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A surface-displayed metal-binding protein gene and its application in recycling platinum and palladium metal

A technology for metal-binding proteins and surface display, which is applied to the surface display of metal-binding protein genes and its application in the recovery of platinum and palladium metals, can solve the problems of poor adsorption and poor selectivity of platinum group metals, and achieve low cost and low cost. , no secondary pollution effect

Active Publication Date: 2020-11-06
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a surface-displayed metal-binding protein gene and its application in the recovery of platinum and palladium metals, which solves the problems of poor selectivity and poor adsorption of microorganisms to platinum group metals

Method used

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  • A surface-displayed metal-binding protein gene and its application in recycling platinum and palladium metal
  • A surface-displayed metal-binding protein gene and its application in recycling platinum and palladium metal
  • A surface-displayed metal-binding protein gene and its application in recycling platinum and palladium metal

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] (1) Synthesis of protein and metal binding protein genes

[0026] The gene sequences of the anchor protein (InaKN) and the metal binding protein (EC20) were sent to a biological company for synthesis and optimization, making them suitable for expression in Escherichia coli. The synthetic gene sequence was connected to the pET28a(+) plasmid, in which the InaKN sequence was inserted between the Nco I and Hind III restriction sites, and the EC20 sequence was inserted between the NotI and XhoI restriction sites to obtain the recombinant plasmid pET28a(+)- InaKN-EC20, which contains the nucleotide sequence shown in SEQ ID NO.3.

[0027] (2) Transform E.coli BL21 Competent Cells with Recombinant Plasmid

[0028] ①Put 100 μL of E.coli BL 21 competent cells into a centrifuge tube, then put the centrifuge tube in an ice bath, then add 10 μL of recombinant plasmid pET28a(+)-InaKN-EC20 to the centrifuge tube, flick and mix well, Put it in an ice bath and let it stand for 30min. ...

Embodiment 2

[0046] In this embodiment, the palladium (Pd) metal solution used has a concentration of 0.5, 2.5, 5, 25, 50, 100, and 200 mg / L, and the adsorption pH is 3.0. The specific operation steps of the experiment are as follows (combined with figure 1 ):

[0047] (1) The preparation of microbial adsorbent, concrete steps are:

[0048] The E.coli BL21 original bacteria were inoculated into sterilized LB medium, cultured at 30°C and 170rpm for 24h, and then centrifuged at 10000rpm for 10min to collect the bacteria. The E.coli BL21 engineering bacteria prepared in Example 1 were inoculated into the HB-PET self-inducing medium, cultured at 30° C. and 170 rpm for 24 hours, and then centrifuged at 10,000 rpm for 10 minutes to collect the bacteria. Among them, the self-inducing medium was purchased from Qingdao Hi-Tech Park Haibo Biotechnology Co., Ltd., and 1 L of the medium contained 27.5 g of culture powder and 5 g of glycerol.

[0049] (2) The preparation of Pd (II) stock solution, c...

Embodiment 3

[0055] In this embodiment, the platinum (Pt) metal solution used has a concentration of 1, 5, 10, 50, 100, and 200 mg / L, and the adsorption pH is 3.0. The specific operation steps of the experiment are as follows:

[0056] (1) The preparation of microbial adsorbent, concrete steps are:

[0057] The E.coli BL21 original bacteria were inoculated into sterilized LB medium, cultured at 30°C and 170rpm for 24h, and then centrifuged at 10000rpm for 10min to collect the bacteria. Inoculate the E. coli BL21 engineered bacteria prepared in Example 1 into HB-PET self-inducing medium, culture at 30° C. and 170 rpm for 24 hours, and then centrifuge at 10,000 rpm for 10 minutes to collect the bacteria. Among them, the self-inducing medium was purchased from Qingdao Hi-Tech Park Haibo Biotechnology Co., Ltd., and 1 L of the medium contained 27.5 g of culture powder and 5 g of glycerol.

[0058] (2) The preparation of Pt(II) stock solution, concrete steps are:

[0059] 0.864g PtCl 2 Diss...

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Abstract

The invention discloses a surface display metal binding protein gene and application thereof to recycling of platinum and palladium metal. A nucleotide sequence of the surface display metal binding protein gene is shown as SEQ ID NO. 3 and an amino acid sequence of the surface display metal binding protein gene is shown as SEQ ID NO. 6; a recombinant genetically engineered bacterium containing thegene is applied to the recycling of the platinum and palladium metal of industrial wastewater. According to the surface display metal binding protein gene disclosed by the invention, an EC20 gene andan InaKN gene are connected together through an expression vector pET28a(+) by utilizing properties of anchoring protein and are introduced into a microorganism receptor and are subjected to recombinant expression; in a recombinant expression process, EC20 is expressed and fixed on the surface of a microorganism through InaKN, so that the selectivity and the adsorption performance of the microorganism on the platinum and palladium metal are improved; the adsorption capacities of the microorganism on the platinum and palladium metal ions are increased by 1.6 times and 1.3 times respectively, and the recovery rates of the microorganism on the platinum and palladium metal ions in the industrial wastewater reach 100 percent to 97.5 percent respectively.

Description

technical field [0001] The invention belongs to the fields of genetic engineering and precious metal recycling, and in particular relates to a surface-displayed metal-binding protein gene and its application in recycling platinum and palladium metals. Background technique [0002] Due to their special physical and chemical properties, platinum group metals are widely used in various fields, such as industry, agriculture, medicine, catalysis and other fields. With the advancement of science and technology, the use of platinum group metals is becoming more and more extensive, and the demand is also increasing. However, its content in the earth's crust is very low, and the reserves of platinum group metals in my country are less than 5%. Under such a situation, from The recovery of platinum group metals from secondary resources has certain social significance and economic value. At present, the methods for recovering platinum group metals from industrial wastewater are mainly c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/62C07K19/00C12N15/70C02F3/34B01J20/24B01J20/30C02F101/20
CPCB01J20/24B01J2220/4868C02F3/34C02F2101/20C07K14/825C07K2319/035C12N15/70
Inventor 谢建平谭玲刘新星邱冠周
Owner CENT SOUTH UNIV
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