Production of extracellular PETase from Ideonella sakaiensis using sec-dependent signal peptides in E. coli

Seo, Hogyun; Kim, Seong–Min; Son, Hyeoncheol Francis; Sagong, Hye-Young; Joo, Seongjoon; Kim, Kyungjin

doi:10.1016/j.bbrc.2018.11.087

Cited by 106 publications

(62 citation statements)

References 17 publications

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“…2c). Moreover, we found that 20 of the newly identified oPETases, many of which had the M5 motif, already displayed residue substitutions that were identified as enhancing PETase activity in engineered laboratory variants of I. sakaiensis PETases (18) (Suppl . Table S3).…”

Section: Depth (M)mentioning

confidence: 96%

Rapid Evolution of Plastic-degrading Enzymes Prevalent in the Global Ocean

Álam

Gasol

Arold

et al. 2020

Preprint

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Estimates of marine plastic stocks, a major threat to marine life (1), are far lower than expected from exponentially-increasing litter inputs, suggesting important loss factors (2, 3). These may involve microbial degradation, as the plastic-degrading polyethylene terephthalate enzyme (PETase) has been reported in marine microbial communities (4). An assessment of 416 metagenomes of planktonic communities across the global ocean identifies 68 oceanic PETase variants (oPETase) that evolved from ancestral enzymes degrading polycyclic aromatic hydrocarbons. Twenty oPETases show predicted efficiencies comparable to those of laboratory-optimized PETases, suggesting strong selective pressures directing the evolution of these enzymes. We found oPETases in 90.1% of samples across all oceans and depths, particularly abundant at 1,000 m depth, with a strong dominance of Pseudomonadales containing putative highly-efficient oPETase variants in the dark ocean. Enzymatic degradation may be removing plastic from the marine environment while providing a carbon source for bathypelagic microbial communities.

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Section: Depth (M)mentioning

confidence: 96%

Rapid Evolution of Plastic-degrading Enzymes Prevalent in the Global Ocean

Álam

Gasol

Arold

et al. 2020

Preprint

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“…Follow-up investigations of PETase have determined its structure by X-ray crystallography and identified significant amino acid residues for activity, as well as made initial attempts to enhance activity (Han et al, 2017;Joo et al, 2018;Austin et al, 2018;Fecker et al, 2018;Liu et al, 2018;Chen et al, 2018;Ma et al, 2018). PET degradation using heterologously expressed PETase has been successfully shown in a variety of microorganisms, including in E. coli (Seo et al, 2019), B. subtilis (Huang et al, 2018), yeast (P. pastoris) (Z. Chen et al, 2020), and marine microalgae (P. tricornutum) (Moog et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

“…Upon incubation with various forms of PET, purified PETase has been shown either to release detectable amounts of monomers or to modify the surface of PET samples, depending on the study. The various forms of PET in which such monomer-release or surfacemodification by PETase has been demonstrated include undefined 'pet film' (Han et al, 2017;Joo et al, 2018;Seo et al, 2019), plastic bottles (this work, Yoshida et al, 2016;Liu et al, 2018;Moog et al, 2019;Chen et al, 2020), lab-synthesized amPET and hcPET (Austin et al, 2018), amPET:film (Goodfellow) (this work, Huang et al, 2018) and hcPET:film (biaxially oriented Goodfellow) (this work, Ma et al, 2018). This makes comparisons of purified PETase efficacy challenging.…”

Section: Introductionmentioning

confidence: 99%

The highly crystalline PET found in plastic water bottles does not support the growth of the PETase‐producing bacterium Ideonella sakaiensis

Wallace

Adams

Chafin

et al. 2020

Environ Microbiol Rep

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Summary Ideonella sakaiensis produces an enzyme, PETase, that is capable of hydrolyzing polyethylene terephthalate (PET) plastic. We demonstrate that although I. sakaiensis can grow on amorphous plastic, it does not grow on highly crystalline plastic under otherwise identical conditions. Both amorphous film and amorphous plastic obtained from commercial food containers support the growth of the bacteria, whereas highly crystalline film and the highly crystalline body of a plastic water bottle do not support growth. Highly crystalline PET can be melted and rapidly cooled to make amorphous plastic which then supports bacterial growth, whereas the same plastic can be melted and slowly cooled to make crystalline plastic which does not support growth. We further subject a plastic water bottle to a top‐to‐bottom analysis, finding that only amorphous sections are degraded, namely the finish (threading), the topmost portion of the shoulder which connects to the finish, and the area immediately surrounding the centre of the base. Finally, we use these results to estimate that the percentage of non‐degradable plastic in plastic water bottles ranges from 52% to 82% (depending on size), demonstrating that most of the plastic found in PET water bottles will not be degraded by I. sakaiensis.

