PHL7 PES-H1 PESH-2 are very similar proteins PHL7 and PES-H1 are identical and PES-H2 differs in only four residues (A/E1, L/F209, D/N232, and S/A254), only the L/F209 is close to the catalytic triad. These proteins are not separated here and considered as mutants of the same entry.
(Below N is a link to NCBI taxonomic web page and E link to ESTHER at designed phylum.) > cellular organisms: NE > Bacteria: NE > Terrabacteria group: NE > Firmicutes: NE > Clostridia: NE > Thermoanaerobacterales: NE > unclassified Thermoanaerobacterales: NE > Thermoanaerobacterales bacterium: NE
Molecular evidence
Database
No mutation 11 structures(e.g. : 7CUV, 7E30, 7E31... more)(less) 7CUV: Crystal structure of a novel alpha/beta hydrolase in apo form PHL7 PES-H1, 7E30: Crystal structure of a novel alpha/beta hydrolase in apo form PHL7 PES-H1 in complex with citrate, 7E31: Crystal structure of a novel alpha/beta hydrolase mutant form PHL7 PES-H2 in complex with PEG6000, 7NEI: Novel Polyester Hydrolase Leipzig 7 (PHL-7) PES-H1, 7W66: Crystal structure of a novel alpha/beta hydrolase mutant form PHL7 PES-H2 in complex with PEG6000, 7W69: Crystal structure of a PSH1 mutant in complex with EDO PHL7 PES-H2, 7W6C: Crystal structure of a novel alpha/beta hydrolase PHL7 PES-H1 in complex with MHETA J1K, 7W6O: Crystal structure of a novel alpha/beta hydrolase PHL7 PES-H1 in complex with MHETA J1K, 7W6Q: Crystal structure of a novel alpha/beta hydrolase PHL7 PES-H1 in complex with MHETA J1K, 8BRA: Polyester Hydrolase Leipzig 7 (PHL7) bound to terephthalic acid (TPA) and Mg2+, 8BRB: Polyester Hydrolase Leipzig 7 (PHL7) bound to terephthalic acid (TPA) No kinetic
LegendThis sequence has been compared to family alignement (MSA) red => minority aminoacid blue => majority aminoacid color intensity => conservation rate title => sequence position(MSA position)aminoacid rate Catalytic site Catalytic site in the MSA MRHMSTSIARVRTRLGALLAGVIVAGSTVIGAAPAGAQENPYERGPDPTE SSIEAVRGPFAVAQTTVSRLQADGFGGGTIYYPTDTSQGTFGAVAISPGF TAGQESIAWLGPRIASQGFVVITIDTITRLDQPDSRGRQLQAALDHLRTN SVVRNRIDPNRMAVMGHSMGGGGALSAAANNTSLEAAIPLQGWHTRKNWS SVRTPTLVVGAQLDTIAPVSSHSEAFYNSLPSDLDKAYMELRGASHLVSN TPDTTTAKYSIAWLKRFVDDDLRYEQFLCPAPDDFAISEYRSTCPHSGSG GGTTPPPEDDCPWWAWWC
The recently discovered metagenomic-derived polyester hydrolase PHL7 is able to efficiently degrade amorphous polyethylene terephthalate (PET) in post-consumer plastic waste. We present the cocrystal structure of this hydrolase with its hydrolysis product terephthalic acid and elucidate the influence of 17 single mutations on the PET-hydrolytic activity and thermal stability of PHL7. The substrate-binding mode of terephthalic acid is similar to that of the thermophilic polyester hydrolase LCC and deviates from the mesophilic IsPETase. The subsite I modifications L93F and Q95Y, derived from LCC, increased the thermal stability, while exchange of H185S, derived from IsPETase, reduced the stability of PHL7. The subsite II residue H130 is suggested to represent an adaptation for high thermal stability, whereas L210 emerged as the main contributor to the observed high PET-hydrolytic activity. Variant L210T showed significantly higher activity, achieving a degradation rate of 20 microm h(-1) with amorphous PET films.
The Earth has entered the Anthropocene, which is branded by ubiquitous and devastating environmental pollution from plastics such as polyethylene terephthalate (PET). Ecofriendly and at the same time economical solutions for plastic recycling and reuse are being sought more urgently now than ever. With the possibility to recover its building blocks, the hydrolysis of PET waste by its selective biodegradation with polyester hydrolases is an appealing solution. We demonstrate how changing the dielectric properties of PET films can be used to evaluate the performance of polyester hydrolases. For this purpose, a PET film separates two reaction chambers in an impedimetric setup to quantify the film thickness- and surface area-dependent change in capacitance caused by the enzyme. The derived degradation rates determined for the polyester hydrolases PHL7 and LCC were similar to those obtained by gravimetric and vertical scanning interferometry measurements. Compared to optical methods, this technique is also insensitive to changes in the solution composition. AFM and FEM simulations further supported that impedance spectroscopy is a powerful tool for the detailed analysis of the enzymatic hydrolysis process of PET films. The developed monitoring system enabled both high-temporal resolution and parallel processing suitable for the analysis of the enzymatic degradability of polyester films and the properties of the biocatalysts.
