Manufacturing of the highly active thermophile PETases PHL7 and PHL7mut3 using Escherichia coli.
E. coli
Bioreactor cultivation
Enzyme production
PET degradation
PETase
PHL7
Recycling
Journal
Microbial cell factories
ISSN: 1475-2859
Titre abrégé: Microb Cell Fact
Pays: England
ID NLM: 101139812
Informations de publication
Date de publication:
10 Oct 2024
10 Oct 2024
Historique:
received:
12
07
2024
accepted:
30
09
2024
medline:
11
10
2024
pubmed:
11
10
2024
entrez:
10
10
2024
Statut:
epublish
Résumé
The global plastic waste crisis requires combined recycling strategies. One approach, enzymatic degradation of PET waste into monomers, followed by re-polymerization, offers a circular economy solution. However, challenges remain in producing sufficient amounts of highly active PET-degrading enzymes without costly downstream processes. Using the growth-decoupled enGenes e In this research, we present an optimized process for the extracellular production of thermophile and highly active PETases PHL7 and PHL7mut3, eliminating the need for costly purification steps. These advancements support large-scale enzymatic recycling, contributing to solving the global plastic waste crisis.
Sections du résumé
BACKGROUND
BACKGROUND
The global plastic waste crisis requires combined recycling strategies. One approach, enzymatic degradation of PET waste into monomers, followed by re-polymerization, offers a circular economy solution. However, challenges remain in producing sufficient amounts of highly active PET-degrading enzymes without costly downstream processes.
RESULTS
RESULTS
Using the growth-decoupled enGenes e
CONCLUSIONS
CONCLUSIONS
In this research, we present an optimized process for the extracellular production of thermophile and highly active PETases PHL7 and PHL7mut3, eliminating the need for costly purification steps. These advancements support large-scale enzymatic recycling, contributing to solving the global plastic waste crisis.
Identifiants
pubmed: 39390547
doi: 10.1186/s12934-024-02551-6
pii: 10.1186/s12934-024-02551-6
doi:
Substances chimiques
Polyethylene Terephthalates
0
Bacterial Proteins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
272Subventions
Organisme : Bio-based Industries Joint Undertaking (BBI JU) and the European Union's Horizon 2020 program
ID : 887913
Informations de copyright
© 2024. The Author(s).
Références
Liu C, Shi C, Zhu S, Wei R, Yin CC. Structural and functional characterization of polyethylene terephthalate hydrolase from Ideonella sakaiensis. Biochem Biophys Res Commun. 2019;508(1):289–94. https://doi.org/10.1016/j.bbrc.2018.11.148 .
doi: 10.1016/j.bbrc.2018.11.148
pubmed: 30502092
Rex Whinfield J, Tennant Dickson J. ‘Polymeric Linear Terephthalic Esters’, Public Law, vol. 1, pp. 1–7, 1945, [Online]. Available: https://www.google.com/patents/US2465319
Wyeth NC, Pa. Mendenhall, and, Roseveare RN. ‘Biaxially oriented poly(ethylene terephthalate) bottle’, United State Patent Office, pp. 1–6, 1973.
Zhang F, et al. Current technologies for plastic waste treatment: a review. J Clean Prod. 2021;282:124523. https://doi.org/10.1016/j.jclepro.2020.124523 .
doi: 10.1016/j.jclepro.2020.124523
Thomsen TB, Almdal K, Meyer AS. ‘Significance of poly(ethylene terephthalate) (PET) substrate crystallinity on enzymatic degradation’, N Biotechnol, vol. 78, no. August 2023, pp. 162–172, 2023, https://doi.org/10.1016/j.nbt.2023.11.001
Wojnowska-Baryła K, Bernat, Zaborowska M. Plastic Waste Degradation in Landfill conditions: the Problem with Microplastics, and their direct and Indirect Environmental effects. Int J Environ Res Public Health. Oct. 2022;19(20). https://doi.org/10.3390/ijerph192013223 .