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“…The genetically engineered microbes could be designed for multiple enzyme production, regulating of quorum sensing mechanism for biofilm formation, etc . For evidence, genetic engineering makes it doable to express PETase, a key enzyme for PET degradation in several bacterial cell-system like Bacillus subtilis and Escherichia coli [ 99 , 100 ], other than its natural host Ideonella sakaiensis [ 33 ]. The 3.8-fold expression of ‘active’ PETase enzyme was achieved in Bacillus subtilis via the Tat-independent secretory pathway using native signal peptide [ 99 ], where in Escherichia coli, PETase enzyme was expressed extracellularly via secretary (Sec) pathway-dependent manner [ 100 ].…”

Section: Biotechnological Implication Of Metagenomic Informationmentioning

confidence: 99%

“…For evidence, genetic engineering makes it doable to express PETase, a key enzyme for PET degradation in several bacterial cell-system like Bacillus subtilis and Escherichia coli [ 99 , 100 ], other than its natural host Ideonella sakaiensis [ 33 ]. The 3.8-fold expression of ‘active’ PETase enzyme was achieved in Bacillus subtilis via the Tat-independent secretory pathway using native signal peptide [ 99 ], where in Escherichia coli, PETase enzyme was expressed extracellularly via secretary (Sec) pathway-dependent manner [ 100 ]. The functionality of these heterogeneously expressed proteins was checked through PET film degradation assay [ 99 , 100 ].…”

Section: Biotechnological Implication Of Metagenomic Informationmentioning

confidence: 99%

Metagenomic Exploration of Plastic Degrading Microbes for Biotechnological Application

Purohit

Chattopadhyay

Teli

2020

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: Since the last few decades, the promiscuous and uncontrolled use of plastics leads to the accumulation of millions of tons of plastic waste in the terrestrial and marine environment and elevated the risk of environmental pollution and climate change. The concern arises more due to the reckless and unscientific disposal of plastics containing high molecular weight polymers, viz., polystyrene, polyamide, polyvinylchloride, polypropylene, polyurethane, and polyethylene, etc. which are very difficult to degrade. Thus, the focus is now paid to search for efficient, eco-friendly, low-cost waste management technology. Of them, degradation of non-degradable synthetic polymer using diverse microbial agents, viz., bacteria, fungi, and other extremophiles become an emerging option. So far, very few microbial agents and their secreted enzymes have been identified and characterized for plastic degradation, but with low efficiency. It might be due to the predominance of uncultured microbial species; consequently remain unexplored from the respective plastic degrading milieu. To overcome this problem, metagenomic analysis of microbial population engaged in the plastic biodegradation is advisable to decipher the microbial community structure and to predict their biodegradation potential in situ. Advancements in sequencing technologies and bioinformatics analysis allow the rapid metagenome screening that helps in the identification of total microbial community and also opens up the scope for mining genes or enzymes (hydrolases, laccase, etc.) engaged in polymer degradation. Further, the extraction of the core microbial population and their adaptation, fitness, and survivability can also be deciphered through comparative metagenomic study. It will help to engineer the microbial community and their metabolic activity to speed up the degradation process.

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Production of extracellular PETase from Ideonella sakaiensis using sec-dependent signal peptides in E. coli

Cited by 106 publications

References 17 publications

Rapid Evolution of Plastic-degrading Enzymes Prevalent in the Global Ocean

Rapid Evolution of Plastic-degrading Enzymes Prevalent in the Global Ocean

The highly crystalline PET found in plastic water bottles does not support the growth of the PETase‐producing bacterium Ideonella sakaiensis

Metagenomic Exploration of Plastic Degrading Microbes for Biotechnological Application

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