Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Recombinant bacillus subtilis capable of efficiently adsorbing multiple heavy metals as well as preparation method and application of recombinant bacillus subtilis

A Bacillus subtilis and heavy metal technology, applied in the direction of biochemical equipment and methods, applications, chemical instruments and methods, etc., can solve problems such as limited effects, achieve strong operability, reduce harm to humans, and be simple and quick to prepare Effect

Pending Publication Date: 2022-05-06
两山生态科技(山东)有限公司
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the field of heavy metal adsorption and passivation, although microorganisms play a certain role, the effect is limited, so it is urgent to develop new microbial adsorption heavy metal technology

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Recombinant bacillus subtilis capable of efficiently adsorbing multiple heavy metals as well as preparation method and application of recombinant bacillus subtilis
  • Recombinant bacillus subtilis capable of efficiently adsorbing multiple heavy metals as well as preparation method and application of recombinant bacillus subtilis
  • Recombinant bacillus subtilis capable of efficiently adsorbing multiple heavy metals as well as preparation method and application of recombinant bacillus subtilis

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0034] The present invention also provides a method for preparing the above-mentioned recombinant Bacillus subtilis that efficiently adsorbs various heavy metals, comprising the following steps:

[0035] (1) Cloning the nucleotide sequence encoding the heavy metal adsorption fusion protein into a plasmid vector, and constructing an integrative plasmid that expresses the heavy metal adsorption fusion protein;

[0036] (2) Linearize the integrated plasmid expressing the heavy metal adsorption fusion protein with linearization enzyme and remove the 5' cohesive end with phosphatase, then transfer it into Bacillus subtilis competent cells for homology Recombination, the positive clone obtained by screening is the recombinant Bacillus subtilis that efficiently adsorbs multiple heavy metals.

[0037] In the step (1), the plasmid vector can be any suitable plasmid vector, preferably, in the step (1), the plasmid vector is a PDL plasmid.

[0038] In the step (2), the linearizing enzym...

Embodiment 1

[0041] Example 1 Recombinant engineering bacteria Bacillus Subtilis WB600 / CotX-P HM and Bacillus Subtilis WB800 / CotX-P HM preparation

[0042] 1. Gene and Primer Preparation

[0043] According to Bacillus subtilis CotX protein (Genbank No.: NP_389058.1), the heavy metal lead highly efficient and specific binding polypeptide P pb (Rui Nian, Duck Sang Kim, et al.2010. Chromatographic biopanning for the selection of peptides with high specificity to Pb 2+ from phage displayedpeptide library[J],Journal of Chromatography A,1217(2010)5940-5949), heavy metal cadmium efficient and specific binding peptide P cd (Mejare M, Ljung S, Bulow L, et al. 1998. Selection of cadmium specific hexapeptides and their expression as OmpA fusion proteins in Escherichia coli [J]. Protein Engineering, 11 (6): 489-494), heavy metal nickel efficient specificity binding peptide P Ni (Li H, Dong W, Liu Y, et al. 2019. Enhanced biosorption of nickel ions on immobilized surface-engineered yeast using nic...

Embodiment 2

[0054] Example 2 Recombinant engineering bacteria adsorption heavy metal cadmium test

[0055] Adopt the Bacillus subtilis culture method of embodiment 1, to cell Bacillus Subtilis WB600 / CotX-P HM 、Bacillus Subtilis WB800 / CotX-P HM , Bacillus Subtilis WB600 and Bacillus Subtilis WB800 were cultured respectively, then the bacterial liquid was centrifuged, the supernatant was removed, and then the precipitate was rinsed with sterile water three times to obtain 0.3 g of each bacterial cell. Take 12 1000mL shake flasks, which contain 200mL of Cd with a concentration of 100mg / L 2+ Solution (pH 6.0), 3 replicates per cell, 0.1 g of bacteria in each shaker flask. The shake flask was adsorbed at 30°C and 200r / min for 120min. After the end, the Erlenmeyer flask solution was centrifuged at 8000r for 10min, and the supernatant Cd 2+ The concentration was measured using a flame atomic absorption spectrometer. Solution Cd before adsorption 2+ The concentration is C 0 , solution Cd a...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides recombinant bacillus subtilis capable of efficiently adsorbing various heavy metals, which is integrated with a nucleotide sequence for coding a heavy metal adsorption fusion protein, and the heavy metal adsorption fusion protein is expressed through the nucleotide sequence. The heavy metal adsorption fusion protein is a fusion protein of a bacillus subtilis CotX protein, a heavy metal lead efficient specific binding polypeptide Ppb, a heavy metal cadmium efficient specific binding polypeptide Pcd, a heavy metal nickel efficient specific binding polypeptide PNi and a histidine cluster polypeptide PHis capable of efficiently adsorbing various heavy metals, and the amino acid sequences are respectively shown as SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8 and SEQ ID NO: 10. Related preparation methods and uses are also provided. The recombinant bacillus subtilis capable of efficiently adsorbing various heavy metals can efficiently adsorb various heavy metals, so that the recombinant bacillus subtilis can be applied to the field of pollution treatment of various heavy metals, the harm of the heavy metals to human beings can be remarkably reduced, and objective economic benefits and social benefits are generated.

Description

technical field [0001] The invention relates to the technical field of heavy metal adsorption, more specifically, to the technical field of heavy metal adsorption bacteria, in particular to a recombinant bacillus subtilis that efficiently adsorbs various heavy metals and its preparation method and application. Background technique [0002] The overall situation of heavy metal pollution in farmland soil in China is not optimistic. The situation of heavy metals in farmland soil exceeding the standard is worrying, and food safety production is threatened. The "National Soil Pollution Survey Bulletin" released in 2014 shows that the rate of soil points exceeding the standard in cultivated land in my country is 19.4%, and the main pollutants are cadmium (Cd), arsenic (As), nickel (Ni) and lead (Pb). The Ministry of Agriculture has 1.4 million hm in my country 2 According to the survey of sewage irrigation areas, 64.8% of the areas with heavy metals in the soil exceeded the standa...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N1/21C12N15/62C12N15/75B09C1/10C09K17/14C09K101/00C12R1/125
CPCC07K14/32C07K7/06C07K7/08C12N15/75B09C1/10C09K17/14C07K2319/00C09K2101/00
Inventor 孙爱友孙冰孙菁泽
Owner 两山生态科技(山东)有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com