Team:Moscow RF/HP/Silver

Human Practices

The world around us is constantly evolving. Modern science has provided us with perfect tools to adjust the evolution to meet our needs and it is up to us to interfere with the changes or not, to be a player or just a viewer. Synthetic biology and gene engineering has become an essential part of our life even if not accepted by some of us. Primum non nocere. Avoid doing harm. This is the primary and fundamental principle of medicine that can be extended to any field where gene engineering methods are used. Do as much good as you can. This is another principle that could become a part of synthetic biology philosophy. And this is the principle we kept in mind when discussing the future of our project.

The very choice of our project was quite a challenge. In the very beginning we had three project ideas, two of which were about medicines and technological problems of drug design and development. But our final decision was agriculture, and there are two major factors that have determined this choice.

The first factor is quite a global one. It is generally known that the problem of global overpopulation is getting more and more critical, and one of severe consequences of such overpopulation is food shortage and starvation: according to the UNO (Financial Times, Mar. 22, 2017), about 20 million people in Africa have found themselves to be on the verge of starvation in 2017. And there is no doubt that one of potential ways to solve the problem is developing bioengineering technologies making it possible to produce more, faster and less costly.

The second factor is rather a national one. Russia has always been a major producer and consumer of agricultural products, including milk and meat. In spite of the fact that such products account for a significant part of country’s GDP, a huge amount of them is still being imported. However, taking into account the economic restrictions imposed on Russia since 2014 and the general policy of import substitution adopted here in this context, it is obvious that we have all the chances to succeed in providing our population with totally domestic food only in case we stake on advanced manufacturing technologies, including bioengineering. And this argument was the last ball for us that has turned the balance in favor of an agricultural project.

We believe that taking one protein in our iGEM project as an example we will demonstrate the potential of the technology for producing thermostable enzymes that can make livestock farming more effective both in Russia and in the world, which will eventually contribute to solving the global starvation problem as well.

Our product design is dictated by the product itself. Our project is aimed at creating a technology that will contribute to increasing biomass produced by farms and at the same time reducing these farms’ production expenditures.

Phosphorus is one of most important macronutrients vitally essential for effective bodily functions. Carnivorous animals receive it from meat and fish. The main source of phosphorus for herbivorous animals feeding mostly on plants are grains and beans, but most part of phosphorus here is bound in a substance known as phytin and its derivatives. Phytin – a small organic molecule derived from hexahydric alcohol – is a ‘depot’ of phosphorus in plants. In its natural condition phytin is not absorbed in animal’s body; for this purpose a particular enzyme, phytase, is required. All these factors should be taken into account when producing compound feeds for farm animals.

In order to improve nutritive properties of a final product various phytases are used as a feed supplement today. However, production of granulated feeds which are most convenient for consumers requires drying when feed components are exposed to quick heating up to 70°С. Therefore, enzymes that cannot survive such temperatures are not used in compound feeds. As a result, a wide range of enzymes turns out to be excluded from feed production. Moreover, the market of feed enzymes mostly provide enzymes that are poorly adapted to animals’ gastrointestinal tract (GIT) conditions. To solve this problem we suggest developing production of enzymes that are more efficient in these conditions.

For the purpose of enzyme mass accruement, we will use Yarrowia lipolytica cells serving as microcapsules for the enzyme. This will help us to increase our enzyme’s immunity to quick heating due to colloidal structure of yeast cell content. A good example of a working enzyme is phytase derived from Obesumbacterium proteus. It maintains thermal stability up to 60°C and is able to survive animals’ GIT conditions. But here we face another problem: it is prone to quick breakdown when accumulated in yeast cells. To stabilize phytase of C. braakii in our project we suggest to create a protein in which a small domain of Cellulose-binding domain, a protein with a stable tertiary structure, will be attached to N-end of phytase. This will serve as a barrier between phytase and proteolytic enzymes of yeasts.

Besides, it is a well-known fact that one of ingredients used for compound feed production, namely meat and bone meal, is subject to fungal contamination and about one third of raw materials becomes unusable for this reason. Our choice of this specific type of yeasts has not been occasional: these yeasts are able to consume toxins emitted by fungi. This makes it possible to salvage damaged raw materials, which is the last but not least argument in favor of our product design choice.

Though vegetarianism has being recently on the rise in many developed countries, meat still remains the main source of many essential nutrients for food consumers all over the world and livestock farming is gaining pace to meet the growing demands of the increasing world population. And this is one of the reasons for environmentalists to get more and more concerned.

Livestock production is generally known as one of the factors contributing greatly to such environmental problems as global warming, increasing energy consumption, acidification and aquatic eutrophication. But how can phytase help to reduce the impact?

