EP4739773A1 - Thermostable engineered enzyme - Google Patents

Thermostable engineered enzyme

Info

Publication number
EP4739773A1
EP4739773A1 EP24746059.5A EP24746059A EP4739773A1 EP 4739773 A1 EP4739773 A1 EP 4739773A1 EP 24746059 A EP24746059 A EP 24746059A EP 4739773 A1 EP4739773 A1 EP 4739773A1
Authority
EP
European Patent Office
Prior art keywords
enzyme
seq
petase
pet
engineered
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP24746059.5A
Other languages
German (de)
French (fr)
Inventor
Alfonso GAUTIERI
Emilio PARISINI
Hajar ESTIRI
Rossella CASTAGNA
Shapla BHATTACHARYA
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Politecnico di Milano
Latvian Institute of Organic Synthesis
Original Assignee
Politecnico di Milano
Latvian Institute of Organic Synthesis
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Politecnico di Milano, Latvian Institute of Organic Synthesis filed Critical Politecnico di Milano
Publication of EP4739773A1 publication Critical patent/EP4739773A1/en
Pending legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/18—Carboxylic ester hydrolases (3.1.1)
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
    • B09B—DISPOSAL OF SOLID WASTE NOT OTHERWISE PROVIDED FOR
    • B09B3/00—Destroying solid waste or transforming solid waste into something useful or harmless
    • B09B3/60—Biochemical treatment, e.g. by using enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/52—Genes encoding for enzymes or proenzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/01—Carboxylic ester hydrolases (3.1.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/01—Carboxylic ester hydrolases (3.1.1)
    • C12Y301/01074—Cutinase (3.1.1.74)
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
    • B09B—DISPOSAL OF SOLID WASTE NOT OTHERWISE PROVIDED FOR
    • B09B2101/00—Type of solid waste
    • B09B2101/75—Plastic waste
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J11/00—Recovery or working-up of waste materials
    • C08J11/04—Recovery or working-up of waste materials of polymers
    • C08J11/10—Recovery or working-up of waste materials of polymers by chemically breaking down the molecular chains of polymers or breaking of crosslinks, e.g. devulcanisation
    • C08J11/105—Recovery or working-up of waste materials of polymers by chemically breaking down the molecular chains of polymers or breaking of crosslinks, e.g. devulcanisation by treatment with enzymes
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2367/00—Characterised by the use of polyesters obtained by reactions forming a carboxylic ester link in the main chain; Derivatives of such polymers
    • C08J2367/02—Polyesters derived from dicarboxylic acids and dihydroxy compounds
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
    • Y02W30/00—Technologies for solid waste management
    • Y02W30/50—Reuse, recycling or recovery technologies
    • Y02W30/62—Plastics recycling; Rubber recycling

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Environmental & Geological Engineering (AREA)
  • Medicinal Chemistry (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Separation, Recovery Or Treatment Of Waste Materials Containing Plastics (AREA)

Abstract

The invention relates to an engineered PETase enzyme comprises an amino acid sequence having at least 70% or 80%, 90%, 95%, 99% or 100% sequence identity to SEQ ID NO: 1. Preferably, the PETase enzyme comprises SEQ ID NO.1 with at least four mutations. The invention also refers to a composition and to a method for decomposing plastics.

