Enzymatic Degradation of Poly(ethylene terephthalate): Rapid Hydrolyse using a Hydrolase from <i>T. fusca</i>

Müller, Rolf‐Joachim; Schrader, Hedwig; Profe, Jörn; Dresler, Karolin; Deckwer, Wolf‐Dieter

doi:10.1002/marc.200500410

Cited by 480 publications

(366 citation statements)

References 13 publications

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“…Cutinase is one of these enzymes, and cutinases from F. solani f. sp. pisi (6,10,26,36), F. oxysporum (25), and T. fusca (1,13,14,24,32) have been well studied regarding PET modification. However, the catalytic efficiencies of these enzymes are not sufficiently high to meet the requirements of the textile industry (10).…”

Section: Resultsmentioning

confidence: 99%

Isolation of a Novel Cutinase Homolog with Polyethylene Terephthalate-Degrading Activity from Leaf-Branch Compost by Using a Metagenomic Approach

Sulaiman

Yamato

Kanaya

et al. 2012

Appl Environ Microbiol

431

385

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The gene encoding a cutinase homolog, LC-cutinase, was cloned from a fosmid library of a leaf-branch compost metagenome by functional screening using tributyrin agar plates. LC-cutinase shows the highest amino acid sequence identity of 59.7% to Thermomonospora curvata lipase. It also shows the 57.4% identity to Thermobifida fusca cutinase. When LCcutinase without a putative signal peptide was secreted to the periplasm of Escherichia coli cells with the assistance of the pelB leader sequence, more than 50% of the recombinant protein, termed LC-cutinase*, was excreted into the extracellular medium. It was purified and characterized. LC-cutinase* hydrolyzed various fatty acid monoesters with acyl chain lengths of 2 to 18, with a preference for short-chain substrates (C 4 substrate at most) most optimally at pH 8.5 and 50°C, but could not hydrolyze olive oil. It lost activity with half-lives of 40 min at 70°C and 7 min at 80°C. LC-cutinase* had an ability to degrade poly(-caprolactone) and polyethylene terephthalate (PET). The specific PET-degrading activity of LC-cutinase* was determined to be 12 mg/h/mg of enzyme (2.7 mg/h/kat of pNP-butyrate-degrading activity) at pH 8.0 and 50°C. This activity is higher than those of the bacterial and fungal cutinases reported thus far, suggesting that LC-cutinase* not only serves as a good model for understanding the molecular mechanism of PET-degrading enzyme but also is potentially applicable for surface modification and degradation of PET. C utinase (EC 3.1.1.74) is a lipolytic/esterolytic enzyme that hydrolyzes not only cutin, which is a major component of plant cuticle (38), but also water-soluble esters and insoluble triglycerides (12). It hydrolyzes these substrates to carboxylic acids and alcohols through the formation of an acyl enzyme intermediate, in which the active-site serine residue is acylated by the substrate. This serine residue is located within a GXSXG sequence motif and forms a catalytic triad with His and Asp. Cutinase has been found in both fungi and bacteria. The crystal structures of two fungal cutinases from Fusarium solani f. sp. pisi (22) and Glomerella cingulata (27) have been determined. According to these structures, cutinase shares a common ␣/␤ hydrolase fold with lipase and esterase (28). However, cutinase, like esterase, does not have a lid structure, which is responsible for interfacial activation of lipase (8). Therefore, cutinase does not show interfacial activation like esterase (14). Cutinase has recently received much attention because of its potential application for surface modification and degradation of aliphatic and aromatic polyesters (16), especially polyethylene terephthalate (PET), which is a synthetic aromatic polyester composed of terephthalic acid (TPA) and ethylene glycol (10,16,36,39). However, the number of cutinases, which have been studied regarding PET modification, is still limited, and this limitation may result in the delay of the research toward the practical use of cutinases. Therefore, isolation of a novel cutinase...

