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Method for producing medium-chain alkanes

A production method and paraffin technology, which is applied in the field of production of medium-carbon paraffins, and can solve the problem of producing very few paraffins

Inactive Publication Date: 2014-08-27
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Alkanes are mainly derived from oil and natural gas, and can also be produced through the metabolism of some species in nature, such as cuticle wax of plants, pheromones of insects, etc. very few

Method used

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  • Method for producing medium-chain alkanes
  • Method for producing medium-chain alkanes
  • Method for producing medium-chain alkanes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1 Cloning of acyl-CoA synthetase in Escherichia coli and construction of recombinant vector

[0039] 1.1 Cloning of acyl-CoA synthetase in Escherichia coli

[0040] The genome of Escherichia coli (Escherichia coli) K12 strain was extracted using the Bomide kit method, and the genome DNA of Escherichia coli was used as a template to design primers according to the GenBank sequence, and the corresponding enzyme cutting sites (NCOI and EcoRI) and protective bases were added, The acyl-CoA synthetase [acyl-CoA synthetase] gene encoded by fadD was amplified by PCR, and the acyl-CoA synthetase induced by the gene can metabolize fatty acids to generate ester acyl-CoA.

[0041] The primers used for PCR amplification of the acyl-CoA synthetase gene fadD are as follows:

[0042] Upstream primer: 5'-CATG CCATGG GCAAGAAGGTTTGGCTTAACC-3'

[0043] Downstream primer: 5'-CG GAATTC CGGCTCAGGCTTTATTGTCCACT-3'

[0044] The PCR reaction system contained 0.2 μL of genomic DNA, 0.25 μL...

Embodiment 2

[0051] 1 Cloning of acyl-CoA reductase in Acinetobacter and construction of recombinant vector

[0052] 1.1 Cloning of acyl-CoA reductase in Acinetobacter

[0053] The DNA of the acyl-CoA reductase [acyl-CoA reductase] gene in Acinetobacter baylyi was synthesized by the method of total gene synthesis. The gene-induced expression of acyl-CoA reductase can catalyze the reduction reaction of ester acyl-CoA accumulated in cells to generate C14 / C16 fatty aldehydes.

[0054] Using the acyl-CoA reductase gene DNA synthesized above as a template, primers were designed according to the GenBank sequence, and corresponding restriction sites (NCOⅠ and BamHI) and protective bases were added.

[0055] The primers used for PCR amplification of acyl-CoA reductase gene acr are as follows:

[0056] Upstream primer: 5'-GCAGCCCATGGCTAGCATGAACGCTAAAC-3'

[0057] Downstream primer: 5'-CCGCTGGATCCTTACCAGTGTTCACCTGG-3'

[0058] The PCR reaction system contained 0.2 μL of genomic DNA, 0.25 μL of u...

Embodiment 3

[0065] 1 Cloning of aldehyde decarbonylase from Nostoc algae and construction of recombinant vector

[0066] 1.1 Cloning of aldehyde decarbonylase from Nostoc algae

[0067] The aldehyde decarbonylase [aldehyde decarboxylase] gene DNA in Nostoc punctiforme was synthesized by the method of total gene synthesis. The aldehyde decarbonylase induced by this gene can catalyze the decarbonylation of aliphatic aldehydes to generate alkanes.

[0068] The gene is handed over to the company for synthesis, and after synthesis, it becomes the pUCE-dc plasmid and the strain containing the plasmid. Inoculate the glycerol tube of the strain into LB liquid medium for cultivation, and extract the plasmid with an omega kit, so that the plasmid containing the aldehyde decarbonylase gene dc can be cloned:

[0069] This gene sequence length is 699bp, utilizes 1% agarose gel electrophoresis to detect, and the obtained target DNA band is 699bp (such as Figure 5 shown).

[0070] 1.2 Construction ...

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Abstract

The invention belongs to the field of biochemical engineering, and particularly relates to a method for producing medium-chain alkanes. The alkanes are mainly derived from petroleum and natural gas and also can be produced from metabolism of species in the natural world, however the quantity of the alkanes of microorganisms is low. With the rapid development of modern molecular biology, particularly the development of synthetic biology, the expression of related enzymes in the synthetic process of the alkanes is regulated, so that microorganism cells can generate the alkanes only through simple metabolism, and an advanced technology for producing renewable energy is achieved. The method for producing medium-chain alkanes provides an excellent solution for the problems that traditional fossil fuel supply is insufficient, and novel biofuel can not completely replace the fossil fuel. Compared with other bacterial strains, escherichia coli has the advantages of being clear in genetic background, easy to improve, high in growing speed, suitable for high-density fermentation and the like, and is the ideal bacterial strain for synthetizing chemicals and fuel in a microbiological method. The method for producing the biological alkanes by the escherichia coli is a new way which does not occupy the space, does not depend on the weather, and can achieve sustainable production.

Description

technical field [0001] The invention belongs to the field of biochemical industry, in particular to a production method of medium-carbon paraffins. Background technique [0002] Alkanes are the main components of fossil energy, such as gasoline, diesel and aviation fuel, which are the separation products of petroleum, and are composed of alkanes with different carbon chain lengths. In recent years, with the depletion of fossil energy and the sharp increase in crude oil consumption, the shortage of fossil energy supply and how to achieve sustainable development have become issues that human beings must pay attention to. Therefore, research on how to use renewable energy to replace fossil fuels has gradually become a new hot topic. [0003] Currently the most widely used fossil energy substitutes mainly include fuel ethanol, fuel butanol and biodiesel. However, there are many problems in the application of these alternatives, such as alcohol gasoline is easy to form stratifi...

Claims

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Application Information

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IPC IPC(8): C12P5/00C12N15/70C12N1/21C12R1/19C12R1/01
CPCY02P20/59
Inventor 刘珞颜红王芳谭天伟邓利
Owner BEIJING UNIV OF CHEM TECH
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