Sustainable Development Goals

Inspiration

Carbon neutrality is one of humanity's most important common issues. ONCE is also thinking about applying its power to help the cause of carbon neutrality for all humankind.

During the Nature Matters conference, we had an in-depth discussion with the government of Yancheng City about the development of synthetic biology in the era of carbon neutrality. The Yancheng city government officials discussed two environmental dilemmas they are facing.

They revealed to us a counterintuitive conclusion: In the current biosynthetic manufacturing scenario, due to factors such as production efficiency and maturity of the manufacturing process, biosynthetic products do not perform as well as their chemical counterparts in terms of carbon neutrality throughout their life cycle, leaving a larger carbon footprint in the entire value chain. The traditional petrol-based industry has contributed most of the chemicals in our daily lives. Microbial fermentation producing chemicals from a renewable resource is environmentally friendly and sustainable to replace the petrol-based industry. However, typical microbial fermentation often faces the problem of easy contamination and thus requires complicated sterilization and high energy input.

In addition, synthetic biology is produced at a much lower scale at this stage than in the traditional chemical industry. Small-scale biosynthesis requires higher energy consumption to produce a unit of product than chemical synthesis; therefore, the carbon footprint impact from energy consumption is very high.

Yancheng City is one of the first new energy demonstration cities in China. The scale of offshore wind power in Yancheng is 3.52 million kilowatts, accounting for 40% of China and 10% of the world. Yancheng is known as the "First City of Offshore Wind Power." To eliminate the original sin of carbon emissions caused by the limited production scale, some of the leading Chinese synthetic biology companies have chosen to use the "offshore wind power" technology. Some of China's leading synthetic biology companies have chosen to use wind power as a clean energy source to reduce carbon emissions from industrial production. But in reality, if we convert wind energy to coal energy, synthetic biology companies can do no more than traditional companies in terms of carbon neutrality.

In addition, it also consumes a large amount of freshwater for preparing the culture medium. The desalination industry in Yancheng is relatively developed, but traditional desalination is a highly energy-consuming industry, with more than 150 countries and regions worldwide building thermal power plants to provide the electricity needed for desalination. However, traditional desalination is a highly energy-consuming industry, and more than 150 countries and regions worldwide have built thermal power stations to provide the electricity needed for desalination.

The Yancheng government personnel asked us, "Can the strains you use be cultured directly using unsterilized seawater rather than using our freshwater resources derived from desalination, plus sodium chloride extracted from the seawater?"

At the same time, they also wanted us to try to culture V. natriegens with marine aquaculture wastewater, which contains mainly organic matter, ammonia nitrogen, nitrite, nitrate, and phosphorus. It is mainly characterized by the large volume of water, few types of pollutants, and small variations in content. The pollutants are mainly organic matter, nitrogen, phosphorus, and other nutrient salts, and we speculate that these characteristics will probably favor the rapid growth of V. natriegens. Identifying a suitable microbial strain, excluding expensive sterilization and freshwater consumption, is highly desirable for developing energy-efficient and sustainable biotechnology.

To address the above problems, Vibrio natriegens, a good microbial chassis with low nutritional requirements, high salt tolerance, and rapid growth rate, can be selected as the host for chemical production. This bacterium is non-pathogenic and belongs to biosafety level 1.

We are seeking to use non-sterilized fermentation of V. natriegens with seawater. To systematically verify the project's feasibility, we plan to seek the community's help to collect seawater from China iGEMer for non-sterilized fermentation.

Experimenting with a novel microchlorinated V. natriegens culture

V. natriegens requires sodium ions (Na+) for cell proliferation. The genome features genes for at least one Na+-extruding oxaloacetate decarboxylase, Na+-transporting NADH: ubiquinone oxidoreductase, and Na+-translocating ferredoxin: NAD+ oxidoreductase.