Phosphorus is an essential nutrient for healthy development and normal bodily functions of farm animals and poultry. This mineral is naturally found in many feed ingredients but only a part of it is available to animals as it is bound in phytic acid. In order to solve this problem phosphorus is often added to feeds in the form of supplements. For this purpose inorganic phosphates are extracted from natural rocks. But such phosphates are limited and exhaustible resources which are used not only for feed production (for example, they are widely used for producing plant fertilizers). What is even more discouraging is that a great amount of this phosphorus added as a supplement is emitted to the environment with the manure leading to soil acidification and – along with nitrates and ammonia – eutrophication when it runs off into water systems and is accumulated in water reservoirs, thus contributing to algae bloom which can finally result in total aquatic fauna and flora extinction. There are tough requirements in some countries regulating the disposal of excessive manure. But are these requirements always accurately observed? And what about those countries which do not have such regulations? In the long run this means that we waste exhaustible natural phosphorus resources on exterminating life, which is absolutely illogical. The advantage of using phytase in feeds is obvious: it makes it possible to minimize the necessity for supplementary phosphorus as phosphorus can be easily derived from feed itself. The other side of the ‘phosphorus supplement production and manure disposal’ coin is energy consumption and contribution to global warming effect which could also be reduced by using phytase as its production is less energy consuming as some recent studies have proven and the manure excreted in this case contains less phosphorus and more of it can be applied to lands.

In order to verify our conclusions that had been mostly drawn from literature review we sought for an expert opinion of a person dealing with environmental challenges in a professional way. Natalia Shchegolkova (D.Sc. in Biology, a leading researcher at the Institute for Water Problems of the Russian Academy of Sciences) believes: “Scientific and technological solutions aimed at redistributing biogenic elements are extremely important as biogenic element cycles drive many energy problems and the problems of preserving favorable ecological environment, as biogeochemistry achievements have demonstrated. The project proposed here is one of such solutions. Producing and using phytase can make phosphate supplements to feeds more effective, reduce their amount and even eliminate the necessity to use them at all, which undoubtedly results in solving a number of ecological problems: reducing energy consumption in the system ‘environment – livestock – human’, improving the quality of manure, saving exhaustible recourses (mineral phosphorus) and – indirectly – solving the problem of water contamination (when using manure in agriculture instead of landfill depositing).”

Thus, using phytase in feeds contributes to eliminating or minimizing a number of negative factors of environmental impact in general and our improved phytase makes natural phosphorus absorption even more effective and the environmental impact even less depressing in particular.

We believe that our project is absolutely safe for humans as none of the microorganisms used (Yarrowia lipolytica, Escherichia coli, Obesumbacterium proteus) is pathogenic for people. When dealing with AMR factors we have applied all the required safety measures to prevent distribution of antibiotic resistant bacteria (through sterilizing space, devices, etc.). The result of the project is obtaining feeds from which more phosphorus will be absorbed in animals’ organisms. At first sight, it can be assumed that this can lead to accumulation of phosphorus in meat ant its excessive concentration in food. But these are only speculative concerns as we should remember about two conditions. First, currently non-organic phosphorus is added to compound feeds for better animal growth, which presents even more risks for consumers as this increases the risk of using low-quality phosphorus supplements containing accessory substances that can be harmful for live organisms. In this sense our project is rather a benefit from the viewpoint of final product safety: we create a technology ensuring consumption of phosphorus from natural materials. Second, even if the amount of phosphorus in feeds is excessive it is not absorbed in the organism and is removed with excrements. Therefore, concerns about the quality of final meat products are absolutely groundless.

The first question you have to answer when dealing with bioengineering and gene modification is if it’s legal or not. Since GMOs have become ingrained in our everyday lives, the society has split into GMO advocates and GMO opponents. And Russia is among those many countries where there is no consolidated opinion on this matter. But the government seems to be giving ear to GMO opponents more and more.

Gene engineering has been under government regulation in Russia since June 05, 1996 when the Law on Government Control over Gene Engineering Activities came into force. The Law mostly provided relevant definitions and specified terms for safe and secure gene engineering activities. It can be said that the Law was quite progressive and forward-looking as it did not set any obstacles to gene engineering in general but rather promoted the idea of its controlled safety and required disclosure of information about such safety.

But time passed by, the matter of gene engineering and GMOs was expanding beyond the walls of laboratories and research institutions capturing minds of more and more common people and ultimately evoking the mob GMO hysteria among information consumers. As one of the results of such hysteria, in 2014 over 100 thousand Russian Internet users signed a petition against GMOs addressed to the President. In the beginning of 2015, a draft law was submitted to the State Duma (the lower chamber of the Russian Parliament) prohibiting cultivation of genetically engineered plants and animals in Russia. On July 03, 2016 the law was signed by the President. It was Federal Law No. 358-FZ amending a number of previously adopted laws on gene engineering activities. Among other restrictions, the Law prohibits:

(Article 2): importing to the Russian Federation and planting seeds of such plants the genetic program of which has been modified using gene engineering methods and which contain gene engineered materials that cannon be the result of natural processes; the exclusion is planting such seeds for expert evaluation and research;

(Article 4): growing and breeding such plants and animals the genetic program of which has been modified using gene engineering methods and which contain gene engineered materials that cannon be the result of natural processes; the exclusion is the growing and breeding of such plants and animals for expert evaluation and research.

The law split the society even worse. The advocates of the Law still believe that the environmental and human safety of genetically engineered organisms has not been proven; their opponents emphasize that their harm has not been proven either. And the debate seems to be endless…

For the purpose of our research project, we use genetically modified yeasts which are neither plants nor animals; thus, we do not breach the Law. Besides, we believe that the banning of GMOs affects innovations in agriculture which is the matter of economic security. Therefore our team's position on the GMO and bioengineering problem is as follows: we believe that being young scientists with a more profound knowledge in the field of synthetic biology than a general consumer we could become a kind of translators of the idea of safety and reasonability of GMO food production and consumption and this way oppose the prejudice about GMOs harm in the future.