Partnership

LZU-HS-China-A used carrier protein and cell surface display technology to improve the output efficiency during the experiment. Therefore, when looking for partners, we found LZU-HS-China-C and BFSU-ICUnited, which also used carrier protein and cell surface display technology.

Partnership teams

LZU-HS-China-A：LZU-HS-China-A found that a large amount of shrimp and crab shell waste is generated every year, and the team believes that this waste can better play their role. LZU-HS-China-A attempts to convert chitosan into chitosan oligosaccharides by cell surface display.

BFSU-ICUnited：Our team BFSU-ICUnited, is a high-school team united from many regions in China. ICUnited has initiated attempts to tackle one of the most massive environmental issues by offering a novel method for BPA detection which features the use of electrochemical enzyme biosensors.

 LZU-HS-China-C：LZU-HS-China C's project focused on preventing the inherent hazards of antibiotic release due to the improper disposal of polluted wastes in the poultry business, namely the potential of superbugs. They went on to design a whole-cell biocatalyst aimed at degrading antibiotic residuals in animal feces. Their team needed cell surface display technique to improve their otherwise inefficient antibiotic degradation rate.

Cooperative project

based on the selection of anchoring proteins

Since we all use cell surface display technology in our projects, we can help each other with the related problems of this technology. The common problem we faced in the early days was how to find an anchoring protein suitable for team projects. It was a key part of our three-team project, but we didn't know which protein to use, so in our first online meeting, we made a detailed plan for the future progress of our project.

We initially did some research based on the required properties of anchoring proteins Three teams, LZU-HS-China-ALZU-HS-China-C and BFSU-ICUnited, found six articles about INP:
1.Development of Bacterial Cell Surface Display Systems Using Ice Nucleation Protein Anchor Towards Biotechnological Applications
2.Redesigning of Microbial Cell Surface and Its Application to Whole-Cell Biocatalysis and Biosensors
3.Ice-nucleating proteins are activated by low temperatures to control the structure of interfacial water
4.Surface Immobilization of Human Arginase-1 with an Engineered Ice Nucleation Protein Display System in E. coli
5.The structure and mechanism of the TolC outer membrane transport protein
6.Membrane transport protein
Three teams selected two articles each and wrote a report. We made a report at the joint meeting of the three teams and shared such a meeting, which made the three teams understand the anchoring protein more clearly and obtained the following information:
1. The desired properties of anchoring proteins are signal peptide sequences or transport signals that guide fusion proteins through cell membranes.
2. The fusion protein can be directed to a specific site on the cell surface.
3. Stable anchoring elements can be recognized at specific positions on the cell surface so that the fusion protein can be immobilized on the cell surface.
4. Fusion with foreign proteins does not change its stability and it is not easily decomposed by proteases inside and outside cells.

In addition, we also analyzed the anchoring protein. Through analysis, we found that the structure of anchor protein and guest protein easily interacted with each other, interfered with protein folding, and reduced guest protein activity and different transporters affected different physiological functions of receptor bacteria. If inappropriate proteins were used, it might lead to instability of bacterial growth and so on. Eventually, we decided to use ice nucleoprotein as an anchoring protein because it can be expressed stably in a variety of host cells and is not easily decomposed by proteases inside and outside cells. At the same time, INP can be displayed efficiently on the surface of host cells, and its repetitive structure can be cut under certain conditions without affecting the experimental results. This characteristic can ensure that INP does not need other auxiliary proteins when carrying guest proteins with larger molecular weights, which is beneficial to reduce the complexity of experiments.

Publicity

The three teams set up their social media: “IC有nited”, “EcN IL养成记2022篇” and “iGEM虾兵蟹将”. After the first meeting, we decided to let more people know about us by promoting each other's projects. We achieved this goal in two ways. First, we summarized other people's project descriptions to make them simpler and published them in their official accounts. Although we only have a small number of people to pay attention to, the total number of followers of our three teams is still relatively large. On their respective WeChat official account, the three teams introduced their teams and projects and sent them to the heads of each team in the cooperation group, and forwarded the WeChat official account of the other two teams to help each other for publicity.

Education

In preparation for the educational part, the three teams once worked together to contribute to the general public's synthetic biology. We assign work through online meetings. BFSU-ICUnited is responsible for presenting LZU-HS-China-A is responsible for making PPTs and posters, and online lectures LZU-HS-China-C is responsible for preparing related content. This classification maximizes the use of the advantages of the three teams to ensure the efficient implementation of education.

References

Federici, L., Walas, F., & Luisi, B. (2004, July 25). Outer membrane
transport protein - JSTOR. JSTOR. Retrieved October 6, 2022, from https://www.jstor.org/stable/24108864 Han, L., Zhao, Y., Cui, S., & Liang, B. (2017, November 22). Redesigning of microbial cell surface and its application to whole-cell biocatalysis and biosensors - applied biochemistry and biotechnology. SpringerLink. Retrieved October 6, 2022, from https://link.springer.com/article/10.1007/s12010-017-2662-6 Pardee, A. B. (n.d.). Membrane transport proteins - JSTOR. JSTOR. Retrieved October 6, 2022, from https://www.jstor.org/stable/1725415 Roeters, S. J., Golbek, T. W., Bregnhøj, M., Drace, T., Alamdari, S., Roseboom, W., Kramer, G., Šantl-Temkiv, T., Finster, K., Pfaendtner, J., Woutersen, S., Boesen, T., & Weidner, T. (2021, February 19). Ice-nucleating proteins are activated by low temperatures to control the structure of Interfacial Water. Nature News. Retrieved October 6, 2022, from https://www.nature.com/articles/s41467-021-21349-3 Zhang, Z., Tang, R., Bian, L., Mei, M., Li, C., Ma, X., Yi, L., & Ma, L. (2016, August 1). Surface immobilization of human arginase-1 with an engineered ice nucleation protein display system in E. coli. PloS one. Retrieved October 6, 2022, from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4968799/#:~:text
=Ice%20nucleation%20protein%20%28INP%29%20is%20frequently%20used%2
0as,its%20surface%20display%20efficiency%20for%20h
uman%20Arginase1%20%28ARG1%29. 豆丁网. (n.d.). 利用冰晶核蛋白构建细菌细胞表面展示体系及其应用研究. 豆丁网. Retrieved October 6, 2022, from https://www.docin.com/p-701645524.html