Team:Fudan/Poster

Poster: 2020 Fudan

Make calcium supplement sustainable

Bone appetite
  • Student Leader: Shizheng Tian
  • Student Members: Shuojin Liang, Mingwei Li, Ziyuan Wei, Jingqi Zhang, Tianhui Xu, Zhenru Zhang, Yiyang Yao, Yiyang Lu, Yuanyuan Liu, Emily Mo, Xiaohui Jiang, Xinran Li, Gaochen Zhang
  • Advisors: Liang Cai, Yijie Pan, Yuchang Lee, Jingyi Hu
  • Team PI: Daru Lu
  • Affiliation: School of Life Sciences, Fudan University; with funding from YF Capital

In Asia, many old people suffer from insufficient calcium intake and related diseases such as osteoporosis. Considering that traditional calcium supplementation is not efficient and user-friendly enough, we intended to develop a kind of engineered bacteria based on E. coli that can colonize in the human intestine and secrete peptides to promote calcium absorption.

We introduced the quorum sensing system to colonize bacteria in the intestine. Once the bacteria have successfully colonized the intestinal tract, the luxPR promoter will be activated and a series of short peptides that can bind to calcium and promote calcium absorption of intestinal epithelial cells will be expressed. A kill switch is added accordingly for safety.

Our project provides a novel delivery method to provide calcium supplements sustainably. And we are actively seeking commercialization possibilities to expand our project impact, to reach more population, for the elderly's good health and well-being.

So far, what we have achieved

项目缘起，需要HP那四张关于骨质疏松的问卷图，数据不变 重画

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铜奖满足，但不是罗列因为评委有一个列表给奖牌打分

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银奖满足，但不是罗列因为评委有一个列表给奖牌打分

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金奖满足，但不是罗列因为评委有一个列表给奖牌打分

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  • We have zzzz bala bala bala ba la ba la
  • We have wxcv bala bala bala ba la ba la

总结的话

SDG 3: GOOD HEALTH AND WELL-BEING

描述我们项目如何贡献SDG3；为什么关怀老人，我们完成了什么，后来人继续做什么 bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la

Our project aims to provide a holistic solution to the problem of osteoporosis among seniors. Our online general biology course Shengyin and senior-friendly textbooks, are also designed to educate the elderly on the importance of bone health.

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SDG 4: QUALITY EDUCATION

描述我们项目如何贡献SDG4；我们为什么要教育，完成了什么，后来人继续做什么 bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la

Our free online audio course The Voice Of Biology runs on multiple popular platforms with easy access to all age groups. We incorporated graphics to attract younger audiences and provided large font textbooks to accommodate seniors and those with hearing impairment. 上传两个ogg格式的文档

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bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la What is the problem your team is working to solve? How does it affect the world?

SDG 9: INDUSTRY, INNOVATION AND INFRASTRUCTURE

描述我们项目如何贡献SDG9；我们完成了什么，下一步将要做什么 bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la

We created a thorough business plan for our project and further explored possible innovation in the industry by participating in youth innovation competition, pitching to investors and much more.

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bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la bala bala bala ba la ba la How are you going to solve the problem? Where did the idea come from?

We invented CaAP

CaAP stands for Calcium Absorption Peptide.

According to literature, we selected peptides @@@@GPAGPHGPVG, FDHIVY, YQEPVIAPKL, NDEELNK, and DHTKE. All of the first four peptides can form calcium-peptide complex through their Asp and Glu residues, and facilitate cell uptake. DHTKE uses its Asp, Glu and His residues to do the same. Basic part @@@@

We add a signal peptide to the N segment of CaAP to guide its secretion into the periplasmic space of E. coli. Because it is so small, it will then leak out of the cell wall into the outside environment. @@@@ 2 existing parts, 3 new; composite parts

We linked these peptides through Phe-Arg (FR) junctions. When CaAP is expressed and secreted by the engineered E. coli in the intestinal environment, CaAP could be further cleaved into functional oligopeptides in the presence of digestive enzymes (trypsin and chymotrypsin) in the intestinal lumen.

We improved mcbABCD
Based on previous work, We decided to use luxI/luxR and TetR to construct a quorum sensor system with the antimicrobial peptide microcin B17 (MccB17) expression system to achieve the "on" and "off" effects of controlling the growth of engineered bacteria. McbABCDEFG jointly completes the expression, processing, and secretion of MccB17. Among them, mcbA expresses the main structure of the antimicrobial peptide, and mcbBCD is responsible for the post-translational modification of the antimicrobial peptide. McbEF expresses the channel protein to secrete the peptide antibiotic. McbG is accountable for the immunity against MccB17. McbEFG is all related to E. coli 's immunity while self-producing microcin. TldD/tldE is a metalloprotease that is ubiquitous in bacteria. When the surrounding bacteria absorb MccB17 produced by Escherichia coli , TldD/tldE will activate it and kill the bacteria. In our antimicrobial peptide expressing system, McbABCD is under the control of a constitutive promoter, but PtetR regulates the expression of McbEFG. When the number of engineered bacteria in the intestine is small, few dimers formed cannot activate the luxpR promoter. Thus, TetR and CaAP expression systems are both silent. However, McbEFG is expressed and transports MccB17, which can inhibit the growth of surrounding bacteria. When the number of bacteria increases to a certain extent, the dimers formed by the products of luxI and luxR will accumulate and activate luxpR. At this time, tetR is turned on, and PtetR is off. As a result, McbEFG stops expressing, and the CaAP system begins to work. Since the population number has reached a certain level, the engineered bacteria can express CaAP more efficiently. Meanwhile, MccB17 is still continuously expressed but cannot be effectively effluxed, which will cause the toxins to gradually accumulate in the engineered bacteria and prevent them from continuously unrestricted proliferation. To achieve better expression efficiency, we have made an improvement in this system by creating a new promoter fusion. As ptetR has been studied thoroughly and is well-characterized, we created promoter fusion of pl and tetO. Not only is it regulated by tetR and anhydrotetracycline, but it also inherits the expression level of pl. This also allows us to create the desired promoter simpler. @@@@ To solve the problem, we have proposed some improvement plans. By researching the literature and talking with Professor Lin from the Fudan University Academy of Sciences, we decided to use low-copy plasmids to reduce the expression burden of engineered bacteria. We tried to keep the main part of MccB17 (McbABCD) a relatively low expression and high express the immune part (McbEFG) to achieve high secretion of MccB17. The highly expressed channel proteins and immune proteins will export more MccB17 and reduce the accumulation of antimicrobial peptides in the engineered bacteria cells, thereby inducing them to produce more antimicrobial peptides.
CaAsst for measuring trace calcium ion
Traditional blood calcium testing requires at least 5ml blood. We developed a new measurement technique called CaAsst. We use methylthymol blue to convert the measurement of calcium concentration into the measurement of absorbance. Experiment results show a linear relationship between calcium concentration and absorbance within a specific range. We also introduced some interference substances to showcase the stability of our measurement project. CaAsst has more room for development and may be widely adapted for other tests like the blood sugar test.
How to use our parts
What did your team achieve? What do you plan to work on moving forward?
Acknowledgements
If not already cited in other sections of your poster, what literature sources did you reference on this poster? Who helped or advised you?