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From Science & Research

From Science & Research Enzymes to boost plastic sustainability The power of evolved ancestral enzymes and biotechnology Plastic pollution has become one of the most pressing environmental issues, as the rapidly increasing production of disposable plastic products overwhelms the world’s ability to deal with them. Global plastic waste generation more than doubled from 2000 to 2019 to 353 million tonnes. Nearly two-thirds of plastic waste comes from plastics with lifetimes of under five years, with 40 % coming from packaging, 12 % from consumer goods and 11 % from clothing and textiles. To repair the present, building and implementing a more circular economy is key to sustainable growth and addressing challenges like climate change, with targeted measures at all levels of society to address the challenges posed by plastic pollution. The circular economy also needs optimal waste management to promote plastic waste efficient recovery and recycling to avoid further plastic littering, eventually contaminating our soils and seas. Initiating the “RevoluZion” As a holistic solution to plastic pollution, the participants of the RevoluZion project are developing innovative biobased formulations, combining advanced enzyme engineering techniques together with biodegradable plastic resins as a matrix with programmed biodegradation to conciliate ondemand biodegradation in different managed (industrial and home composting) and unmanaged environments (soil, freshwater, marine water). The idea of the project is to reduce the typical times for composting to fasten the industrial treatment of compostable plastic articles and make it attractive and economically viable, or to allow compostability in domestic conditions and/or biodegradability in open environments (water, soil) of resins exclusively reserved for industrial composting. The formulations could serve different applications, for example, to produce coffee capsules or articles for agriculture. Therefore, the obtained blends of polyesters as matrix and enzymatic functional additives aspire to contribute as an integral solution to plastics waste management. In order to design highly active and robust enzymes for programmed biodegradation and compostability, RevoLuzion project is bringing together cutting-edge protein engineering methods based on directed evolution strategies and ancestral resurrection. The transformation processes of plastics through extrusion and compounding processes usually involve high temperatures and shear, which can negatively affect the activity of the enzymes developed. A delicate process of encapsulation and integration of such enzymes will be carried out to inhibit any associated degradation from the enzyme structures during its integration into the thermoplastic matrix. The aim of the project is to develop up to three prototypes of innovative biobased bioplastic materials using the disclosed advanced enzyme technology for different sectors of application of the plastic industry: • Food packaging: to enable home compostability for thick articles like meat trays and other containers. • Coffee capsules/pods: to reduce typical industrial/home composting time down to a third without impairing shelf-life capacity. • Agricultural films: to support excellent mechanical properties aboveground and biodegradation in soil without leaving residues. Grégory Coué Technical Manager Kompuestos Palau Solità i Plegamans, Spain The RevoluZion project is part of the Next Generation EU European Plan. Grant PLEC2021-008188 funded by MCIN/ AEI/ 10.13039/501100011033 and by the “European Union NextGenerationEU/PRTR”. “The consortium is made up of Kompuestos as the project leader, together with different topquality research centres and universities: AITIIP Tecnological Centre, the University of Granada and 2 research groups from the Spanish National Research Council (CSIC: CIB and ICP)”For more information about the project: AITIIP: CSIC-CIB: CSIC-ICP: Kompuestos: University of Granada: 22 bioplastics MAGAZINE [06/22] Vol. 17

Steel mill gases transformed into bioplastic Recently, a Korea-Spain joint research team recreated bioplastic from wasted by-products from gas fermentation from steel mills. Through joint research with Spain’s Centre for Research in Agricultural Genomics (CRAG – Barcelona), a research team led by Gyoo Yeol Jung, Dae-yeol Ye, Jo Hyun Moon, and Myung Hyun Noh in the Department of Chemical Engineering at POSTECH (Pohang, South Korea) has developed a technology to generate artificial enzymes from E. coli. The joint research then succeeded in mass-producing itaconic acid, a source material for bioplastic, from acetic acid in E. coli. enzyme can be used in E. coli to produce itaconic acid. With this technology, it is now possible to build a microbial cell factory that can easily produce itaconic acid from cheap and various raw materials. From Science & Research Recognized for its significance, this study was recently published in the Editor’s Highlights section of the international academic journal Nature Communications [1]. Comparison of itaconic acid production in the natural metabolic pathway in E. coli and the construction of a new itaconic acid biosynthesis pathway through the introduction of a new artificial enzyme. The itaconic acid production increased as a result. (Picture: POSTECH) Itaconic acid produced by fungi with membrane-enclosed organelles is used as a raw material for various plastics, as well as cosmetics and antibacterial agents. Although its global market value is estimated high at around 130 billion KRW (USD 91 million) this year, its production and utilization have been limited due to the complex production process and high cost of production. For this reason, studies are being actively conducted to produce itaconic acid with industrial microorganisms such as E. coli. Although E. coli can be produced using inexpensive raw materials and is easy to culture, additional raw materials or processes were required to produce itaconic acid since it lacks membrane-enclosed organelles. This research result is evaluated as a key original technology for producing itaconic acid from byproducts of gas fermentation products from steel mills, seaweed, as well as agricultural and fishery byproducts such as lignocellulosic biomass. By replacing the raw material from petrochemicals with biosynthesized itaconic acid, the new technology is anticipated to contribute to a carbon-neutral society and significantly expand the itaconic acid market. This study was supported by the C1 Gas Refinery R&D Program, the Mid-career Researcher Program, and the Basic Science Program of the National Research Foundation of Korea. AT Using biosynthesis, the joint research team developed an artificial enzyme to pave the way for E. coli to directly produce itaconic acid without membrane-enclosed organelles. The research results showed that the newly developed [1] Dae-yeol Ye et al, Kinetic compartmentalization by unnatural reaction for itaconate production, Nature Communications (2022). bioplastics MAGAZINE [06/22] Vol. 17 23

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