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Project Results

Figure 1. Fluorescence per cell for different glucose levels for the four promoters (Edd, Kgu, Gad, Zwf)

Figure 2. Quantitative image processing of a confocal microscopy image (Alginate encapsulated P. Putida (Gad) at 10% glucose). From left to right: brightfield image, fluorescence image, and analyzed image.

Figure 3. 3D view of our glucose-sensing Pseudomonas Putida in an alginate-based capsule (300 µm × 300 µm) obtained by confocal microscopy and processed in ImageJ.

Improvement over the state of the art

Our glucose sensor is the only, and first ever, sensor systems that directly detects and responds to glucose levels based on synthetic biology methodology. Currently, nurses and medical doctors in hospitals use optical-based sensors for measuring glucose levels in diabetics. Previously, the 2011 iGEM team of Missouri

made an indirect glucose detection system, which is based on detecting changes in osmolarity caused by changes in glucose levels. Since our proposed glucose sensor is based on direct sensing of different glucose concentrations, it has the potential for is a more precise measurement technique than both the optical sensor and the system of the Missouri iGEM team.

The optical sensing system that is used in hospitals is typically applied twice a day on a patient. This is not frequently enough to get a good handle on glucose-level changes in patients. Our system is designed to continuously measure and report the glucose levels. Moreover, upon detection of a critical concentration of glucose, our proposed system will begin to secrete insulin at a dose commensurate with the length of time the supercritical glucose levels are present. Currently at hospitals after detection of glucose with the optical system, the same dosage of insulin will be injected to different diabetic persons with different levels of glucose in their blood.

Consequently, the use of our engineered P. putida strain as a glucose sensor should be considered as a more sensitive method than the existing optical sensor. Additionally, it is not a time consuming procedure, since after its implantation a diabetic person does not need to get a daily care by doctor or nurse. Therefore, by decreasing the workload of medical personnel and physical equipment, the cost of glucose sensing is expected to drop. It is conceptually possible to implant the bacteria since they are encapsulated in Alginate, which renders the product untriggered by the immune system of the patient. After implantation, our encapsulated P.putida will last for a long period, so that is clearly a positive point and improvement compared to the optical system which requires frequent access and controls. Finally, a functioning implementation of our proposed system has the potential of significantly suppressing the symptoms of diabetes, thus making it possible to live as a non-afflicted person.

In addition, this make it as a more convenient method for diabetic people because they can most likely live as a normal person without diabetes.

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