Description

Thermostable engineered enzyme
FIELD OF THE INVENTION
The present invention relates to a novel plastic degrading enzyme and a plastic decomposition method using the same.
STATE OF THE ART
PET (Polyethylene terephthalate) is a type of thermoplastic made by polymerization of TPA (terephthalic acid) and EG (ethylene glycol). PET has high transparency and excellent thermal insulation properties, so it is a polymer material commonly used in electric wire coverings, household goods, toys, and packaging materials, and is particularly widely used in the manufacture of bottles. However, since PET has a chemical structure that is not easily decomposed in nature, environmental pollution caused by the accumulation of PET in ecosystems including soil, rivers, and oceans is recognized as a serious social problem. In particular, the bioaccumulation problem caused by microplastics can adversely affect human health, so the need for recycling or environmentally friendly decomposition of PET is gradually increasing.
However, when PET is recycled industrially, the recycling rate is not high due to poor quality and high cost, and when PET is decomposed through physical and chemical methods, endocrine-disrupting chemicals such as dioxins can be produced, which is a secondary problem. There is a problem that occurs.
In 1977, Tokiwa and Suzuki proposed the idea of using enzymes to degrade polymeric materials. Indeed, enzymes work in mild conditions and can replace hazardous chemicals (a concept known as green chemistry). Since then, many PET-degrading enzymes from various microorganisms have been discovered and characterized, which has also led to the implementation of a series of strategies for enhancing their catalytic properties. Despite the availability of many suitable enzymes, their practical use for PET degradation is limited, mostly due to their low thermal stability. Indeed, the enzymatic degradation of PET by PETases does improve at high temperatures, where PET crystallinity is reduced. Hence, the design of a PETase with enhanced thermal stability is particularly relevant to the development of enzymatic PET-degrading strategies.
SUMMARY OF THE INVENTION
The first aspect of the invention relates to an engineered PETase enzyme. In one embodiment, the engineered PETase enzyme comprises an amino acid sequence having at least 70% or 80%, 90%, 95%, 99% or 100% sequence identity to SEQ ID NO: 1 .
Preferably, the PETase enzyme comprising SEQ ID NO.1 with at least four mutations, also referred as amino acid substitutions. In one embodiment, said SEQ ID NO.1 comprises at least four mutations selected from: S241 D, S36D, Q40R, S48D, S57K, S145R, A209R, N276D, N278K and Q293K.
A second aspect of the present invention refers to a composition for decomposing plastics.
The composition for decomposing plastic comprises the engineered PETase enzyme comprises an amino acid sequence having at least 70% or 80%, 90%, 95%, 99% or 100% sequence identity to SEQ ID NO: 2 and/or the engineered PETase enzyme comprises an amino acid sequence having at least 70% or 80%, 90%, 95%, 99% or 100% sequence identity to SEQ ID NO: 3.
Another aspect of the present invention provides a method for decomposing plastics.
The method for decomposing plastics includes treating the plastics with the composition for decomposing plastics of the present invention. The plastic may be MHET, BHET, PET, or the like, or derivatives thereof.
BRIEF DESCRIPTION OF THE FIGURES
The Figures show a comparison of PET-depolymerization over time for C09, C08 and ICCG (40 nM, 80 °C, pH 8). Means ± s.d (n=2) are shown. Figure 1 shows TPA production over time;
Figure 2 shows TPA production at 4h and 144h. *P<0.05; **P<0.01 (one- side unpaired Welch’s t-test); and
Figure 3 shows enzyme specific activity at the different time points.
DETAILED DESCRIPTION OF THE INVENTION
The first aspect of the invention relates to an engineered PETase enzyme. The term “PETase” or “polyethylene terephthalate hydrolases” refer to a class of hydrolases that have the property of catalysing the cleavage of PET (polyethylene terephthalate) into mono (2-hydroxyethyl) terephthalic acid.
In one embodiment, there is provided an engineered thermostable PETase enzyme with a melting temperature (Tm) comprised between 80°C and 100°C, preferably, comprised between 82 and 98°C, that is higher than wild type enzyme. The engineered PETase enzyme may be active at higher temperatures (80-90 °C) enabling more efficient degradation of substrates. The thermostable PETase enzyme may also have longer shelf life at ambient temperatures.
In one embodiment, the PETase enzyme has improved thermal stability. In one embodiment, the engineered PETase enzyme comprises an amino acid sequence having at least 70% or 80%, 90%, 95%, 99% or 100% sequence identity to SEQ ID NO: 1 .
Preferably, the PETase enzyme comprises SEQ ID NO.1 with at least four mutations, also referred as amino acid substitutions. In one embodiment, said SEQ ID NO.1 comprises at least four mutations selected from: S241 D, S36D, Q40R, S48D, S57K, S145R, A209R, N276D, N278K and Q293K. In other words, the enzyme comprises SEQ ID NO. 1 with at least four mutations in the above-identified positions.
