase gene into the recombined algae generating acetone in (5). The technique provided by the invention is one of ideal paths for realizing the continuous and healthy development of renewable and clean energy by utilizing the recombined algae to produce acetone and the isopropyl alcohol.

Description

technical field [0001] The invention belongs to the field of new biological energy and the field of environmental protection, and relates to a method of utilizing blue-green algae to convert CO 2 Technology for bioconversion to isopropanol, in particular to recombinant cyanobacteria capable of producing isopropanol. Background technique [0002] The energy problems and environmental problems facing the world have put forward the requirements for the development of renewable clean energy, and the food shortage requires the development of renewable clean energy to realize the goal of not competing with people for food, not for land, not for people and crops. freshwater. [0003] Blue-green algae, also known as cyanobacteria, is a prokaryotic organism that can perform oxygen-evolving photosynthesis. Cyanobacteria are widely distributed in fresh water, sea water and even sewage. They can reproduce rapidly using carbon dioxide and solar energy. They are ideal hosts for biosynth...

Claims

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Application Information

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IPC IPC(8): C12N1/13C12N15/63C12P7/04C12P7/28C12R1/89
Inventor 周杰张海峰张延平李寅
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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