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Method for reducing graphene oxide by utilizing microbial fermentation liquor

A microbial fermentation broth, graphene technology, applied in graphene, nano-carbon and other directions, can solve the problems of limited source of raw materials, complicated operation, high cost and so on

Pending Publication Date: 2020-09-04
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For example, researchers use environmentally friendly Vc as a reducing agent to synthesize reduced graphene oxide, but the high cost limits its development and application; or use extracts of natural environmental protection materials (such as: green tea extract) as reducing agents to synthesize reduced graphite oxide However, its disadvantages are limited source of raw materials and complex operation; Yong et al. developed a method for biological reduction of graphene oxide, the principle of which is to use electrons produced by microbial metabolism to reduce graphene oxide, the reaction conditions are mild, non-toxic and environmentally friendly, and Compared with the previous green reduction method, the operation is simple, the cost is low, and the source is wide, but the microorganisms are closely connected with the reduced graphene oxide, and the product separation is difficult

Method used

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  • Method for reducing graphene oxide by utilizing microbial fermentation liquor
  • Method for reducing graphene oxide by utilizing microbial fermentation liquor
  • Method for reducing graphene oxide by utilizing microbial fermentation liquor

Examples

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Effect test

Embodiment 1

[0028] Example 1: Construction of recombinant plasmids for heterologous expression of [FeFe] hydrogenase:

[0029] (1) Preparation of hydrogenase (HydA) and related mature enzymes (HydE, HydF, HydG):

[0030] First, with Clostridium acetobutylicum The NBRC 13948 genome (accession number: NC_003030.1) was used as a template, and hydrogenase (HydA) and its related mature enzymes (HydE, HydF, HydG) were amplified by high-fidelity PCR. The nucleotide sequences of hydrogenase (HydA) and its related mature enzymes (HydE, HydF, HydG) are shown in SEQ ID NO: 1-4, and the specific conditions of primer sequence, restriction site, annealing temperature and extension time are shown in the table 1 shows:

[0031] Table 1. PCR amplification primer sequences and reaction conditions

[0032]

[0033] (2) Construction of recombinant plasmid pETDuet-1-HydAE:

[0034] The pETDuet-1 plasmid (from Novagen) and the HydE gene fragment were simultaneously digested with Nde I and Kpn I sites,...

Embodiment 2

[0041] Example 2: Heterologous Expression of [FeFe] Hydrogenase E. coli Construction and Expression Confirmation of BL21(DE3)

[0042] (1) Construction method:

[0043] The recombinant plasmids pETDuet-1-HydAE and pCDFDuet-1-HydFG were simultaneously transferred into E. coli BL21 (DE3) competent (purchased from Sangon Bioengineering (Shanghai) Co., Ltd.), and spread evenly on LB plates containing 40 μg / mL streptomycin and 100 μg / mL ampicillin, after overnight incubation at 37 ℃ , to pick a single clone E. coli BL21(DE3)-HydAEFG culture.

[0044] (2) Heterologous expression of [FeFe] hydrogenase:

[0045] The recombinant strain was inoculated in 10 mL LB medium (containing 40 μg / mL streptomycin and 100 μg / mL ampicillin), and cultured overnight in a constant temperature shaker at 37 °C with a rotation speed of 200 rpm. Inoculate the overnight cultured bacterial solution into 200 mL LB liquid medium (add 100 mM 3-morpholine propanesulfonic acid (MOPS) solution, adjust ...

Embodiment 3

[0050] Embodiment 3: the method for reducing graphene oxide by microbial fermentation broth expressing hydrogenase heterologously

[0051] Get the fermented bacterium liquid anaerobic centrifugation of embodiment 2 and get the supernatant, under anaerobic condition, through 0.22 mu The m water system filter membrane is filtered and sterilized to obtain a sterile anaerobic fermentation broth. Add GO with a final concentration of 0.1-2 mg / mL and hydrogen with a final concentration of 0.1-50% to the fermentation broth, and carry out constant temperature shaking culture. The conditions are controlled at a temperature of 4-50 °C and a shaker speed of 100-250 rpm , The incubation time is 4-36 h, and GO is slowly reduced to rGO. Macro photo before reaction as image 3 As shown, both a and b show the light yellow transparent color of GO. The macroscopic photo after the reaction is as Figure 4 Shown: the fermented liquid in a changes from light yellow to yellowish brown without p...

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Abstract

The invention provides a method for reducing graphene oxide by utilizing microbial fermentation liquor, and belongs to the field of material synthesis. Genetically engineered Escherichia coli for heterologous expression of [FeFe] hydrogenase is constructed first, sterile fermentation liquor is obtained through treatment, and the sterile fermentation liquor is applied to reduction of graphene oxide. Therefore, biological green reduction of graphene oxide at normal temperature and normal pressure is realized.

Description

technical field [0001] The invention belongs to the field of material synthesis, and in particular relates to a method for reducing graphene oxide by using microbial fermentation broth. Background technique [0002] The carbon atoms in graphene are sp 2 The hybrid orbitals constitute a hexagonal honeycomb structure, and each carbon atom is perpendicular to the layer plane pz Orbitals can form polyatomic large π bonds throughout the entire layer (similar to benzene rings), have excellent electrical conductivity, optical and mechanical properties, and have important application prospects in materials science, environmental fields, biomedicine and drug delivery. [0003] The main preparation methods of graphene are: mechanical exfoliation method, epitaxy method, chemical vapor deposition method, silicon carbide epitaxy method and graphene oxide (GO) reduction method, etc. The principle of the graphene oxide reduction method is to oxidize graphite to obtain a dispersed graph...

Claims

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Application Information

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IPC IPC(8): C01B32/184
CPCC01B32/184
Inventor 王彦斋王兴强雍阳春
Owner JIANGSU UNIV
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