In some embodiments, SEQ ID NO.1 comprises at least five mutations selected from: S241 D, S36D, Q40R, S48D, S57K, S145R, A209R, N276D, N278K and Q293K. Preferably, SEQ ID NO.1 comprises at least six mutations selected from: S241 D, S36D, Q40R, S48D, S57K, S145R, A209R, N276D, N278K and Q293K.
In some embodiments, SEQ ID NO.1 comprises at least seven mutations selected from: S241 D, S36D, Q40R, S48D, S57K, S145R, A209R, N276D, N278K and Q293K. Preferably, SEQ ID NO. 1 comprises the mutations: S48D, S57K, S145R, A209R, S241 D, N276D, N278K, Q293K.
In one embodiment, the engineered PETase enzyme comprises an amino acid sequence having at least 70% or 80%, 90%, 95%, 99% or 100% sequence identity to SEQ ID NO: 2.
In a preferred embodiment, the PETase enzyme consists of SEQ ID NO.2. Preferably, SEQ ID NO. 1 comprises the mutations: S36D, Q40R, S48D, S57K, S145R, A209R, N276D, N278K, Q293K.
In one embodiment, the engineered PETase enzyme comprises an amino acid sequence having at least 70% or 80%, 90%, 95%, 99% or 100% sequence identity to SEQ ID NO: 3.
In a preferred embodiment, the PETase enzyme consists of SEQ ID NO.3.
Another aspect of the present invention provides a composition for decomposing plastics. The composition for decomposing plastic comprises at least one PTease enzyme as detailed disclosed above. Preferably, the composition comprises the engineered PETase enzyme comprises an amino acid sequence having at least 70% or 80%, 90%, 95%, 99% or 100% sequence identity to SEQ ID NO: 2 and/or the engineered PETase enzyme comprises an amino acid sequence having at least 70% or 80%, 90%, 95%, 99% or 100% sequence identity to SEQ ID NO: 3.
In a preferred embodiment, the composition comprises the engineered PETase enzyme consisting of SEQ ID NO: 2 and/or the engineered PETase enzyme consisting of SEQ ID NO: 3. The plastic may be Mono-(2-hydroxyethyl)terephthalic acid (MHET) and Bis-(2-hydroxyethyl)terephthalic acid (BHET), and PET, or the like, or a derivative thereof. In some embodiments, the composition for decomposing plastics of the present invention comprises at least one ion selected from the group consisting of Ni2+, Fe2+, Co2+, Cu2+, Mg2+, Mn2+, K+, Zn2+ and EDTA, preferably at a concentration of 10 mM or less in the composition of the present invention.
Another aspect of the present invention provides a method for decomposing plastics.
The method for decomposing plastics includes treating the plastics with the enzyme and/or with the composition for decomposing plastics of the present invention.
Preferably, the plastic is selected from: MHET, BHET, PET, or the like, or derivatives thereof.
The step of treating the composition on the plastic may be carried out under the conditions of pH 7.5 to pH 10.5. Specifically, the pH may be in the range of pH 7.0 to pH 10.5, 7.5 to pH 10.0, or 7.5 to pH 9.0. The plastic degrading enzyme of the present invention can exhibit a higher activity in the above pH range, thereby exhibiting an activity of 80% or more of the maximum activity.
Preferably, the enzyme is used in a concentration comprised between 20 and 60 nM, more preferably comprised between 30 and 50 nM.
Example
Enzyme stability
To this end, the Applicant focused on the so-called leaf-branch compost cutinase (LCC), a naturally occurring PETase that has been reported to outperform all other known PET-degrading enzymes and presents a melting temperature (Tm) of 84.7°C. This enzyme has been previously engineered by Tournier et al. (Nature, 2020), leading to the ICCG variant with a reported Tm of 94.5°C (88.9°C in our own assessment). Starting from this ICCG variant, different enzyme design strategies have been applied to engineer new enzyme mutants. This led to two variants of the enzyme (named C08 and C09), one of which show a Tm higher than any other PETase currently known.
Enzyme Melting temperature [Tm, °C] Improvement
LCC (Wild type) 84.7
ICCG (Tournier et al.) 94.5 (reported)* +9.8*
88.9 (our assessment)* +4.2*
C08 (this work) 82.2
C09 (this work) 96.8 +12.1
*The estimation of Tm is sensitive to the experimental setup and conditions, which could explain the difference between the reported Tm and the Tm determined by us. For proper comparison, we assessed the Tm of ICCG using the same setup that we used for the measurement of the Tm of our engineered enzymes.
Enzyme activity
Terephthalic acid (TPA) is one of the major degradation product of the activity of the enzymes on PET. This compound can be detected by bulk UV spectrophotometry allowing a precise quantification of product formation. Sample were harvested at multiple time points and analysed by ultra-high-performance liquid chromatography UHPLC for the quantification of TPA production. An enzyme concentration of 40 nM has been used to evaluate the PET degradation activity of the different mutants. The experiments were carried at 80°C and were monitored for 6 days (144h). For the selected mutants (i.e. , C09 and C08) the mean value of TPA concentration at the different time points is significantly higher for the mutated enzymes than for ICCG (Figure 1 and 2). In particular, the specific activity of C09 is much higher than the ICCG gold standard and, although it degrades over time, the mutant retains higher activity up until day 6 (144h, Figure 3).

