Design

Since the current solution to the blockage is not environmentally friendly and economical, we hope to use synthetic biology to solve the three problems of rust deposition, biofilm accumulation and microbial activity by engineering microbe.

However, the three functions of engineered bacteria may bring great metabolic pressure to the engineered bacteria, and the anti-bacterial substances produced by them may adversely affect their growth and reproduction. So we need to design a set of genetic circuit to improve this situation and achieve a sequential expression of these three functions!

Inspired by the way people currently deal with it, people first add rust remover and biofilm remover, which first destroy the living environment of the microorganisms, causing them to fail to receive protection, and then bactericide is added to achieve high bactericidal effect. Therefore, in our gene circuit, it is necessary to perform the function of destroying the microenvironment of the microbial before performing the sterilization function.

To perform the function of destroying the microenvironment, the very first thing is to sense it!

Based on the principles of quorum sensing, we realized the identification of bacterial microenvironment. This enables our engineered E. coli to recognize the quorum sensing signal molecule AHL produced by iron bacteria, activate the expression of biological rust removers and biological biofilm remover. The biological rust remover we chose is the strongest ferric chelating agent-siderophore in nature. As for biofilm remover we chose a glycosidase from the actinomycetemella actinomycete that degrades the biofilm polysaccharide backbone.

With the progress of rust removing, the siderophore-Fe3+ complex concentration will continue to increase, and we hope that after the bacteria sense this signal, the expression of the biocide will be initiated. But in the natural iron sensing system, high concentration of iron inhibit the expression of genes, so we need to modify them. By introducing the foreign gene- lacI, the lacI was inhibited under high-iron conditions, and lacI inhibited the biocide gene, thereby realizing the design of the iron reversal system.

Biocides we tried to use cecropin AD, an antibacterial peptide derived from silkworm, which has a spectral and efficient bactericidal effect. In order to release this peptide to the extracellular, we achieve cell rupture by simultaneously expressing the autolysin gene.

For biosafety reasons, avoid engineering bacteria to escape from the piping system into the natural environment. We think of the dark environment inside the pipeline, and the natural environment is the light environment, so the light-on suicide is adopted, and the suicide gene is expressed when there is light to commit suicide.

For higher biosafety and detectability, we have designed a new hardware-iTube that can be applied to the piping system. ITube's built-in sensors can detect circulating water parameters, use artificial neural networks to conduct corrosion prediction and blockage assessment of cooling water systems, reminding people to add engineering bacteria for plugging and cleaning. At the same time, the built-in light system can also be manually turned on and off, makes the growth of the engineered bacteria in pipe is artificially regulated.

Reference

1. J. B. Neilands (1995). "Siderophores: Structure and Function of Microbial Iron Transport Compounds". J. Biol. Chem. 270 (45): 26723–26726.

2. Nealson, K.; Platt, T.; Hastings, J.W. (1970). "The cellular control of the synthesis and activity of the bacterial luminescent system". Journal of Bacteriology. 104 (1): 313–22.

3. Gilston BA, Wang S, Marcus MD, Canalizo-Hernández MA, Swindell EP, Xue Y, Mondragón A, O'Halloran TV (Nov 2014). "Structural and mechanistic basis of zinc regulation across the E. coli Zur regulon". PLoS Biology. 12 (11): e1001987.

4. Boman HG (1991), "Antibacterial peptides: key components needed in immunity", Cell, 65 (2): 205–207,