The safety of our project consists of two aspects. One is the laboratory safety; the other is the safety of spraying Pantide into the environment.
Before entering the laboratory, all the members in NCTU_Formosa must receive several training courses and evaluate exams offered by Environmental Protection and Safety Center. The content of the training courses consists of four parts.
- For the basic laboratory safety instruction, we learned the safety level of laboratories and personal protective equipment.
- For the laboratory emergency response, we learned of how to react to several emergency situations and certain procedure dealing with specific chemicals.
- For the toxic chemical operation and management, we learned the content of the material safety data sheet (MSDS).
- For biosafety, we learned biological hazard, including levels of biohazard and instructions of dealing with biological trash.
The four training courses contain an evaluating exam. After taking the courses and passing the exam, we all get certificates for each training.
Safety of Pantide
Processing Procedure in the Lab
In the future, we plan to spray Pantide onto the environment. Therefore, we should guarantee that Pantide is safe to humans and harmless to the environment. As a result, we design a processing standard in the laboratory.
- Free of E. coli
The product we spray into the environment should not contain any genetically-modified organisms. So, after we expressed Pantide with E. coli, the LB broth containing E.coli should be first sonicated by the sonicator to let Pantide be soluble in the LB broth. In this step, most of the living E.coli will be eliminated. In order to guarantee the sonicated LB broth lysate is free of E. coli, we boil the lysate for 1 hour. However, boiling the solution may induce Pantide degradation. We want to know the degradation time of boiling the lysate. Pantide are retained and we run SDS-PAGE to evaluate the amount of Pantide residue. (Figure. 1)
Figure1. SDS-PAGE gel and the concentrations of boiling test to native Hv1a-lectin (17.1 kDa). The marked band shows that there is a significant decrease in Hv1a-lectin after 2 hours.
- Degradation of Pantide
To ensure that Pantide will degrade into amino acids when it is released into the environment, we have done an experiment to prove that Pantide does degrade in the ultraviolet light. (See the experiment) Once we have known that Pantide will degrade in the ultraviolet light, we may conclude that Pantide will degrade in the natural light over time. With the degradation modeling simulation (See the Modelling), we may also apply the degradation rate into the automatic spraying device.
For humans, Pantide will not have any negative effect on human health. When the customer gets the agricultural produce from the market, most of the Pantide is thought to have degraded into amino acids and even there is some residue of Pantide on the surface of the product. Pantide is also harmless to humans. The three toxins we choose have been proved to be ineffective to mammals due to the structural difference between the voltage-gated ion channels between mammals and insects. It has been validated through references doing experiments on mice.
For the environment, Pantide is made of amino acids. So it is harmless for the environment. Instead, it can be as the fertilizer. When it is washed into the soil or in water resources, Pantide is ineffective to the microbiomes and the vertebrate in the ecosystem. Furthermore, Pantide is species-specific, so it will not harm the beneficial pollinator, Bees.