Lamtai S, Elkoun M, Robert F, Mighri, Diez C. ‘Mechanical Recycling of Thermoplastics: A Review of Key Issues’, Waste, vol. 1, no. 4, pp. 860–883, Oct. 2023, https://doi.org/10.3390/waste1040050
Babaei M, Jalilian M, Shahbaz K. Chemical recycling of polyethylene terephthalate: a mini-review. J Environ Chem Eng. p. Mar. 2024;112507. https://doi.org/10.1016/j.jece.2024.112507 .
Tournier V, et al. An engineered PET depolymerase to break down and recycle plastic bottles. Nature. 2020;580(7802):216–9. https://doi.org/10.1038/s41586-020-2149-4 .
doi: 10.1038/s41586-020-2149-4
pubmed: 32269349
Soong YHV, Sobkowicz MJ, Xie D. ‘Recent Advances in Biological Recycling of Polyethylene Terephthalate (PET) Plastic Wastes’, Mar. 01, 2022, MDPI. https://doi.org/10.3390/bioengineering9030098
Maurya A, Bhattacharya, Khare SK. ‘Enzymatic remediation of polyethylene terephthalate (PET)–Based polymers for Effective Management of Plastic Wastes: an overview’, Nov. 19, 2020. Front Media S A https://doi.org/10.3389/fbioe.2020.602325
Soong YHV et al. Dec., ‘Enzyme selection, optimization, and production toward biodegradation of post-consumer poly(ethylene terephthalate) at scale’, Biotechnol J, vol. 18, no. 12, 2023, https://doi.org/10.1002/biot.202300119
Ronkvist ÅM, Xie W, Lu W, Gross RA. ‘Cutinase-Catalyzed hydrolysis of poly(ethylene terephthalate)’, Macromolecules, vol. 42, no. 14, pp. 5128–5138, Jul. 2009, https://doi.org/10.1021/ma9005318
Wei R, Song C, Gräsing D. and ‘Conformational fitting of a flexible oligomeric substrate does not explain the enzymatic PET degradation’, Nat Commun, vol. 10, no. 5581, Dec. 2019, [Online]. Available: https://doi.org/10.1038/s41467-019-13492-9
Müller RJ, Schrader H, Profe J, Dresler K, Deckwer WD. ‘Enzymatic degradation of poly(ethylene terephthalate): Rapid hydrolyse using a hydrolase from T. fusca’, Macromol Rapid Commun, vol. 26, no. 17, pp. 1400–1405, Sep. 2005, https://doi.org/10.1002/marc.200500410
Wei R et al. Aug., ‘Engineered bacterial polyester hydrolases efficiently degrade polyethylene terephthalate due to relieved product inhibition’, Biotechnol Bioeng, vol. 113, no. 8, pp. 1658–1665, 2016, https://doi.org/10.1002/bit.25941
Sonnendecker C et al. May., ‘Cover Feature: Low Carbon Footprint Recycling of Post-Consumer PET Plastic with a Metagenomic Polyester Hydrolase (ChemSusChem 9/2022)’, ChemSusChem, vol. 15, no. 9, 2022, https://doi.org/10.1002/cssc.202200696
Richter PK, et al. Structure and function of the metagenomic plastic-degrading polyester hydrolase PHL7 bound to its product. Nat Commun. Dec. 2023;14(1). https://doi.org/10.1038/s41467-023-37415-x .
Tiong E, et al. Expression and engineering of PET-degrading enzymes from Microbispora, Nonomuraea, and Micromonospora. Appl Environ Microbiol. 2023;89(11). https://doi.org/10.1128/aem.00632-23 .
Van Gemeren A, Beijersbergen A, Van Den Hondel CAMJJ, Verrips CT. Expression and secretion of defined cutinase variants by aspergillus awamori. Appl Environ Microbiol. 1998;64(8):2794–9.
doi: 10.1128/AEM.64.8.2794-2799.1998
pubmed: 9687432
pmcid: 106774
Wang X, Song C, Qi Q, Zhang Y, Li R, Huo L. Biochemical characterization of a polyethylene terephthalate hydrolase and design of high-throughput screening for its directed evolution. Eng Microbiol. Jun. 2022;2(2). https://doi.org/10.1016/j.engmic.2022.100020 .