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Section: Resultsmentioning

confidence: 99%

Isolation of a Novel Cutinase Homolog with Polyethylene Terephthalate-Degrading Activity from Leaf-Branch Compost by Using a Metagenomic Approach

Sulaiman

Yamato

Kanaya

et al. 2012

Appl Environ Microbiol

431

385

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“…The authors claim that there are very few reports on the biological degradation of PET, except that a few fungal species or enzymes have been shown to be able to degrade PET (1). However, the authors did not cite three previous studies demonstrating that some bacteria or bacterial enzymes, such as cutinases, are also able to effectively degrade PET (2)(3)(4).…”

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confidence: 90%

Comment on “A bacterium that degrades and assimilates poly(ethylene terephthalate)”

Yang

Jiang

2016

Science

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Yoshida et al . (Report, 11 March 2016, p. 1196) reported that the bacterium Ideonella sakaiensis 201-F6 can degrade and assimilate poly(ethylene terephthalate) (PET). However, the authors exaggerated degradation efficiency using a low-crystallinity PET and presented no straightforward experiments to verify depolymerization and assimilation of PET. Thus, the authors’ conclusions are rather misleading.

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“…6 The enzymatic hydrolysis of PET is an environmentally friendly alternative to conventional recycling methods and can be performed under milder temperature and pH conditions, allowing less energy consumption. 7 Hydrolytic activity against PET has been identified in filamentous fungi, such as Fusarium oxysporum and Fusarium solani, 8 and in bacteria from the genus Thermobifida, 9 with enzymes such as cutinases (EC 3.1.1.74), lipases (EC 3.1.1.3), and carboxylesterases (EC 3.1.1.1) involved in PET degradation. 4 This is usually monitored by high-performance liquid chromatography (HPLC), which separates monomeric TPA from its ethylene glycol esters, namely mono(2-hydroxyethyl) terephthalate (MHET) and bis-(2-hydroxyethyl) terephthalate (BHET), among other hydrolysis products.…”

Section: Introductionmentioning

confidence: 99%

“…Additionally, a number of approaches to evaluate the extent of biodegradation have been used, including turbidimetric analysis, 10 titration, 11 image scanning, 12 and polymer film weight loss monitoring. 7 Fluorometric TPA detection has also been used to monitor PET degradation, but only for isolated enzymes. 8,13 The high sensitivity of these fluorometric methods allows the assay to be performed with low concentrations of both substrate and biocatalyst (isolated enzymes or whole-cell microorganisms), ensuring a low-cost for the screening without loss of performance.…”

Section: Introductionmentioning

confidence: 99%

A Practical Fluorescence-Based Screening Protocol for Polyethylene Terephthalate Degrading Microorganisms

Chaves¹,

Lima²,

Malafatti-Picca³

et al. 2017

J. Braz. Chem. Soc.

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We propose a practical, low-cost and selective fluorescence-based protocol adapted to identify polyethylene terephthalate (PET) degrading microorganisms. The microbial hydrolysis of PET nanoparticles was monitored by 2-hydroxyterephthalate, a fluorophore produced in situ after radical hydroxylation of terephthalic acid (TPA), the final hydrolysis product, by the Fenton reaction. Seven fungi presenting promising PET hydrolytic potential using the proposed microscale screening assay were identified. The strains evaluated presented a substantial increase of up to 18-fold in PET nanoparticles conversion, such as obtained by the fungus Trichoderma sp. C70, after their cultivation in a PET-enriched medium. The formation of other hydrolysis products, along with TPA, was observed using matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS).

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Enzymatic Degradation of Poly(ethylene terephthalate): Rapid Hydrolyse using a Hydrolase from T. fusca

Cited by 480 publications

References 13 publications

Isolation of a Novel Cutinase Homolog with Polyethylene Terephthalate-Degrading Activity from Leaf-Branch Compost by Using a Metagenomic Approach

Isolation of a Novel Cutinase Homolog with Polyethylene Terephthalate-Degrading Activity from Leaf-Branch Compost by Using a Metagenomic Approach

Comment on “A bacterium that degrades and assimilates poly(ethylene terephthalate)”

A Practical Fluorescence-Based Screening Protocol for Polyethylene Terephthalate Degrading Microorganisms

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