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Method for producing metal nanoparticles and metallic sulfide nanoparticles using recombinant microorganisms

A technology of metal nanoparticles and recombinant microorganisms, applied in biochemical equipment and methods, preparation of sulfide/polysulfide, metallothionein, etc., can solve the problems of unfavorable production cost and environmental protection

Active Publication Date: 2018-09-28
KOREA ADVANCED INST OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the above-mentioned methods are disadvantageous in that they require a high reaction temperature of 300°C or higher, are not environmentally friendly due to the use of an organic solvent such as toluene, and require a subsequent heat treatment process, which is disadvantageous in terms of production cost

Method used

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  • Method for producing metal nanoparticles and metallic sulfide nanoparticles using recombinant microorganisms
  • Method for producing metal nanoparticles and metallic sulfide nanoparticles using recombinant microorganisms
  • Method for producing metal nanoparticles and metallic sulfide nanoparticles using recombinant microorganisms

Examples

Experimental program
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Effect test

Embodiment 1

[0084] Embodiment 1: the expression load of co-expressing two kinds of heavy metal adsorption proteins (metallothionein and phytochelatin synthase) body construction

[0085] 1-1: Construction of metallothionein expression vector

[0086] Using the genomic DNA of Pseudomonas putida (Pseudomonas putida) KT2440 as a template, the primers of SEQ ID NO: 3 and 4 were used to carry out polymerase chain reaction (PCR), thereby constructing the nucleotide sequence encoding metallothionein (MT) The metallothionein (MT) gene fragment.

[0087] SEQ ID NO:3:5'-ATAGAATTCATGAACGATCACCACCACCACAAC-3'

[0088] SEQ ID NO:4:5'-TATCTGCAGTTAGGGCGAGATCGGATCACTC-3'

[0089] Next, the constructed metallothionein fragment was subjected to agarose gel electrophoresis to separate the 243-bp metallothionein gene fragment, which was then digested with two restriction enzymes (EcoRI and PstI). Simultaneously, the inducible pTac15K plasmid containing the tac promoter was digested with two restricti...

Embodiment 2

[0102] Example 2: Synthesis and Characterization of Elemental Metal Nanoparticles Using Recombinant Microorganisms

[0103] 2-1: Synthesis of elemental metal nanoparticles using recombinant microorganisms

[0104] One hour after induction of expression of metallothionein and phytochelatin synthase in the recombinant microorganisms produced in Example 1 above by IPTG, barium (Ba, (CH 3 COO) 2 Ba, 0.5mM), zirconium (Zr, K 2 ZrF 6 , 0.5mM), molybdenum (Mo, Na 2 MoO 4 2H 2 O, 0.5mM), indium (In, InCl 2 4H 2 O, 0.5mM), tin (Sn, SnCl 2 4H 2 O, 0.5mM), lanthanum (La, La(NO 3 ) 3 6H 2 O, 0.5mM), cerium (Ce, Ce(NO 3 ) 3 6H 2 O, 0.5mM) or praseodymium (Pr, Pr(NO 3 ) 3 6H 2 (2, 0.5mM) has been added in the LB liquid culture medium (tryptone 10g / L, yeast extract 5g / L, NaCl 5g / L, pH 6.5) that has cultivated recombinant E. Incubate for 12 hours. The culture was centrifuged at 3500 rpm and 4°C for 15 minutes, after which time the supernatant was discarded and the E. c...

Embodiment 3

[0109] Example 3: Synthesis and Characterization of Metal Alloy Nanoparticles Using Recombinant Microorganisms

[0110] 3-1: Synthesis of metal alloy nanoparticles using recombinant microorganisms

[0111] One hour after induction of expression of metallothionein and phytochelatin synthase by IPTG in the recombinant microorganism produced in Example 1 above, i) cobalt and iron (Co, CoCl 2 ,0.5mM, Fe, Fe(NO) 3 ·6H2 O, 0.5mM), ii) nickel and iron (Ni, NiCl 2 ·6H 2 O, 0.5mM, Fe, Fe(NO) 3 ·6H 2 O, 0.5mM), iii) zinc and manganese (Zn, ZnSO 4 ·7H 2 O, 0.5mM, MnCl 2 4H 2 O), 0.5mM, or iv) zinc and iron (Zn, ZnSO 4 ·7H 2 O, 0.5mM, Fe, Fe(NO) 3 ·6H 2 (2, 0.5mM) ions are added in the LB liquid culture medium (tryptone 10g / L, yeast extract 5g / L, NaCl5g / L, pH 6.5) that has cultivated recombinant escherichia coli bacterial strain, after this this recombinant microorganism is additionally Incubate for 12 hours. The culture was centrifuged at 3500 rpm and 4°C for 15 minutes...

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Abstract

The present invention relates to a method for producing metal nanoparticles and metal sulfide nanoparticles using recombinant microorganisms in which phytochelatin synthase and metallothionein are simultaneously expressed as heavy metal adsorption proteins, and a use of metal nanoparticles or metallic sulfide nanoparticles synthesized by the method. According to the present invention, by providinga method for synthesizing metal in the form of nanoparticles, in which the metal was difficult to synthesize by a conventional biological method, it is possible to synthesize metal nanoparticles in an eco-friendly and economical manner, and it is also possible to synthesize metal sulfide nanoparticles using metal and sulfur, and furthermore, the metal nanoparticles conventionally produced by a chemical or biological method significantly increase in yield by using the production method of the present invention.

Description

technical field [0001] The present invention relates to methods of producing metal nanoparticles and metal sulfide nanoparticles using recombinant microorganisms, and more particularly to producing metal nanoparticles and metal sulfide nanoparticles using recombinant microorganisms co-expressing metallothionein and phytochelatin synthase The method of the present invention, the metallothionein and the phytochelate synthase are heavy metal adsorption proteins. Background technique [0002] As interest in nanotechnology has increased, various studies have been actively conducted in various fields including physics, chemistry, materials science, electricity, and electronics to develop new materials at the nanoscale. Specifically, materials containing metallic nanostructures can be applied in the fabrication of high-efficiency electronics, optics, optoelectronics, electronic devices, bioactive molecular detection devices, and catalysts, which are difficult to achieve using bulk ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P3/00C12N9/10C12N15/52C12N15/63C01B17/20C01G5/00C01G15/00C01G19/00C01G45/00
CPCC01G19/00C01G45/00C01B19/002C01G5/00C01G15/00C01P2002/72C01P2002/82C01P2004/04C01P2004/64C01P2006/40C01P2006/42C07K14/825C12N1/20C01G45/1235C01G49/0063C01G51/40C01G53/40C01G1/12C12N9/0051C12P3/00C12N9/104C12Y203/02015C01B17/20C12N15/52C12N15/63
Inventor 李相烨崔琟真
Owner KOREA ADVANCED INST OF SCI & TECH
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