Sulaiman S, et al. Isolation of a novel cutinase homolog with polyethylene terephthalate-degrading activity from leaf-branch compost by using a metagenomic approach. Appl Environ Microbiol. 2012;78(5):1556–62. https://doi.org/10.1128/AEM.06725-11 .
doi: 10.1128/AEM.06725-11
pubmed: 22194294
pmcid: 3294458
Seo H, Kim S, Son HF, Sagong HY, Joo S, Kim KJ. Production of extracellular PETase from Ideonella sakaiensis using sec-dependent signal peptides in E. Coli. Biochem Biophys Res Commun. 2019;508(1):250–5. https://doi.org/10.1016/j.bbrc.2018.11.087 .
doi: 10.1016/j.bbrc.2018.11.087
pubmed: 30477746
Aer L, Jiang Q, Gul I, Qi Z, Feng J, Tang L. Overexpression and kinetic analysis of Ideonella sakaiensis PETase for polyethylene terephthalate (PET) degradation. Environ Res. Sep. 2022;212. https://doi.org/10.1016/j.envres.2022.113472 .
Su L, Xu C, Woodard RW, Chen J, Wu J. ‘A novel strategy for enhancing extracellular secretion of recombinant proteins in Escherichia coli’, Appl Microbiol Biotechnol, vol. 97, no. 15, pp. 6705–6713, Aug. 2013, https://doi.org/10.1007/s00253-013-4994-7
Su L, Jiang Q, Yu L, Wu J. Enhanced extracellular production of recombinant proteins in Escherichia coli by co-expression with Bacillus cereus phospholipase C. Microb Cell Fact. Feb. 2017;16(1). https://doi.org/10.1186/s12934-017-0639-3 .
Tan Y, Henehan GT, Kinsella GK, Ryan BJ. ‘Extracellular secretion of a cutinase with polyester-degrading potential by E. coli using a novel signal peptide from Amycolatopsis mediterranei’, World J Microbiol Biotechnol, vol. 38, no. 4, Apr. 2022, https://doi.org/10.1007/s11274-022-03246-z
Lu H, et al. Machine learning-aided engineering of hydrolases for PET depolymerization. Nature. Apr. 2022;604(7907):662–7. https://doi.org/10.1038/s41586-022-04599-z .
Bell EL et al. ‘Directed Evolution of an Efficient and Thermostable PET Depolymerase’, Nat Catal, vol. 5, pp. 673–681, 2022, [Online]. Available: https://doi.org/10.1038/s41929-022-00821-3
Son HF et al. Apr., ‘Rational Protein Engineering of Thermo-Stable PETase from Ideonella sakaiensis for Highly Efficient PET Degradation’, ACS Catal, vol. 9, no. 4, pp. 3519–3526, 2019, https://doi.org/10.1021/acscatal.9b00568
Sagong HY, et al. Structural and functional characterization of an auxiliary domain-containing PET hydrolase from Burkholderiales bacterium. J Hazard Mater. May 2022;429. https://doi.org/10.1016/j.jhazmat.2022.128267 .
Pirillo V, Orlando M, Battaglia C, Pollegioni L, Molla G. ‘Efficient polyethylene terephthalate degradation at moderate temperature: a protein engineering study of LC-cutinase highlights the key role of residue 243’, FEBS Journal, vol. 290, no. 12, pp. 3185–3202, Jun. 2023, https://doi.org/10.1111/febs.16736
Rosano GL, Ceccarelli EA. ‘Recombinant protein expression in Escherichia coli: advances and challenges’, 2014. Front Res Foundation. https://doi.org/10.3389/fmicb.2014.00172
Stargardt P, Feuchtenhofer L, Cserjan-Puschmann M, Striedner G, Mairhofer J. ‘Bacteriophage Inspired Growth-Decoupled Recombinant Protein Production in Escherichia coli’, ACS Synth Biol, vol. 9, no. 6, pp. 1336–1348, Jun. 2020, https://doi.org/10.1021/acssynbio.0c00028
Stargardt P, Striedner G, Mairhofer J. Tunable expression rate control of a growth-decoupled T7 expression system by l-arabinose only. Microb Cell Fact. Dec. 2021;20(1). https://doi.org/10.1186/s12934-021-01512-7 .
