“A multi-enzymatic approach for PE-PFA mixed plastic waste biodegradation”
Mitigating the negative effects of single-use plastic consumption led to an increased usage of paper-based alternatives and even a complete ban of such consumption. However, this seemingly innocuous solution of using paper-based alternatives is still a burden to the limited capacities of landfills.
While planning the specifics of the project, one of our team members was inspired by an event hosted by Cebu's environmental enthusiasts in 2018. To her, one of the highlights was the discussion of unobvious non-recyclables such as paper cups. Unlike chip bags and tetra packs, the multi-material lamination of paper-based alternatives is unnoticeable; the plastic coating is inconspicuous to the naked eye. This led the team to explore other similar types of waste such as paper bubble envelopes, parchment/baking paper, paper food boxes/bags and glossy paper.
In Hong Kong, although the waste segregation system has improved to include more specific waste such as tetra packs, there is still no specialised waste processing system for broader paper-based alternatives, which have a popular demand in the local food and packaging industry. Motivated by this research gap, our team wants to investigate whether a multi-enzymatic approach can be developed into a technology to remediate this problem.
Over the last decade, the trend for sustainability has led to a considerable increase in the consumption of paper-based alternatives such as paper cups, straws,
milk cartons and other packaging materials (Foteinis, 2020). Despite the conventional wisdom that these products are recyclable, as they contain lamination
of additional materials, such as polyethylene (PE), per- and polyfluoroalkyl substances (PFAs) and aluminum (refer to Table 1), they are in fact not recyclable. These additional layers are source of microplastic
that affect the efficiency and the machinery of traditional paper mills and the value of the paper pulp (Yuhui, 2018). Hence, they are typically disposed of into the landfill or incinerated,
which are non-sustainable.
In the landfill, estimates have shown that paper cups take 20 years to decompose, with the possibility of microplastics in the leachate (Triantafillopoulos & Koukoulas, 2020). Additionally,
considering paper cups alone, Yuhui (2018) has estimated Mainland China distributes 10 billion cups annually. Australia, UK, and Taiwan, respectively, consume 2.5 billion, 2.52 billion, and 1.5 billion
cups per year (Foteinis, 2020; Yuhui, 2018). Moreover, there are 400 million plastic and paper cups being consumed every year in Hong Kong (SCMP, 2022). With this amount of waste generated and lack of
specialized waste processing facilities, using these products, thus, doesn’t alleviate the burden of landfills.
Paper-based alternatives | Coating |
---|---|
Paper Cups | PE |
Paper Straws | PE, PFAs or acrylic resin |
Paper Plate | petroleum plastic |
Paper food boxes/bags | PFA or PE film and coating |
In the above table, it is observed that the typical coatings are PE and PFAs. Both are known for their resistance
to degradation due to the chemical stability (Emblem, 2012; Krafft & Riess, 2015). Apart from chemical stability,
PE is also known for its low cost, durability, chemical inertness, and flexibility, which makes them highly suitable
and a popular choice for packaging materials (Emblem, 2012). In 2019, PE had a demand of 100 million metric tons,
which accounts 63% of the “global plastic resins demand”(Chitalia, 2020).
PE can be either mechanically or chemically recycled. Mechanical recycling involves remolding of old PE products
to form new products or pellets (Schyns & Shaver, 2020). The obvious disadvantage to this method is that it isn’t
sustainable. The properties of continuously recycled PE will be degraded over time until it must be completely
disposed of (Schyns & Shaver, 2020). It also can’t handle mixed and contaminated plastic waste streams (Vollmer et al., 2020).
In contrast, chemical recycling can upcycle diverse plastic waste into various fuels, monomers/oligomers (ready to be polymerized again)
through pyrolysis, solvolysis, dissolution, etc. (Vollmer et al., 2020). However, their recycling rates of these technologies must still
be improved (Vollmer et al., 2020).
