The main goal of the iGEM NU Kazakhstan team 2018 was not only to create the cyanobacteria which can transform toxic hydrogen sulfide compound to elemental sulfur and hydrogen gas in laboratory conditions but also to investigate how to integrate this project in the real industrial situations.

Kazakhstan is commonly known to be the major producer of oil in the world; it has an enormous amount of oil resources: 30 billions of barrels, 172 oilfields, 1.8% of global oil reserves [1]. The figures speak for themselves, and Kazakhstan tries to fully use its potential. However, the major problem of an oil refinery which challenges not only our country but also other global oil exporters is the presence of hydrogen sulfide in crude oil. Decayed organic matter (kerogen), which is trapped in sedimentary rocks, serves as the main reagent of thermal conversion, in which the oil is formed as a major product. Kerogen, which is highly concentrated with sulfur, releases hydrogen sulfide during the reaction and this gas stays in crude oil.

Hydrogen sulfide is highly toxic gas, which in high concentrations can interfere with the organism’s capability of cellular respiration. The human body can block its effect by transforming it into sulfate, using hemoglobin-bound enzymes of blood or liver cells. However, when the concentration of hydrogen sulfide is too high, the organism’s ability of detoxification is overwhelmed and it eventually leads to death. Besides these health risks, the presence of H₂S in the oil can lead to the corrosion of pipes and it is of the high cost to replace them periodically.

Our team decided to know more about the process of hydrogen sulfide purification and details related to the integration of the project by contacting with the variety of industrial companies in Astana, Atyrau and Almaty. Our team representatives met with one of the largest oil company directors in Atyrau, as they are located there and most of Kazakhstan’s oil reserves are from the Atyrau region. Below you can see the list of meetings and some outcomes of these meetings.

NCOC is the main offshore oil and gas developer in Kazakhstan. Their estimated oil production for 2018 is 370,000 barrels per day, which is more than their expected 75,000 barrels per day in 2016 [1]. They have five main fields on the Caspian Sea about 80km from Atyrau city. NCOC faces many safety, logistical and engineering obstacles due to its location. The oil from these fields has high hydrogen sulfide content as well as carbon dioxide. The principle system used for the sour gas treatment is the raw gas re-injection. Their shareholders (KazMunayGas, Eni, ExxonMobil, Royal Dutch Shell, Total S.A., China National Petroleum Corporation, Inpex) have been practicing raw gas re-injection in many different environments. This technique has been invented in Canada in 1989 and practiced in 39 projects in the world, including the Karachaganak and Tengiz fields in Kazakhstan. With this process, 80% of sour gas from crude oil is returned to the reservoir, while the rest 20% is generated to produce elemental sulfur. They estimated that 3800 tonnes of sulfur per day will be produced in their project [1].

Our team representatives visited the NCOC head office in Atyrau. During the meeting, we introduced them to iGEM, synthetic biology and we explained our project. They were very responsive as they were interested in our new approach to the bioremediation of sour crude oil wastewater. Even though they were hesitant about the risks of genetically modified organisms, they assured that it has good potential to turn into a commercialized project that they can apply in their wastewater treatments.

Firstly, they were interested in a specific design where our cyanobacteria can carry out the bioremediation of sour crude oil wastewater. We approached this comment by offering an approach that includes keeping the cyanobacteria in a gel-like reservoir, where the wastewater can be pumped in.

Secondly, they were concerned with what will happen to cyanobacteria after it has taken up hydrogen sulfide from the wastewater. We replied that as part of our risk management, we will be using the SuperNova, a variant of KillerRed, which uses Chromophore Assisted Light Inactivation that will lead to self-destruction of the cells at specific wavelengths of light.

For companies like NCOC, the elemental sulfur produced after the treatment of hydrogen sulfide is sold to overseas companies. We proposed a different solution. NCOC re-injects 80% of sour gas back into the reservoir. If this is treated with cyanobacteria and they are burned after they have carried out hydrogen sulfide reduction, then these can be turned into nanomaterials. The acid gas (hydrogen sulfide with carbon dioxide) re-injected back into the reservoir poses serious environmental problems in the future, as the widespread release of hydrogen sulfide due to the leakage of wellbores is a possibility to be considered [2]. According to NCOC’s report, their air quality monitoring stations registered a number of instances where the hydrogen sulfide concentrations exceeded the Maximum Permissible Concentrations in the Atyrau region due to leaks [3].

NCOC’s real-life applications questions helped to improve our project because many aspects of the project were discussed as if it was going to be applied in their production processes. This led us to examine the parts of the project that needed to be well thought out not only for laboratory conditions but also how it could be practiced in mass production at different conditions. Overall, this meeting with NCOC gave us valuable feedback and they look forward to collaborating with us, as they invited us to intern at their laboratories where they have all the samples we need to further assay the viability of our cyanobacteria. We also look forward to continuing this project with NCOC after the Giant Jamboree, because the possibility to decrease the environmental risks of hydrogen sulfide in our country is our priority.