Claims

1. An engineered PETase enzyme comprising SEQ ID NO.1 or a composition comprising it, wherein SEQ ID NO. 1 comprises at least four mutations selected from: S241 D, S36D, Q40R, S48D, S57K, S145R, A209R, N276D, N278K and Q293K.
2. The enzyme according to claim 1 , wherein SEQ ID NO.1 comprises at least five mutations selected from: S241 D, S36D, Q40R, S48D, S57K, S145R, A209R, N276D, N278K and Q293K.
3. The enzyme according to claim 1 or 2, wherein SEQ ID NO.1 comprises at least six mutations selected from: S241 D, S36D, Q40R, S48D, S57K, S145R, A209R, N276D, N278K and Q293K.
4. The enzyme according to anyone of claims 1-3, wherein SEQ ID NO.1 comprises at least seven mutations selected from: S241 D, S36D, Q40R, S48D, S57K, S145R, A209R, N276D, N278K and Q293K.
5. The enzyme according to anyone of claims 1-4, wherein SEQ ID NO. 1 comprises the mutations: S48D, S57K, S145R, A209R, S241 D, N276D, N278K, Q293K.
6. The enzyme according to claim 5, consisting of SEQ ID NO.2.
7. The enzyme according to anyone of claims 1-4, wherein SEQ ID NO. 1 comprises the mutations: S36D, Q40R, S48D, S57K, S145R, A209R, N276D, N278K, Q293K.
8. The enzyme according to claim 7 consisting of SEQ ID NO.3.
9. A method for decomposing plastics comprises at least one step of treating the plastics with the enzyme and/or with the composition according to anyone of claims 1-8.
10. The method according to claim 9, wherein the plastic is selected from: MHET, BHET, PET, preferably is PET.
11 . The method according to claim 9, or 10 wherein the enzyme is used in a concentration comprised between 20 and 60 nM, preferably comprised between 30 and 50 nM.
EP24746059.5A 2023-07-07 2024-07-08 Thermostable engineered enzyme Pending EP4739773A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
IT102023000014223A IT202300014223A1 (en) 2023-07-07 2023-07-07 THERMOSTABLE ENGINEERED ENZYME
PCT/IB2024/056641 WO2025012799A1 (en) 2023-07-07 2024-07-08 Thermostable engineered enzyme

Publications (1)

Publication Number Publication Date
EP4739773A1 true EP4739773A1 (en) 2026-05-13

Family

ID=88207206

Family Applications (1)