On the other hand, PFAs are common in food packaging due to their heat resistance and water-oil repellency properties
(Krafft & Riess, 2015). Hence, due to its usage, it is estimated that in the last six decades, more than 47, 000 tons were
released from consumer products into the environment (‘About PFAs,’ n.d.). This is concerning as PFAs can bioaccumulate in the food
chain, consequently affecting human health, particularly on development and reproduction(George et al., 2023). Apart from bioaccumulation,
PFAs residues can directly migrate on food and drinks (Carnero et al., 2021). In America alone, Centers for Disease Control and Prevention's National
Health and Nutrition Examination Survey (NHANES) found PFAs in the blood of 97% of Americans, resulting in reduced blood levels of perfluorooctane sulfonic acid
(PFOS) and perfluorooctanoic acid (PFOA) (U.S. Department of Health and Human Services, n.d.). Despite these risks, the current proposed disposal is through chemical
breakdown in high temperature or incineration, which is not sustainable as it can release some of the compounds through the vapor (Verma et al., 2023).
Although the paper-plastic laminated waste recycling methods require sophisticated technology such as the ones mentioned above,
in Hong Kong, Mil Mill uses a physical approach where it separates the different layers: paper fibres, aluminium and plastic.
The paper pulp can then be upcycled by turning it into tissues, toilet paper, shoe boxes, etc. However, the remaining waste is mixed,
containing fibres, plastics and metals. Additionally, in the process of separation, plastics are physically processed into smaller
fragments. In this context, the method to process such waste can still be refined by lowering the cost, increasing the amount of
sample processed, and elimination of microplastics.
Compared to the approach of Mil Mill, biodegradation of this waste through different microorganisms is far more attractive, as
it does not only generate microplastics but also is a cheaper alternative. Generally, the mechanism of such biodegradation involves
the secretion of enzymes that could adhere to the surface of the polymer. The enzymes can then cleave the polymer to low molecular weight compounds.
These can then be taken up by the microorganisms for further reduction into water and carbon dioxide (Zeenat et al., 2021). In the context of paper-based alternatives,
Arumugam et. al (2018), have investigated the use of vermicomposting to biodegrade paper cups. They have proved that with the addition of a multiple bacterium,
it reduced the degradation time to 3 months and yielded a distinct separation of polyethylene on top of the compost.
With that, our solution involves the use of enzymes generated, typically by white rot fungi.
These are lignin peroxidase (LiP), manganese peroxidase (MnP), and laccase. These enzymes have
been proven to degrade PE and PFAs but are capable of degrading lignocellulose waste, such as paper.
Specifically, our team wants to first investigate the feasibility of using E.coli as a chassis
to secrete these fungal enzymes. To increase the enzyme yield in E.coli, codon optimization and
various RBS-ATG spacers were investigated. Three different fluorescent proteins were also used as
reporters to identify successful expression of the signal peptide secretion through pelB tag.
Using previous iGEM biobricks and their enzyme concentrations, we can then compare the efficiency
of our constructed biobricks.
After determining the optimal plasmid, the plastic degradation efficiency and enzymatic activity
will be determined. If a multiplex treatment is determined to be the optimum treatment, a multi-enzymatic
model will help predict the optimal ratio to degrade such mixed PE-PFA plastic waste.
After confirming the efficiency of the enzymes on pure plastic waste, the next step is to confirm their feasibility on degrading paper. Additionally, testing and developing the bioreactor and creating alginate beads that can immobilise the enzymes will be a future step that can get this project closer to its vision. This project, in the future, aims to offer a viable strategy for degrading single-use mixed plastic waste in a leaner, sustainable and environmentally friendly way without further sorting plastics into different types. Furthermore, our project hopes to create or kickstart the development of a system that accounts for the heterogeneity in plastic waste, particularly in the “bio-alternatives” of former plastic products (such as paper cups, paper plates, plastic-lined paper bags) for commercial use. This makes recycling easier. Besides, our findings can significantly benefit and elicit more interest from the recycling industry and future researchers in the potentiality of microbial enzyme degradation on mixed plastic waste and its incorporation in the commercial setting.
