Team:SVCE CHENNAI/Scholars
Scholars
To have a deeper insight of our project and the possible social and ethical issues associated with it, the members of our human practices team decided to have discussions with various scientists and scholars in the field of food bio-technology.We also believed that their suggestions will help us in the enhancement of our project,and will also help us to know about the tests and procedures we need to undergo in order to release our final product.We had interviews both through skype, and also in-person.
Out of several scientists and scholars we have talked with,we have listed out :
NAME OF THE SCIENTIST : Dr. Anu Appiah
PLACE : CFTRI(Central Food Technology Research Institute), Mysuru.
WORK STATUS : Principal scientist
SUGGESTIONS AND OPINIONS :
- He wanted us to investigate how proteolytic enzymes in milk will not break our antimicrobial peptide.(suggests to add unusual amino acid so that they don’t degrade the milk)
- Told us how the regulatory system for GMOs in India is very strict and complicated which in turn encouraged us to produce our regulatory report with regard to GMOs.
- He told us how Bacillus subtilis by itself produces protease enzymes, and hence the anti microbial peptide will be broken down within the system.
- Encouraged us to find the dosage required to kill the contaminating organisms., and wanted us to determine the minimum inhibitory concentration.
- Currently for mass transport of milk across states(70000 litres everyday from mysuru to Kerala) , Formalin(very toxic) is used. Encouraged us to device our system such that it will be scalable to an industrial level for the transport of milk.
- Asked us to figure out to make the antimicrobial peptide large enough, that its stable for months together and produce it using fermentation process and isolate it using downstream processing. When we argued plamid expression is less , he told us that we can transfer our plasmid to high expression system such as pichia pastoris.
NAME OF THE SCIENTIST : Dr.Dhyaneshwar B Chawan
PLACE : Liverpool,NY.
WORK STATUS : Owns a food-quality control based company named the SRIM ENTERPRISES.
SUGGESTIONS AND OPINIONS :
- Told us about the precipitant test which is done to screen for the presence of adulterants in milk.
- He suggested us to opt for electron microscopic studies to view our peptide-microbe interaction rather than going for zone of inhibition and spectroscopic studies.
- We were also suggested to monitor the molarity and osmotic property of our medium and were also told to use the control gradient based dialysis tubing as the outer membrane.
- Various propositions were also given by him for the enhancement of our peptides such as Raman spectroscopy to study the structure of peptides.He also insisted us to work on the stability of our peptides at higher temperatures.
- One of the confirmatory tests which is done to identify the presence of GMOs in the sample namely the Q-PCR and Multiplex-PCR (DNA based test) was also suggested by him.
- The pasteurization test,densitometric analysis and the common bio-chemical tests to identify the presence of dipicolinic acid were some of the methods proposed by him for the confirmation of B.subtilis in the medium.
- He told us to perform the heat-shock technique to convert the spores into vegetative form.
NAME OF THE SCIENTIST : Dr.N.Ramaswamy
PLACE : Karur.
WORK STATUS : Managing director and chief doctor at Deepa Kannan Hospital
SUGGESTIONS AND OPINIONS :
- He insisted that the AMPs should not disturb the structural properties of casein and whey (which are the major proteins in milk) as it may have potential health hazards.
- According to him, the term GMO is a “Sharp Double Edged Knife”, and by this,he wanted us to design our product in the most safest way possible.
- He was also concerned about the possibility that our AMPs may trigger cancer cells and the genes responsible for defective birth since he is influenced by the circumstances of genetic disorders caused by GM foods.
- He also suggested us to make sure that our AMPs will not invoke any catalytic activity in expressing the symptoms of several diseases.
NAME OF THE SCIENTIST : Ms.Jaibiba
PLACE : Tiruvallur.
WORK STATUS : Professor of Food-biotechnology.
SUGGESTIONS AND OPINIONS :
- She was concerned about the behavior of our organism at higher temperatures and also about the milk quality on application of AMPs and as well as on the half-life of our peptides.
