Team:UC Davis/Safe
Safe By Design
Cyantific has prioritized food safety in every step of designing and producing our pigment:
- There are two sources of risk we address here: 1) Fundamental health risks (general toxicity, allergenic nature) 2) Dietetic and social/sociological public health risks of a dye like ours.
- As our dye allows us to experiment with the framework of governmental food science policy, it is crucial that we carefully consider the literal toxicity threat inherent in any food dye, as well as the social implications of releasing our protein based dye.
- We are aiming to meet a variety of standards to meet and exceed global health expectations. For instance, in the US we will be trying to meet FDA standards, and in other countries we will be working with their stipulated standards to attend to the nature-identical provisions in the European Union, as well as observing other market’s standards for genetically altered food product in the age of precision agriculture.
- In the future, our ultimate vector will be a Generally Regarded As Safe (GRAS) organism so that regardless of original biological origin of the protein, the entire cell would be safe for consumption, although we have worked out purification methods that yield industry-standard levels.
Cyantific has prioritized food safety in every step of designing and producing our pigment. As our dye allows us to experiment with the framework of governmental food science policy, it is crucial that we carefully consider the literal toxicity threat inherent in any food dye, as well as the social implications of releasing our protein based dye. We are aiming to meet a variety of standards to meet and exceed global health expectations. For instance, in the US we will be trying to meet FDA standards, and in other countries we will be working with their stipulated standards to attend to the nature-identical provisions in the European Union, as well as observing other market’s standards for genetically altered food product in the age of precision agriculture.
We intentionally chose an ultimate vector to be a Generally Regarded As Safe (GRAS) organism so that regardless of original biological origin of the protein, the entire cell would be safe for consumption, although we have purification methods that do not add any foreign DNA bases. Provided that all of our testing shows that it is safe, such purification would not be an absolute necessity. There is no regulatory purity requirement; we have worked so that after we achieve expression of the protein in Bacillus subtilis we can place the entire organism into a food product. There may be some advantages to this approach, as we learned from our interview with Dr. Bruce German, who studies human-bacteria interactions to better understand how our microbiomes contribute to improved health. For more on potential full-bacterium application scenarios, please check the Application Scenarios portion of our Synergy with Synenergene page.
We have implemented safety practices from the ground up. We consulted Dr. Belinda Martineau, a scientist involved with the first mass-marketed genetically engineered food, the FlavrSavr Tomato©, who currently is a member of the UC Davis Institute for Social Science. She detailed the processes she traversed with the trailblazing organism. She urged us to implement testing even beyond what the government regulatory agency requires, as her experiences with agrobacterium yielded results that were unanticipated, specifically additional gene transfer for ampicillin resistance. Because of her experiences at CalGene and with her generation of synthetic biology she has been involved with GMO labeling initiatives with an educated scientific perspective. Since technology has progressed and we are able to use golden-gate transformations and other methods for creating our transgenic plasmids, we do not have to face this specific consequence, but we are circumspect in our processes to head the very real health concerns emerging from new technologies.
There are two sources of risk we address here: 1) Fundamental health risks (general toxicity, allergenic nature) 2) Dietetic and social/sociological public health risks of a dye like ours.
- Chemical Heath Risk
- Organism-Generated Health Risk: This may be mute as we are intent on putting the gene into a GRAS organism and have practiced purification methods to eliminate any contamination concerns, beyond the requirements of applicable regulation. Nevertheless, the protein producing the blue pigment is not currently in the human food system, and that means we must exercise caution. The protein is a cyanobacteriochrome, or CBCR, which is the phytochrome-like complex in cyanobacteria. Although it was not in the scope of our project to determine and isolate the species the protein comes from, the cyanobacteria genera encompasses a range of species with differential effects on human health. It is unclear at this time whether some of the CBCRs we are using have ever been eaten by humans which makes it very challenging to predict exactly what health risks may be present. Multiple sequence alignments suggest that some CBCR’s come from species that have never been eaten by humans, and others come from clades closely related to Spirulina, which is an FDA approved food dye already. Further studies on the origins of the proteins need to be performed in order to determine which species do not pose a human health concern.
- Testing for Human Consumption
- Risk Identification: Our team has identified additional risks to consider and we have studied the specific regulations of the United States as well as a general context. Our protein has likely not been consumed by humans before. This itself instigates substantial testing. Conveniently, proteins are rarely toxic, and a common sign of toxicity is proteins which are stable beyond this threshold. This is why a significant number of experiments to prove safety as a voluntarily selected “supplement” would be in order, but as a coloring agent in human foods, the safety threshold increases. Going forward, our plan is to explore 1) denaturation tests to confirm that the protein is not an allergen in humans, and 2) high volume toxicity studies, though we don’t expect these to indicate any increase in harm given the organic nature of our dye, unlike its’ coal-tar counterpart.