In 1922, the company was developed for geological exploration, development of oil and gas fields as well as oil and gas production and treatment. The company exports oil along with selling it in the domestic market. Their produced gas is sold through the national gas supplying company. In June 2017, EMG launched a project for a treatment unit that will produce commodity gas and commercial granular sulfur [4]. EMG uses the LO-CAT Merichem technology for the production of elemental sulfur. LO-CAT Merichem is a system that converts hydrogen sulfide to elemental sulfur with the use of a chelated iron in special containers and it guarantees hydrogen sulfide removal [5]. EMG allocates funds for projects to clean historically contaminated areas for the oil sludge treatments. They use the Finnish Kalottikone Unit for the soil recycling that is later used in construction and repair of roads. The Kalottikone Unit is a thermal desorption equipment that cleans contaminated soil from oil products [6].

We organized a meeting with the executive directors of EMG in Atyrau. We got to introduce them to iGEM and explain our project. After a short presentation, we discussed with them the possibility of applying our project in their processes. They were positive about the project and offered their help for the expansion of the project in real life. However, they were a little sceptical about the genetically modified organism that would be used to treat the sour crude oil wastewater. We assured them about the safety measures we have thought about, which eased their worries about the risks of the project.

They explained the operations of their company. As their company extracts the oil from the ground, they separate the extraction into oil, gas and water. The sulfur content of the oil meets the minimum requirements and the standards of the legislature of RoK; therefore, they export or sell the oil. The extracted gas is then further produced as commodity gas that is sold for local usage. The water is pumped back into the reservoir to create the proper pressure for the oil to be pumped closer to the well in a process called water flooding [7].

Since our project is concerned about the wastewater, we tried to show that water flooding is not a reliable process even if it can increase the recovery factor and the production rate of oil extraction. We assume that their technologies (LO-CAT and Kalottikone) would not reduce the environmental factors as much as our synthetic biology approach would. Firstly, in our approach, the hydrogen sulfide reduction yields hydrogen gas and elemental sulfur, which are both potential biofuels. Secondly, the wastewater that is pumped back to create pressure can be treated with our cyanobacteria, because our process does not reduce the volume of the wastewater. Therefore, it would be producing the favorable byproducts, while creating the appropriate pressure. This solution was something new for the executive directors, but they were open to new processes.

The meeting with EMG did not only give us beneficial feedback, but they helped to clarify some oil production processes that we could not find online. They are ready to give their wastewater samples to analyze the potential of our genetically modified cyanobacteria. We look forward to collaborating with EMG, as our project can be applied as part of their oil and gas treatment processes.

The Dow Chemical Company (based in Midland, Michigan, USA) is one of the largest chemical manufacturers in the world. This company is involved mainly in the manufacture of plastics, chemicals and agricultural products. As the hydrogen gas is the byproduct of not only oil refinery reactions, but also different industrial processes such as paper, rayon and leather manufacture, we decided that the opinion of people specialized in chemicals production is also very important. We organized the meeting in Astana city with the representatives of this company. During this meeting, many significant conclusions were drawn:

1. The process of hydrogen sulfide conversion can be used not only in oil & gas refineries but also for sewage treatment. There is the serious problem of hydrogen sulfide remained after water treatment, which can be recognized through the strong smell present in some streets in the city of Astana. The country manager of this company also underlined that our project would find easy application namely in this case.

2. The use of cyanobacteria in hydrogen sulfide conversion would be really challenging in oil industry application. There are two reasons for this statement: light delivery and speed of flow. Under the conditions of factories, it is unknown how possible will it be to provide light for maintaining the life cycle of cyanos and what amount per hour they can really deal with.

Overall, the experts were really inspired by our project and they continue to support us by providing with essential contacts and useful advice. With their help, we contacted the ChemInvest company which promised to provide us with the oily water samples

1. To be realistic. The project itself is very interesting and it would be wonderful to apply such advanced principles of synthetic biology in one of the most challenging problems of the oil industry, but there are many unanswered questions such as the possible ways for storage of hydrogen gas, the capacity of H₂S consumption by cyanobacteria, the disposal of cells excess and other details. They tried to explain to us that we should think more practically and consider many unexpected issues.

2. To know how to estimate the cost of the project. The company representatives told that economical aspect is essential for the project of any type and in order to integrate the project successfully, you need to know the cost of every detail participating in the process. In addition to this, they mentioned the significant price of the Claus process currently used in the desulfurization of oil and stated that finding the cheap alternative to this method would drastically decrease their expenses.

3. To know how to work with information. They strongly recommended us to search more for the information related to the amount of hydrogen sulfide found on the fields, to accurately apply the statistical methods for analyzing this data and to present it in an appropriate way.

Such meetings always play a crucial role in creating and designing projects, because it has shown us that many aspects should be considered. Also, they agreed to provide us with their samples of oily water from the fields and to coordinate us throughout the project.