The sodium ion is necessary for the growth of Vibrio albicans. We designed a defined medium containing only traces of this halogen but sufficient sodium ions to maintain the high growth rate of V. natriegens. This cultivation method will greatly reduce the problem of chloride corrosion. Unfortunately, the laboratory or industrial cost of this kind of medium is higher than those of the medium directly added with sodium chloride. We must evaluate this new medium further and find a more economical and feasible industrial fermentation way.

Yancheng inspection: another way out for sodium chloride

Yancheng City is the only prefecture-level city in China named after "salt." Yancheng has a history of more than 2,000 years of sea salt development and is a city of sea salt that has been developed by sea salt culture in the true sense of the word. Yancheng is an important production base for sea salt, and its vast mudflats provide excellent conditions for "cooking sea salt." The culture conditions of V. natriegens are closely related to sodium chloride, and adding sodium chloride contradicts the large-scale industrial fermentation of V. natriegens. For this reason, we went on a field trip to Yancheng to find a way to coexist between sodium chloride addition and large-scale industrial biofermentation production.

A visit to the China Sea Salt Museum: A storage solution for highly concentrated sodium chloride

The history of humanity's interest in the sea can be traced back to an even longer time before the invention of "boiling the sea for salt." In the face of the surging sea and the vast expanse of the mud flats, the ancestors converged on the flat, open mud flats and used the mud flats and grass swings to create a chapter in the history of sea salt extraction. With the socio-economic development, manual salt production has receded from the historical stage, the modern salt production process and procedures have become much more efficient, and salt production has increased greatly. We imagine that if V. natriegens steel tank fermentation faces a high concentration of chloride ions corrosion, the whole process of industrial salt production, storage, and transportation will face more serious corrosion. A systematic understanding of salt's industrial production may help us solve this industry problem once and for all.

We visited the China Sea Salt Museum to see the historical changes in how salt is produced and to ask the museum staff our questions. The museum staff showed us the current FRP brine storage tanks used in industrial production. He highly recommended that we consider using Fiber reinforced plastics (FRP) for the fermentation production of V. natriegens. FRP is widely used in brine storage tanks and brine pipeline transportation in the sea salt industry. Fiber-reinforced plastics can withstand up to 30% salinity. It has excellent oxidation, acid, and alkali resistance and is resistant to aging. It is also cost-effective, environmentally friendly, and potential for industrial mass production applications.

The wisdom of the countryside: the inspiration of the biogas digester

Young scientists often work in academic institutions and laboratories, as does ONCE. Since ancient times, China has been known as a largely agricultural country, with 94.7% of the country's land area in rural areas, but ONCE members live and work in large cities, so while we design and complete projects based on real-world needs, we also hope to be inspired by the countryside, where we are rarely found.

During a field trip to Dongzeng Village in Binhai County, Yancheng City, we were surprised to find that Fiber reinforced plastics are already widely used for microbial fermentation in the Chinese countryside. Fiber-reinforced plastics are already being used on a large scale in constructing methane-generating pits. We were fortunate enough to witness the installation of an FRP digester in the village of Dongzeng. Through the interview with the installation engineer, we learned that Fiber reinforced plastics are widely used in the fermentation culture of microorganisms of biogas fermentation, and the whole process of biogas generation is as follows. 1. Hydrolysis stage: The fermentative bacteria use the extracellular enzymes secreted to decompose the organic matter in vitro enzymatically. 2. Acid production stage: The fermentative bacteria decompose the small molecule compounds produced in the hydrolysis stage into acetic acid, propionic acid, and butyric acid. Then hydrogen-producing acetic acid-producing bacteria convert the propionic acid and butyric acid produced by fermentative bacteria into acetic acid, hydrogen, and carbon dioxide.3, methanogenic stage: methanogenic bacteria convert hydrogen and carbon dioxide into methane, and the latter decarboxylates acetic acid to produce methane.