Application Number Title Priority Date Filing Date
EP24746059.5A Pending EP4739773A1 (en) 2023-07-07 2024-07-08 Thermostable engineered enzyme

Country Status (3)

Country Link
EP (1) EP4739773A1 (en)
IT (1) IT202300014223A1 (en)
WO (1) WO2025012799A1 (en)

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3485007B1 (en) * 2016-07-12 2025-10-29 Carbios Novel esterases and uses thereof
PT3830254T (en) * 2018-07-27 2025-01-24 Carbios Novel esterases and uses thereof
CA3107560A1 (en) * 2018-07-27 2020-01-30 Carbios Esterases and uses thereof
CN115125225B (en) * 2021-03-25 2023-07-04 湖北大学 PET degrading enzymes with improved thermostability
MX2024001814A (en) * 2021-08-11 2024-03-01 Biometis Tech Inc Enzymatic degradation of polyethylene terephthalate.
EP4433582A2 (en) * 2021-11-16 2024-09-25 Carbios Esterases and uses thereof
CN116286727B (en) * 2023-03-06 2025-09-16 山东大学 Cutinase variant, preparation method thereof and application thereof in plastic degradation

Also Published As

Publication number Publication date
WO2025012799A1 (en) 2025-01-16
IT202300014223A1 (en) 2025-01-07

Similar Documents

Publication Publication Date Title
Kawai et al. Current knowledge on enzymatic PET degradation and its possible application to waste stream management and other fields
Quartinello et al. Synergistic chemo‐enzymatic hydrolysis of poly (ethylene terephthalate) from textile waste
Thachnatharen et al. The waste management of polyethylene terephthalate (PET) plastic waste: A review
Guo et al. Natural and engineered enzymes for polyester degradation: a review
Hu et al. Enzymatic degradation of poly (butylene succinate) by cutinase cloned from Fusarium solani
Zhang et al. Catalytic amounts of an antibacterial monomer enable the upcycling of poly (ethylene terephthalate) waste
Serrano‐Aguirre et al. Can bioplastics always offer a truly sustainable alternative to fossil‐based plastics?
Aguiar et al. Enhancing plastic waste recycling: Evaluating the impact of additives on the enzymatic polymer degradation
Jiang et al. Novel polyurethane-degrading cutinase BaCut1 from Blastobotrys sp. G-9 with potential role in plastic bio-recycling
CN111718569B (en) Method for recovering polyglycolic acid and application thereof
AU2014252165A1 (en) Compositions and methods for biodegrading plastic
CN103642015B (en) A kind of Graphene oxide/aliphatic-aromcopolyester copolyester and its preparation method
CN103668539A (en) Antistatic biomass polyester fiber and preparation method thereof
CN111039864A (en) A kind of preparation method of antioxidant
Weiland Enzymatic biodegradation by exploring the rational protein engineering of the polyethylene terephthalate hydrolyzing enzyme PETase from Ideonella sakaiensis 201-F6
WO2025012799A1 (en) Thermostable engineered enzyme
Schwab et al. Correlation of enzymatic depolymerization rates with the structure of polyethylene-like long-chain aliphatic polyesters
Gomes et al. Lessons from biomass valorization for improving plastic-recycling enzymes
CN112898546A (en) Preparation method of PBAT material with high thermal stability
CN118852726A (en) A chemical-biological cascade catalytic degradation method for polyolefin plastics
CN112608568A (en) Degradable plastic and preparation method thereof
US20220243031A1 (en) Matte polyester film and method for manufacturing the same
US20230193218A1 (en) Use of lytic polysaccharide monooxygenases, enzymatic composition containing same, and degradation method for plastic polymers
CN121086338B (en) Application of degradative enzyme TfCut-B3 in depolymerizing waste plastic film
US20240052095A1 (en) Manufacturing method of thermoplastic polyester elastomer

Legal Events

Date Code Title Description
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: UNKNOWN

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE

PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE

17P Request for examination filed

Effective date: 20260203

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC ME MK MT NL NO PL PT RO RS SE SI SK SM TR