- She mentioned about the FSSAI and HACCP which are the two main regulatory bodies in India responsible for the approval of our final product.
- The MF membranes made form alpha alumina and the SF membrane made from PVDF were proposed as possible alternatives for the release of 1KDa peptides.
- She expressed her concern about the potential release of metabolites from the GMO into the milk.
- She insisted us to work on the characterization of subtilis strain to make it less pathogenic and also to work on the membranes used in our sachet.
NAME OF THE SCIENTIST : Mr. Abhijeet Gatade(1), Miss.Sini S. Nair(2)
PLACE : Shivaji University, Kohlapur.
WORK STATUS : Assistant Professor(1) ,M.Sc. II Year Student(2)
SUGGESTIONS AND OPINIONS :
- First of all, they applauded us for our efforts and told us that our project is quite interesting and innovative. And they added that it appeared to be novel as it is trying to put forth a solution to the issue of milk spoilage which is of great concern especially in rural areas.
- They felt that the thermo stability of the membrane may become a concern in the construction of our final delivery design (sachet).Thus they suggested the use of high temperature- tolerant membranes in order to resist the spillage of bacteria into the milk as the milk would be heated at a considerably high temperature .
- iii) Suggestions and ideas about some common tests being carried out before releasing a food product was also given by them, which included the microbiological tests, the allergy tests, and the toxicity tests. They insisted the importance of these tests as our project includes many components such as the bacteria, the AMPs and also the membranes. The action of our AMPs against certain harmful organisms causing the spoilage was told to be studied. And they also insisted that attention must be paid on the effect of the AMPs on the useful microbes in milk in order to not destroy them. They also told us to study the effects of our AMPs on the human cells.
- Some common steps to be followed while going for a patent was also given by them, which included the following:
- We are supposed to check for the patentability of our product, and whether it could fulfill the criteria for patenting such as the novelty and industrial applications. After doing all the mentioned steps we would be allowed to prepare our patent draft.
- If we are at the early stage of invention, we can go for provisional specification for about 12 months to file completion and after completing, we can directly go for complete specification. Our patent application will be published after 18 months which is then examined only after receiving request for examination. After the examination we are supposed to give the response to the objections, then clear all the suspicions after which the patent will be granted. It was told that the granting of patent may take upto three years.
- The common concerns during the release of our GMOs may be– susceptibility of non-target organisms, the intervention of peptides and the stability of the gene introduced, and the effects imposed by the AMPs on the human body and as well as the surrounding environment.
- We were suggested to design our peptides in such a way that it could also be utilized to prolong the shelf life of milk during transportation to the industries and other concerned places. It was also advised that care must be taken to ensure that our peptides will not affect the integrity of the milk..
- The most common mode of action of the AMPs which includes the creation of trans-membrane pores followed by cell permeabilization was explained in detail by them, which included the following mechanisms:
- Electrostatic attraction between the cationic peptides and the anionic bacterial cell wall through which the AMPs bind to the LPS (lipo-polysaccharide layer) thereby displacing the divalent poly-anionic cations that eventually lead to the disruption of cell envelope. This mechanism is known as the self- promoted uptake.
- The various models describing about this mechanism includes the Barrel Stave model, Carpet toroidal or aggregate channel model.
- After disrupting the cell wall, the AMPs arrive at the cytoplasmic region where they pass through the interfacial region of target cell to inhibit essential cellular processes.
INFERENCE
From the above mentioned suggestions we can infer that:
- All of them gave us information about the tests and procedures that will be done during the approval of our final product by the respective regulatory bodies.
- Most of them are concerned about the potential hazards that may be posed by our product in the environment.
- They want us to work on the safety of our GMO and the ethical and social issues associated with it.
- Loss of the integrity of milk by our peptides is yet another distress.
- They also fear that our AMPs may have damaging effects On the human system.
- Suggestions were also given to work on the half-life of our peptide and the membranes in our delivery system.
Sponsors
