Engineering strain for controlling adsorption and escape of heavy metals as well as construction method and application of engineering strain
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A technology of heavy metal ions and bacteria, applied in the field of genetic engineering, can solve the problems of staying in the theory of academic literature, low adsorption efficiency, secondary pollution, etc.
Pending Publication Date: 2022-02-01
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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However, the adsorption efficiency of heavy metals based on microorganisms is not high, and further modification is needed to improve the activity; on the other hand, all the heavy metals that are adsorbed still exist in the environment, and there is a risk of secondary pollution. It is urgent to develop a fast , low-cost recycling means; finally, biosafety
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Embodiment 1
[0075] Example 1, the design of water heavy metal adsorption and escape control microorganisms and the optimization of each functional element
[0076] (1) Design of water heavy metal adsorption and escape control microorganisms
[0077] 1. Heavy metal responsive promoter and relevant information
[0078] Through literature research, heavy metal-responsive promoters and relevant information are shown in Table 1.
[0079] Table 1. Heavy metal responsive promoters and related information
[0080]
[0081] 2. Heavy metal adsorption, bacteria enrichment and escape control microbial gene circuit design
[0082] The heavy metal ion adsorption, cell enrichment and escape control microorganism designed in the present invention comprises a heavy metal ion response element, a heavy metal ion adsorption element, a cell enrichment element and an escape control element. Taking mercury ion as an example, the composition of each component is as follows:
[0083] The mercury ion respon...
Embodiment 2
[0196] Example 2. Construction and functional verification of mercury ion adsorption, bacteria enrichment and escape control engineering strains
[0197] 1. Construction of mercury ion adsorption and escape control engineering strains
[0198] Utilize the optimized functional elements in Example 1 to construct engineering strains, the specific steps are as follows:
[0199] 1. Construction of recombinant vector
[0200] 1) Based on the medium copy plasmids pRG (p15A replicon) and pUCP20 (ColE1 replicon (E.coli), pRO1600 replicon ( P.putida)), construct two shuttle vectors p2015E and p2015P (vector map such as Figure 6 shown), containing the optimized mercury ion-induced promoters in Escherichia coli and Pseudomonas putida, respectively, and the enzyme cutting sites Spe I and Pst reserved for connecting the protein gene sequences of the adsorption, recovery and escape control elements I.
[0201] The nucleotide sequence of the p2015E vector is shown in sequence 15, wherein...
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Abstract
The invention discloses an engineering strain for controlling adsorption and escape of heavy metals as well as a construction method and an application of the engineering strain. The engineering strain is a bacterium obtained by introducing a complete system into a host bacterium; the complete system comprises a heavy metal ion regulation protein, a heavy metal ion adsorption protein displayed on the surface of an ice nucleation protein, a streptavidin binding protein displayed on the surface of the ice nucleation protein and an escape control element, and the escape control element comprises repressor proteins PhlF, TetR, LacI and a lethal protein. Based on a synthetic biological technology, the invention provides a water body heavy metal adsorption and escape control microorganism which integrates the functions of heavy metal adsorption, microorganism enrichment after adsorption, escape control and the like in escherichia coli or pseudomonas putida, and the construction of a complex, fine and predictable genetic loop is realized by utilizing a standardized biological element; after optimization design, gene lines of adsorption, enrichment and death behaviors can be completed under the condition of mercury ions with different concentrations.
Description
technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a heavy metal adsorption and escape control engineering strain and its construction method and application. Background technique [0002] Hg, Cd, Pb, etc. in the natural environment will have adverse effects on the growth of organisms in nature, and will eventually threaten human health under the enrichment of the food chain. The direct discharge of industrial pollutants without strict treatment of heavy metal components has caused very serious pollution to some water bodies. Therefore, the cleanup of heavy metals in water bodies has become a major environmental problem that needs to be solved urgently. [0003] Some microorganisms in the natural environment participate in the biochemical cycle of heavy metals and become an important part of it. These natural microorganisms are an important link in maintaining the balance of mercury in the environment. U...
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