About PFAs. STEEP. (n.d.). https://web.uri.edu/steep/pfas/
Arumugam, K., Renganathan, S., Babalola, O. O., & Muthunarayanan, V. (2018). Investigation on paper cup waste degradation by bacterial consortium and Eudrillus eugeinea through vermicomposting. Waste Management, 74, 185-193. https://doi.org/https://doi.org/10.1016/j.wasman.2017.11.009
Carnero et al. (2021, June). Presence of Perfluoroalkyl and Polyfluoroalkyl Substances (PFAS) In Food Contact Materials (FCM) and Its Migration to
Food. Research Gate. https://www.researchgate.net/publication/352669399_Presence_of_Perfluoroalkyl_and_Polyfluoroalkyl_Substances_PFAS_In_Food_Contact_Materials_FCM_and_Its_Migration_to_Food
Chitalia, A. (2020). Why polyolefins are the polymers to watch. Retrieved Sept. 29 from https://www.woodmac.com/news/opinion/why-polyolefins-are-the-polymers-to-watch/
Emblem, A. (2012). 13 - Plastics properties for packaging materials. In A. Emblem & H. Emblem (Eds.), Packaging Technology (pp. 287-309). Woodhead Publishing. https://doi.org/https://doi.org/10.1533/9780857095701.2.287
Foteinis, S. (2020). How small daily choices play a huge role in climate change: The disposable paper cup environmental bane. Journal of Cleaner Production, 255, 120294. https://doi.org/https://doi.org/10.1016/j.jclepro.2020.120294
George, S. E., Baker, T. R., & Baker, B. B. (2023). Nonlethal detection of PFAS bioaccumulation and biomagnification within fishes in an urban- and wastewater-dominant Great Lakes watershed. Environmental Pollution, 321, 121123. https://doi.org/https://doi.org/10.1016/j.envpol.2023.121123
Krafft, M. P., & Riess, J. G. (2015). Selected physicochemical aspects of poly- and perfluoroalkylated substances relevant to performance, environment and sustainability—Part one. Chemosphere, 129, 4-19. https://doi.org/https://doi.org/10.1016/j.chemosphere.2014.08.039
Schyns, Z. O., & Shaver, M. P. (2020). Mechanical recycling of Packaging Plastics: A Review. Macromolecular Rapid Communications, 42(3). https://doi.org/10.1002/marc.202000415
SCMP. (2022). Hong Kong sends 400 million disposable coffee cups to landfills every year, according to green group. Young Post. https://www.scmp.com/yp/discover/news/environment/article/3185779/hong-kong-sends-400-million-disposable-coffee-cups
Triantafillopoulos, N., & Koukoulas, A. A. (2020). The Future of Single-use Paper Coffee Cups: Current Progress and Outlook. BioResources, 15(3), 7260-7287. https://bioresources.cnr.ncsu.edu/wp-content/uploads/2020/06/BioRes_15_3_7260_Review_Triantafillopoulos_Koukoulas_Future_Single_Use_Paper_Coffee_Cups_17365.pdf
U.S. Department of Health and Human Services. (n.d.). Perfluoroalkyl and Polyfluoroalkyl Substances (PFAS). National Institute of Environmental Health Sciences. https://www.niehs.nih.gov/health/topics/agents/pfc/index.cfm
Verma, S., Lee, T., Sahle-Demessie, E., Ateia, M., & Nadagouda, M. N. (2023). Recent advances on PFAS degradation via thermal and nonthermal methods. Chemical Engineering Journal Advances, 13, 100421. https://doi.org/https://doi.org/10.1016/j.ceja.2022.100421
Vollmer, I., Jenks, M. J., Roelands, M. C., White, R. J., van Harmelen, T., de Wild, P., van der Laan, G. P., Meirer, F., Keurentjes, J. T., & Weckhuysen, B. M. (2020). Beyond Mechanical Recycling: Giving new life to plastic waste. Angewandte Chemie International Edition, 59(36), 15402–15423. https://doi.org/10.1002/anie.201915651
Yuhui, M. (2018). Problems and Resolutions in Dealing with Waste Disposable Paper Cups. Science Progress, 101(1), 1-7. https://doi.org/10.3184/003685017X15129981721365
Zeenat, Elahi, A., Bukhari, D. A., Shamim, S., & Rehman, A. (2021). Plastics degradation by microbes: A sustainable approach. Journal of King Saud University - Science, 33(6), 101538. https://doi.org/https://doi.org/10.1016/j.jksus.2021.101538