At the same time, we witnessed the installation of FRP microorganisms for biogas fermentation. We saw for ourselves the good overall performance of FRP fermenters, the ease of transportation and installation, the adaptability of installation, the ease of management, and the quickness of operation and construction. We were convinced to use FRP as the material for V. natriegens. industrial fermenter material. Of course, during the follow-up communication with the industry, we also learned that FRP fermenters might have defects such as the inability to install high-powered stirring devices, weaker processing performance than stainless steel, easy-to-shed impurities, etc. In the follow-up small trial and pilot scale stage, we decided to further study and implement the use of Fiber-reinforced plastics for V. natriegens according to its biological characteristics. The idea of using Fiber reinforced plastics material for V. natriegens production fermentation was further investigated in the subsequent small-scale and pilot scale-up stage.

Non-sterilized fermentation and natural seawater culture

After the trip to Yancheng, We discussed the possibility of Non-sterilized fermentation and natural seawater culture. The traditional microbial fermentation process requires high-temperature sterilization of media and fermentation equipment, and the requirements for operators, fermentation conditions, and fermentation procedures are very strict. If the equipment or media are not thoroughly sterilized, the seeds carry undesirable microorganisms during inoculation, or the fermentation equipment is poorly sealed, microbial contamination will arise. Microbial contamination affects the quality and safety of biological products and causes significant economic losses. This problem limits the development of technologies for producing chemicals through conventional fermentation and puts it at a disadvantage in competition with the traditional petroleum-based chemical industry.

Non-sterile open fermentation, a method of producing strains using specific growth pressures without sterilization of fermentation equipment and media, is considered a promising next-generation industrial biotechnology due to the simplicity of the fermentation process and energy cost savings. After 65 days of continuous open fermentation, the cell dry weight of the recombinant strain reached 65 g/L, and the PHB content was 70% of the cell dry weight.

In recent years, global freshwater reserves have been decreasing rapidly, but the demand for domestic and industrial water is increasing, and the problem of freshwater scarcity is becoming more serious. The traditional industrial microbial fermentation process requires many freshwater resources, which will undoubtedly aggravate the problem of freshwater scarcity. Seawater is the most abundant water resource in the world, and the development of a non-sterilized fermentation process using seawater resources instead of freshwater is of great significance to save energy and water resources and achieve the goal of green and sustainable development.

Composition of seawater

Seawater is a complex solution in which the components with content greater than 1 mg/kg are the main components of seawater, in addition to the hydrogen and oxygen that make up the water molecules and non-conservative dissolved silicic acid, a total of 11 species. The six components with the highest content are, in descending order, chlorine, sodium, sulfate, magnesium, calcium, and potassium, accounting for 99% of the total salt content of seawater.

Dissolved oxygen is an indispensable substance in marine life activities, mainly from the atmosphere and phytoplankton photosynthesis. The amount of dissolved oxygen in water is related to atmospheric pressure, water temperature, and salinity.

Here we focus mainly on the surface layer (because the place to take water can only be surface water). Wind, waves, and vertical convection stir the surface layer, and oxygen exchange between surface water and the atmosphere tends to be balanced more quickly. The dissolved oxygen in the surface water is basically at saturation, a condition saturated with oxygen, and will probably be favorable for V. natriegens cultures with very high growth rates.

The salinity of seawater is a scale of the salinity in seawater and is one of seawater's most important physicochemical properties. The absolute salinity of seawater is the ratio of all dissolved solids in seawater to the weight of seawater, usually expressed in grams of salt per kilogram of seawater. The average salinity of seawater is about 35.

V. natriegens culture conditions

ATCC14048 is the most used strain of V. natriegens at present. The PH required for its growth: is 6-9.5, with an optimum of 7. ATCC14048 grows differently in different salinities. The average PH of natural seawater is stable between 7.9-8.4, and the average salinity is about 35, although the salinity of near-layer seawater is unevenly varied. However, we used reasonable assumptions based on the recommended culture conditions for V. natriegens: using natural seawater or aquaculture wastewater with additional carbon sources to make the medium could better meet the growth of V. natriegens. In the subsequent experiments, we verified the conclusion and conducted further investigation and analysis.