Our planet is flooded with plastics and the need for sustainable recycling strategies of polymers has become increasingly urgent. Enzyme-based hydrolysis of post-consumer plastic is an emerging strategy for closed-loop recycling of polyethylene terephthalate (PET). The polyester hydrolase PHL7 isolated from a compost metagenome completely hydrolyzed amorphous PET films, releasing 91 mg of terephthalic acid per hour and mg of enzyme. Degradation rates of the PET film of 6.8 microm h -1 were monitored by vertical scanning interferometry. Structural analysis indicated the importance of leucine at position 210 for the extraordinarily high PET-hydrolyzing activity of PHL7. Within 24 h, 0.6 mg enzyme g PET -1 completely degraded post-consumer thermoform PET packaging in an aqueous buffer at 70 degreesC without any energy-intensive pretreatments. Terephthalic acid recovered from the enzymatic hydrolysate was used to synthesize virgin PET, demonstrating the potential of polyester hydrolases as catalysts in sustainable PET recycling processes with a low carbon footprint.
The recently discovered metagenomic-derived polyester hydrolase PHL7 is able to efficiently degrade amorphous polyethylene terephthalate (PET) in post-consumer plastic waste. We present the cocrystal structure of this hydrolase with its hydrolysis product terephthalic acid and elucidate the influence of 17 single mutations on the PET-hydrolytic activity and thermal stability of PHL7. The substrate-binding mode of terephthalic acid is similar to that of the thermophilic polyester hydrolase LCC and deviates from the mesophilic IsPETase. The subsite I modifications L93F and Q95Y, derived from LCC, increased the thermal stability, while exchange of H185S, derived from IsPETase, reduced the stability of PHL7. The subsite II residue H130 is suggested to represent an adaptation for high thermal stability, whereas L210 emerged as the main contributor to the observed high PET-hydrolytic activity. Variant L210T showed significantly higher activity, achieving a degradation rate of 20 microm h(-1) with amorphous PET films.
The Earth has entered the Anthropocene, which is branded by ubiquitous and devastating environmental pollution from plastics such as polyethylene terephthalate (PET). Ecofriendly and at the same time economical solutions for plastic recycling and reuse are being sought more urgently now than ever. With the possibility to recover its building blocks, the hydrolysis of PET waste by its selective biodegradation with polyester hydrolases is an appealing solution. We demonstrate how changing the dielectric properties of PET films can be used to evaluate the performance of polyester hydrolases. For this purpose, a PET film separates two reaction chambers in an impedimetric setup to quantify the film thickness- and surface area-dependent change in capacitance caused by the enzyme. The derived degradation rates determined for the polyester hydrolases PHL7 and LCC were similar to those obtained by gravimetric and vertical scanning interferometry measurements. Compared to optical methods, this technique is also insensitive to changes in the solution composition. AFM and FEM simulations further supported that impedance spectroscopy is a powerful tool for the detailed analysis of the enzymatic hydrolysis process of PET films. The developed monitoring system enabled both high-temporal resolution and parallel processing suitable for the analysis of the enzymatic degradability of polyester films and the properties of the biocatalysts.
Thermophilic polyester hydrolases (PES-H) have recently enabled biocatalytic recycling of the mass-produced synthetic polyester polyethylene terephthalate (PET), which has found widespread use in the packaging and textile industries. The growing demand for efficient PET hydrolases prompted us to solve high-resolution crystal structures of two metagenome-derived enzymes (PES-H1 and PES-H2) and notably also in complex with various PET substrate analogues. Structural analyses and computational modeling using molecular dynamics simulations provided an understanding of how product inhibition and multiple substrate binding modes influence key mechanistic steps of enzymatic PET hydrolysis. Key residues involved in substratebinding and those identified previously as mutational hotspots in homologous enzymes were subjected to mutagenesis. At 72 C, the L92F/Q94Y variant of PES-H1 exhibited 2.3-fold and 3.4-fold improved hydrolytic activity against amorphous PET films and pretreated real-world PET waste, respectively. The R204C/S250C variant of PES-H1 had a 6.4 C higher melting temperature than the wild-type enzyme but retained similar hydrolytic activity. Under optimal reaction conditions, the L92F/Q94Y variant of PES-H1 hydrolyzed low-crystallinity PET materials 2.2-fold more efficiently than LCC ICCG, which was previously the most active PET hydrolase reported in the literature. This property makes the L92F/ Q94Y variant of PES-H1 a good candidate for future applications in industrial plastic r"cycling processes.
Our planet is flooded with plastics and the need for sustainable recycling strategies of polymers has become increasingly urgent. Enzyme-based hydrolysis of post-consumer plastic is an emerging strategy for closed-loop recycling of polyethylene terephthalate (PET). The polyester hydrolase PHL7 isolated from a compost metagenome completely hydrolyzed amorphous PET films, releasing 91 mg of terephthalic acid per hour and mg of enzyme. Degradation rates of the PET film of 6.8 microm h -1 were monitored by vertical scanning interferometry. Structural analysis indicated the importance of leucine at position 210 for the extraordinarily high PET-hydrolyzing activity of PHL7. Within 24 h, 0.6 mg enzyme g PET -1 completely degraded post-consumer thermoform PET packaging in an aqueous buffer at 70 degreesC without any energy-intensive pretreatments. Terephthalic acid recovered from the enzymatic hydrolysate was used to synthesize virgin PET, demonstrating the potential of polyester hydrolases as catalysts in sustainable PET recycling processes with a low carbon footprint.
Thermoanaerobacterales bacterium Polyester Hydrolase Leipzig 7 (PHL-7) PHL7 PES-H1
