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PET Hydrolase

One of the world's current biggest problems is the recycling of plastic. The stability of the polymers, their crystallinity and their hydrophilic surface make recycling difficult. Polyethylene terephthalate (PET) is one of the most widely used plastics today (around 30 million tons per year) and its recycling is now possible thanks to PET hydrolase, an enzyme isolated from the bacteria Ideonella sakaiensis ^[1]

Function

PET hydrolase (PETase) is part of a class of enzymes called esterases. These enzymes are hydrolysing enzymes capable of cleaving esters into an acid and an alcohol, with the help of water ^[2][3]. This enzyme is able to catalyse the hydrolisis of PET. Naturally, without enzymes, the degradation of PET can take hundreds of years. However, by using PETase, this degradation can be done in a mater of days ^[4][5]. Some scientists believe that PET degrading bacteria have developped only in recent years due to the accumulation of plastics, including PET, in the environment ^[6]. In 2016, Yoshida et al. ^[2] discovered the bacterium Ideonella sakaiensis 201-F6. This bacterium uses PET as its main carbon and energy source. The enzyme PETase is essential to the bacterium's growth, as its primary function is to create molecules that can be assimilated by the micro organism. Thanks to bacteria's rapid adaptation to their environment, we found a potential solution to our plastic polution problem.

Structure

Primary Structure

Secondary Structure

Tertiary Structure

Catalytic site

Disulfide bridges

Applications

Mutation of PET hydrolase

Today 349 putative PET hydrolases are identified in marine and terrestrial datasets. These PET hydrolase frequencies ranged from 0.004 to 0.92 hits/Mb and 0.0001 to 1.513 hits/Mb for marine and terrestrial datasets, respectively ^[1]. However, a metagenomic sample from a crude oil reservoir offered the highest rate of sequence hits, with a frequency about 1.5 hits/Mb ^[1][7]

Degradation of PET

As a polymer, PET is a complex structure with crystalline regions that feature tightly packed chains in parallel, and amorphous regions where the chains are disordered. However, PET has different degrees of crystallization: it is around 35% for bottles and textiles and 6% for PET used in packaging (PET film).The most important use of PET hydrolase is the degradation of PET. Although different mutants, none of these enzymes is able to dissolve all forms of PET. PET hydrolase enzymes preferentially degrade the regions of PET that are amorphous in nature because of the flexibility and movement of the polymer chains less restricted. To remedy this, the reaction takes place above the Tg of PET, which is the glass transition temperature: amorphous PET having a Tg of 67°C and crystalline PET having a Tg of 81°C ^[7].

Bioremediation

Bioremediation refers to the use of living organisms or their enzymes to detoxify or restore contaminated sites, often by directing the natural capabilities of microbes toward environmental pollutants. For example, environmental Pseudomonas isolates have been shown to degrade polyethylene by reducing the weight of high density polyethylene (HDPE) and low density polyethylene (LDPE) by 55 and 77%, respectively, following 120 days incubation. Another solution to facilitate bioremediation of ocean plastic is to use membrane-based systems featuring immobilized microbes or enzymes ( like PET hydrolase ) for plastic degradation ^[7].

Biological recycling

In mechanical recycling, collected and sorted PET waste can be powdered before melting and reprocessing to other forms. Chemical recycling leads to degrade PET into its basic monomers which can then be repolymerized ^[7]. However, this method is unfavorable because mechanical recycling is much more cost effective. Moreover, chemical methods require the maintenance of high temperature and pressure as well as employing toxic reagents and several preceding unit operations. Therefore, biological recycling is emerging as a more sustainable solution as it can be done with low temperature conditions, without the use of hazardous chemicals, by using microbial catalysis of polymer bond cleavage reactions, which results in the recovery of monomers ^[7]. However, bio-recycling is limited by the organism used, inherent polymer properties and the choice of pre-treatment, so modifications of these factors are discussed.

Circular bioeconomy

Circular economy is creating loops which feed resources back into the economy to make the same or new products. In general, the low production cost of plastic shows that the reuse does not offer an economic advantage ^[7]. However, a combination of biodegradation and biosynthesis, bio-based PET economy could contribute to an environmental advantage. A biotechnology leading to introduce PET hydrolase in the circular economy, will create PET waste and reduce its release into the environment. Bio-PET, which refers to a PET polymer that is at least partially derived from biological sources, can be produced through the microbial synthesis of terephthalic acid TPA and ethylene glycol EG ^[1]. This method could make a significant contribution to a sustainable and circular PET economy. However, some complexities are associated with biological TPA production and therefore, it is only EG that is produced biologically from renewable feedstocks to give bio-PET ^[7].

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References

1. ↑ ^{1.0 1.1 1.2 1.3} « Découverte de la PETase, une enzyme bactérienne capable de dégrader le plastique PET (polyéthylène téréphtalate) », Quoi dans mon assiette, avr. 19, 2018. https://quoidansmonassiette.fr/decouverte-petase-enzyme-bacterienne-degrader-plastique-pet-polyethylene-terephtalate/ Retrieved 2021-01-11.
2. ↑ ^{2.0 2.1} Yoshida S, Hiraga K, Takehana T, Taniguchi I, Yamaji H, Maeda Y, Toyohara K, Miyamoto K, Kimura Y, Oda K. A bacterium that degrades and assimilates poly(ethylene terephthalate). Science. 2016 Mar 11;351(6278):1196-9. doi: 10.1126/science.aad6359. PMID:26965627 doi:http://dx.doi.org/10.1126/science.aad6359
3. ↑ Panda T, Gowrishankar BS. Production and applications of esterases. Appl Microbiol Biotechnol. 2005 Apr;67(2):160-9. doi: 10.1007/s00253-004-1840-y. , Epub 2005 Jan 4. PMID:15630579 doi:http://dx.doi.org/10.1007/s00253-004-1840-y
4. ↑ P. Dockrill, « Scientists Have Accidentally Created a Mutant Enzyme That Eats Plastic Waste », ScienceAlert. https://www.sciencealert.com/scientists-accidentally-engineered-mutant-enzyme-eats-through-plastic-pet-petase-pollution Retrieved 2021-01-11.
5. ↑ Kim JW, Park SB, Tran QG, Cho DH, Choi DY, Lee YJ, Kim HS. Functional expression of polyethylene terephthalate-degrading enzyme (PETase) in green microalgae. Microb Cell Fact. 2020 Apr 28;19(1):97. doi: 10.1186/s12934-020-01355-8. PMID:32345276 doi:http://dx.doi.org/10.1186/s12934-020-01355-8
6. ↑ Austin HP, Allen MD, Donohoe BS, Rorrer NA, Kearns FL, Silveira RL, Pollard BC, Dominick G, Duman R, El Omari K, Mykhaylyk V, Wagner A, Michener WE, Amore A, Skaf MS, Crowley MF, Thorne AW, Johnson CW, Woodcock HL, McGeehan JE, Beckham GT. Characterization and engineering of a plastic-degrading aromatic polyesterase. Proc Natl Acad Sci U S A. 2018 Apr 17. pii: 1718804115. doi:, 10.1073/pnas.1718804115. PMID:29666242 doi:http://dx.doi.org/10.1073/pnas.1718804115
7. ↑ ^{7.0 7.1 7.2 7.3 7.4 7.5 7.6} Carr CM, Clarke DJ, Dobson ADW. Microbial Polyethylene Terephthalate Hydrolases: Current and Future Perspectives. Front Microbiol. 2020 Nov 11;11:571265. doi: 10.3389/fmicb.2020.571265., eCollection 2020. PMID:33262744 doi:http://dx.doi.org/10.3389/fmicb.2020.571265