Team:Thessaly/Proof Of Concept
Overview
Our proof of concept for our project, AMALTHEA, was to combine our successful demonstrations
and Lab results in order to showcase if our project as a whole, will be applicable in the
real world.
2. Capsule administration
We designed a bioelectronic capsule for the detection of gut dysbiosis, through signal transduction. Our non-invasive encapsulated detection device consists of two modules, a genetically engineered bacteria-based module, and an electronic module. The engineered bacteria-based module consists of a bacterial system able to convert the biological signal into an electrochemical one, while the electronic module consists of 3 batteries, a System on a Chip(S.o.C),and a 3-electrode sensor attached in the bacterial tank.
We designed the prototype model of the capsule with the proper dimensions for an ingestible capsule and printed it by a 3D printer into a prototype model with original dimensions for real life implementation.
Figure 1: Front view of the capsule, highlighting the L-arabinose reservoir or “kill switch”.
Figure 2: Pictured here is the final outer cell and components of our capsule.
Figure 3: The 3D printed prototype in actual size (in cm)
Figure 1: Front view of the capsule, highlighting the L-arabinose reservoir or “kill switch”.
Figure 2: Pictured here is the final outer cell and components of our capsule.
Figure 3: The 3D printed prototype in actual size (in cm)
3. Detection of SCFAs
Our engineered bacteria inside the capsule detect the absence of SCFAs and have a reporter module producing a detectable electrical current.
We developed a functional model of the GPCR detection module and predicted the system’s response to various concentrations of SCFAs. We also modeled the Reporter Module’s
function by simulating it with various methods to visualize the final signal it produces.
After the simulation, we expressed Tyrosinase in the lab, responsible for the reporter module. We also showed functionality but these results need repetition.
Figure 1: The GPCR’s output signal (LacI) as the concentration of SCFAs increases.
Figure 2: The results of the deterministic simulation using the ode15s solver, and the according legend showing the color coding.
Figure 3: This SDS page shows the tyrosinase production after induction with 1% L-arabinose.
Figure 4: The ratio 670nm/600nm melanin production/cell growth of bacteria with Tyrosinase assay Buffer
Figure 1: The GPCR’s output signal (LacI) as the concentration of SCFAs increases.
Figure 3: This SDS page shows the tyrosinase production after induction with 1% L-arabinose.
Figure 4: The ratio 670nm/600nm melanin production/cell growth of bacteria with Tyrosinase assay Buffer
4. Signal to smartphone
Our bioelectronic capsule transmits the data collected from the gut to a smartphone to a smartphone.
We assembled a prototype with the use of an Arduino Uno and a bluetooth antenna(HC-05).
The goal of the prototype is to simulate the data transfer via Bluetooth from the
capsule(Arduino) to our smartphone application (e-AMALTHEA). We managed to link and
transfer the data from a datasheet to a smartphone.
Figure 1:The assembled Arduino uno that we used to transfer the data.
Figure 2:The transferred data.
Figure 1:The assembled Arduino uno that we used to transfer the data.
Figure 2:The transferred data.
5. Application
The installation of our application in the user’s smartphone is necessary for the receiving and
visualization of the data, as well as connection with experts. So we created an application which has a Graphical User
Interface and can display the received data.
Figure 1:A) Login at our application ‘’e-AMALTHEA’’. B) How the diagram of SCFAs’ concentration (translated in μV), versus time will look like to a user with a healthy hut. C) How the diagram of SCFAs will look like to a user with gut dysbiosis.
7. Probiotic administration and protection
Along with the personalized nutrition the user should take our probiotic to replenish the
SCFAs missing inside the gut. We had to be sure that our engineered probiotic will reach
and thrive in its target, the large intestine. So we microencapsulated bacteria with
alginate and tested their quality in simulated gastric juice.
Figure 1:Plate on the left: uncrushed alginate beads after treatment. Plate on the right: crushed beads and colonies from the released bacteria.
Figure 1:Plate on the left: uncrushed alginate beads after treatment. Plate on the right: crushed beads and colonies from the released bacteria.
8. Enriched Probiotic
SCFAs production is the main goal of our probiotic in order to contribute to the
amelioration of the problem. So we managed to everexpress to enzymes required for the
production of Acetate, one of the three SCFAs we focus on.
Figure 1:SDS page (L=Lysate S=Souble I=Insoluble)(0%, 0,1%, 1% L-arabinose inducer) Expected bands:Pta 77,2 kDa and AckA 43,7 kDa
9. Probiotic viability and sustainability
After our probiotic is released into the large intestine, we need to be sure that our
probiotic will outcompete the non friendly environment having an advantage compared to the
existing gut flora. So we implemented a module for cellobiose utilisation in order our
probiotic to have an extra carbon source.
Figure 1: Growth (OD600) of E. coli MC1061 cells transformed with N20-BglX (N20), native-peptide BglX (SP) or an empty vector (EMPTY), in a minimal M9 medium with Cellobiose as a carbon source ( CB ).
Figure 2: Growth (OD600) of E. coli MC1061 cells transformed with N20-BglX (N20), native-peptide BglX (SP) or an empty vector (EMPTY), in a minimal M9 medium with Glucose and Cellobiose as a carbon source ( 2 ).