- Denaturation Testing: To ensure this is safe in every possible direction we have to determine that this protein is not an allergen. Therefore, we conduct denaturation testing to determine whether the protein denatures at certain acidity within the human digestive tract. Optimally, the protein maintains its shape through a relatively wide range of alkalinity required by varying production operations involved in making processed foods, but breaks down at pH 1.5 or 2—the acidity of pepsin in the stomach. Even if it breaks down at a higher threshold, so long as the broken down protein is effectively inert and itself is determined to be non-allergenic and safe for waste treatment plants, then this risk is mitigated.
- High Volume Toxicity Studies: Although this would not be for at least a few years from the scope of this project, food colorings are expected to reach a very high threshold for nontoxicity in order to be considered socially worthy to be allowed on the market. Because the marginal utility of having brighter foods as augmented by coal-tar dyes is beneficial primarily to the consumer, even very remote carcinogenic potential makes a dye against the broader public interest.
- Risk Identification: Our team has identified additional risks to consider and we have studied the specific regulations of the United States as well as a general context. Our protein has likely not been consumed by humans before. This itself instigates substantial testing. Conveniently, proteins are rarely toxic, and a common sign of toxicity is proteins which are stable beyond this threshold. This is why a significant number of experiments to prove safety as a voluntarily selected “supplement” would be in order, but as a coloring agent in human foods, the safety threshold increases. Going forward, our plan is to explore 1) denaturation tests to confirm that the protein is not an allergen in humans, and 2) high volume toxicity studies, though we don’t expect these to indicate any increase in harm given the organic nature of our dye, unlike its’ coal-tar counterpart.
- Safety on the Market—A Closer Look at FDA Policy: The international regulatory landscape around food additives is complex. For instance, the US has The logic underlying this policy is that, since coloration of food (historically making food producers liable for fraud and a major source of consumer poisoning) is for strictly vanity reasons and visual appeal, there is no excuse for a food colorant to be even slightly carcinogenic or otherwise harmful. The benefit cannot exceed the risk unless the risk of the dye’s use is exceedingly minimal. This involves the aforementioned test, but also includes a long review process with rat studies, among others. The required threshold is one in one million deaths. After this work, the dye would have to fit into a category (natural, like Carmine from beetles; SynBio’s beta-carotene; or artificial, like coal-tar dyes). Among these categories are separate lists under the Food, Drugs, and Cosmetic acts detailing exactly which uses are appropriate. For instance, Spirulina, a relative of the bacteria where our CBCR’s come from, has very specific approved uses in certain candy products, for instance, but not as a general dye—coating insufficiently ripe blueberries, for instance, would be a violation. These lists are further sorted into provisional and permanent—although the provisional list is very stable and colorings are somewhat rarely dropped. <br/> Our dye is best imagined alongside beta-carotene, as this is a nature-identical dye produced originally in foods such as carrots. The main distinction that would contest this placement in the natural category is that the original producer of beta-carotene has been used for its color before—carrot juice has long been among the Parthenon of food colorants; our cyanobacteria source has likely not. Other than this contestation, there are no naturally occurring substances (such as a protein like ours) on the artificial lists, and there is no reason other than the interference of this iGEM team in the bringing forth of this bright protein to suggest such a label. In the binary of the food additive complex, it is highly likely that the FDA would determine our extract to have a “natural coloring” label. Whether this is the most ethical outcome is explored elsewhere on the wiki.
- Dietetic Concerns
Now, in regard to the social welfare of the dye, the principle concern would be that if our dye product is used considered a natural dye, it will enable processed foods to be marketed as “more natural” when in fact they may still contain low nutritional content. This labeling issue is addressed through our human practice research and we also contend that, as our dye is a protein and not a coal-tar color (which are small molecules that cannot be digested), there is some plausible health benefit to replacing the artificial coloring with our biological one. As proteins break down into amino acids, they do carry some nutritional content. And so long as they merely replace the less healthful dyes and do not produce new markets for even more nutritionally weak foods, then we consider our dye to have some marginal benefit for the consumer—even if they are unconscious of the effects. For further analysis of possible sociological affects, positive and negative, please check out the Seeing Blue: Scenarios for Synenerge page.