Wei S-Q, Staders J. ‘Distinct Regions of the LamB Signal Sequence Function in Different Steps in Export’, Journal of Biological Chemistry, vol. 269, no. 3, pp. 1648–1653, 1994, [Online]. Available: https://doi.org/10.1016/S0021-9258(17)42076-X
Zhang YB, Howitt J, McCorkle S, Lawrence P, Springer K, Freimuth P. ‘Protein aggregation during overexpression limited by peptide extensions with large net negative charge’, Protein Expr Purif, vol. 36, no. 2, pp. 207–216, Aug. 2004, https://doi.org/10.1016/j.pep.2004.04.020
Köppl C, et al. Fusion Tag Design influences Soluble recombinant protein production in Escherichia coli. Int J Mol Sci. Jul. 2022;23(14). https://doi.org/10.3390/ijms23147678 .
O’Shannessy DJ, O’Donnell KC, Martin J, Brigham-Burke M. ‘Detection and quantitation of hexa-histidine-tagged recombinant proteins on western blots and by a surface plasmon resonance biosensor technique’, Analytical Biochemistry, vol. 229, pp. 119–124, Jul. 1195, https://doi.org/10.1006/abio.1995.1387
Mason B, et al. Expression, purification, and characterization of recombinant nonglycosylated human serum transferrin containing a C-terminal hexahistidine tag. Protein Expr Purif. 2001;23(1):142–50. https://doi.org/10.1006/prep.2001.1480 .
doi: 10.1006/prep.2001.1480
pubmed: 11570856
Summers DK. Sherratt1, ‘Resolution of ColE 1 dimers requires a DNA sequence implicated in the three-dimensional organization of the cer site’. EMBO J. 1988;7(3):851–8.
doi: 10.1002/j.1460-2075.1988.tb02884.x
pubmed: 3294000
pmcid: 454402
Kastenhofer J, Rettenbacher L, Feuchtenhofer L, Mairhofer J, Spadiut O. Inhibition of E. Coli host RNA polymerase allows efficient extracellular recombinant protein production by enhancing outer membrane leakiness. Biotechnol J. Mar. 2021;16(3). https://doi.org/10.1002/biot.202000274 .
Ho NHE, et al. Heterologous expression and characterization of Aquabacterium parvum lipase, a close relative of Ideonella sakaiensis PETase in Escherichia coli. Biochem Eng J. Aug. 2023;197. https://doi.org/10.1016/j.bej.2023.108985 .
Papaneophytou CP. G. Kontopidis 2014 Statistical approaches to maximize recombinant protein expression in Escherichia coli: a general review. Acad Press Inc https://doi.org/10.1016/j.pep.2013.10.016 .
doi: 10.1016/j.pep.2013.10.016
Ravitchandirane G, Bandhu S, Chaudhuri TK. Multimodal approaches for the improvement of the cellular folding of a recombinant iron regulatory protein in E. Coli. Microb Cell Fact. Dec. 2022;21(1). https://doi.org/10.1186/s12934-022-01749-w .
Messens J, Collet JF. ‘Pathways of disulfide bond formation in Escherichia coli’, 2006. https://doi.org/10.1016/j.biocel.2005.12.011
Kadokura H, Katzen F, Beckwith J. ‘Protein Disulfide bond Formation Prokaryotes’. 2003. https://doi.org/10.1146/annurev.biochem.72.121801.161459 .
doi: 10.1146/annurev.biochem.72.121801.161459
Yoshida S et al. Mar., ‘A bacterium that degrades and assimilates poly(ethylene terephthalate)’, Science (1979), 2016, https://doi.org/10.1126/science.aad6359