Summary of Sample Information

L'Oreal Interview

L'Oreal is one of the world's leading beauty and personal care brands, with 35 brands and a presence in 150 countries, bringing the latest technological advances in beauty and personal care products. Recommendations. During the Skyworks Bioeconomy Summit, we had an in-depth conversation with Rebecca Xu, Head of Open Innovation at L'Oreal's China R&D and Innovation Center, and Sarah sun, Head of Ingredients. There we discussed the value of our ideas to sustainability

"L'Oreal is committed to transforming its business model to respect the planet's boundaries. In the context of carbon neutrality, consumers' expectations of the beauty industry are moving towards "sustainable beauty." Based on the Earth Boundary Theory. Based on the Earth Boundary Theory, the company focuses on four areas in setting sustainability goals: climate protection, water management, biodiversity, and recycling of resources. In response to the strategic goal of "L'Oreal for Tomorrow," L'Oreal's research and development department has developed a green science concept for its ingredients and formulations. On the one hand, they will evaluate the environmental footprint of existing technologies to ensure that they respect the planet's boundaries; on the other hand, they will actively promote technological upgrades and transformations to face the growing environmental and social challenges.

Alpha-Bisabolol is available in synthetic and natural sources. Synthetic alpha-Bisabolol contains only 46% of the active levorotatory isomers. Natural alpha-Bisabolol is generally extracted from the Brazilian Candia tree. Most manufacturers on the market today, including L'Oreal, extract alpha-Bisabolol from the essential oil of the Brazilian Candeia Tree (Eremanthus erythropappus).

In recent years, the Amazon rainforest, known as the "lungs of the earth," has become increasingly difficult to "breathe," and deforestation is a key cause of forest fires in the Amazon. Deforestation is a key cause of forest fires in the Amazon. The area damaged has reached its highest level in the past 15 years. In contrast, Candeia Tree's alpha-Bisabolol production is very low, with only 7 kg of alpha-Bisabolol extracted from 1 ton of bark, with a 10-year planting period per tree and official harvesting restrictions, Candeia Tree is highly non-renewable. The addition of alpha-Bisabolol to L'Oreal's star products, the Hair Serum and the Cleansing Deep Cleansing Oil, is not in line with the "L'Oreal for Tomorrow - 2030 Sustainable Development " goal. Two L'Oreal leaders praised ONCE's initiative to use a synthetic biology platform to replace traditional destructive plant extracts. They believe this initiative is very much in line with L'Oreal's values. They expressed great interest in inviting us to become one of L'Oreal's raw material suppliers once we achieve mass production of alpha-Bisabolol, which could help L'Oreal further implement reaching sustainable beauty.

Distributors Interview: High Quality, Mass production and On-Time Delivery rate are the primary factor when they make cooperation.

We discussed with many distributors the opinions on our sustainable development goals. Distributors tell us that traditional bisabolol production requires a lot of tree felling, and the chemical synthesis method has a large purity problem. Therefore, if it is necessary to sell in the country, quality, scale-up production and timely delivery will become the main issues. We listened to them and put them on our to-do list

Promoting on social media and Diving activites

We started by contacting some of our diving buddies in Bali to ask them about their views on the ocean, skincare and recycling. And with them, experience underwater diving near Penida Island to cultivate recyclable and technological skincare concepts.

A ONCE company was founded- A LEADING SYNTHETIC BIOLOGY APPLICATIONS PLATFORM

Based on all the trials before, we decided to start a company to achieve our sustainable development goals. ONCE_CHINA is a company focused on synthetic biology beauty and food upstream, committed to using synthetic biology application technology to achieve Carbon neutrality and transformation. The company's core technology is synthetic biology application product manufacturing technology. As a research-oriented, customer-centric enterprise, the company provides customized raw material solutions for downstream enterprises such as beauty and food in a sustainable way. The company pioneered the use of Vibrio sodiums as a chassis organism and derived several pipelines on this basis.

We had communicated with Sequoia China、Heli and many other Ventures shows Great interest in ONCE, and we had communication with local government (ZHANGJIAGANG, JIANGSU, CHINA)