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Publications (10 of 36) Show all publications
Farhat, W., Stamm, A., Robert-Monpate, M., Biundo, A. & Syrén, P.-O. (2019). Biocatalysis for terpene-based polymers. Zeitschrift für Naturforschung C - A Journal of Biosciences, 74(3-4), 90-99
Open this publication in new window or tab >>Biocatalysis for terpene-based polymers
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2019 (English)In: Zeitschrift für Naturforschung C - A Journal of Biosciences, ISSN 0939-5075, E-ISSN 1865-7125, Vol. 74, no 3-4, p. 90-99Article in journal (Refereed) Published
Abstract [en]

Accelerated generation of bio-based materials is vital to replace current synthetic polymers obtained from petroleum with more sustainable options. However, many building blocks available from renewable resources mainly contain unreactive carbon-carbon bonds, which obstructs their efficient polymerization. Herein, we highlight the potential of applying biocatalysis to afford tailored functionalization of the inert carbocyclic core of multicyclic terpenes toward advanced materials. As a showcase, we unlock the inherent monomer reactivity of norcamphor, a bicyclic ketone used as a monoterpene model system in this study, to afford polyesters with unprecedented backbones. The efficiencies of the chemical and enzymatic Baeyer-Villiger transformation in generating key lactone intermediates are compared. The concepts discussed herein are widely applicable for the valorization of terpenes and other cyclic building blocks using chemoenzymatic strategies.

Place, publisher, year, edition, pages
WALTER DE GRUYTER GMBH, 2019
Keywords
biocatalysis, biopolymers, oxidoreductases, polyesters, terpenes
National Category
Polymer Chemistry
Identifiers
urn:nbn:se:kth:diva-246249 (URN)10.1515/znc-2018-0199 (DOI)000459670900006 ()30789828 (PubMedID)2-s2.0-85061735936 (Scopus ID)
Note

QC 20190402

Available from: 2019-04-02 Created: 2019-04-02 Last updated: 2019-04-04Bibliographically approved
Stamm, A., Tengdelius, M., Schmidt, B., Engström, J., Syrén, P.-O., Fogelström, L. & Malmström, E. (2019). Chemo- enzymatic pathways toward pinene- based renewable materials. Green Chemistry, 21(10), 2720-2731
Open this publication in new window or tab >>Chemo- enzymatic pathways toward pinene- based renewable materials
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2019 (English)In: Green Chemistry, ISSN 1463-9262, E-ISSN 1463-9270, Vol. 21, no 10, p. 2720-2731Article in journal (Refereed) Published
Abstract [en]

Sobrerol methacrylate (SobMA) was synthesized and subsequently polymerized using different chemical and enzymatic routes. Sobrerol was enzymatically converted from -pinene in a small model scale by a Cytochrome P450 mutant from Bacillus megaterium. Conversion of sobrerol into SobMA was performed using both classical ester synthesis, i.e., acid chloride-reactions in organic solvents, and a more green approach, the benign lipase catalysis. Sobrerol was successfully esterified, leaving the tertiary alcohol and ene to be used for further chemistry. SobMA was polymerized into PSobMA using different radical polymerization techniques, including free radical (FR), controlled procedures (Reversible Addition Fragmentation chain-Transfer polymerization, (RAFT) and Atom Transfer Radical Polymerization (ATRP)) as well as by enzyme catalysis (horseradish peroxidase-mediated free radical polymerization). The resulting polymers showed high glass-transition temperatures (T-g) around 150 degrees C, and a thermal degradation onset above 200 degrees C. It was demonstrated that the T-g could be tailored by copolymerizing SobMa with appropriate methacrylate monomers and that the Flory-Fox equation could be used to predict the T-g. The versatility of PSobMA was further demonstrated by forming crosslinked thin films, either using the ene'-functionality for photochemically initiated thiol-ene'-chemistry, or reacting the tertiary hydroxyl-group with hexamethoxymethylmelamine, as readily used for thermally curing coatings systems.

National Category
Polymer Technologies Polymer Chemistry
Identifiers
urn:nbn:se:kth:diva-252972 (URN)10.1039/c9gc00718k (DOI)000468627800016 ()2-s2.0-85066853137 (Scopus ID)
Note

QC 20190812

Available from: 2019-08-12 Created: 2019-08-12 Last updated: 2019-08-12Bibliographically approved
Fogelström, L., Stamm, A., Tengdelius, M., Syrén, P.-O. & Malmström, E. (2019). New chemo-enzymatic pathways for sustainable terpene-based polymeric materials. Paper presented at National Meeting of the American-Chemical-Society (ACS), MAR 31-APR 04, 2019, Orlando, FL. Abstracts of Papers of the American Chemical Society, 257
Open this publication in new window or tab >>New chemo-enzymatic pathways for sustainable terpene-based polymeric materials
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2019 (English)In: Abstracts of Papers of the American Chemical Society, ISSN 0065-7727, Vol. 257Article in journal, Meeting abstract (Other academic) Published
Place, publisher, year, edition, pages
AMER CHEMICAL SOC, 2019
National Category
Paper, Pulp and Fiber Technology
Identifiers
urn:nbn:se:kth:diva-257595 (URN)000478860502626 ()
Conference
National Meeting of the American-Chemical-Society (ACS), MAR 31-APR 04, 2019, Orlando, FL
Note

QC 20190919

Available from: 2019-09-19 Created: 2019-09-19 Last updated: 2019-09-19Bibliographically approved
Malmström, E., Fogelström, L., Stamm, A., Tengdelius, M., Biundo, A. & Syrén, P.-O. (2019). Sustainable terpene-based polymeric materials. Paper presented at 257th National Meeting of the American-Chemical-Society (ACS), MAR 31-APR 04, 2019, Orlando, FL. Abstracts of Papers of the American Chemical Society, 257
Open this publication in new window or tab >>Sustainable terpene-based polymeric materials
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2019 (English)In: Abstracts of Papers of the American Chemical Society, ISSN 0065-7727, Vol. 257Article in journal, Meeting abstract (Other academic) Published
Place, publisher, year, edition, pages
American Chemical Society (ACS), 2019
National Category
Other Chemistry Topics
Identifiers
urn:nbn:se:kth:diva-257660 (URN)000478861204618 ()
Conference
257th National Meeting of the American-Chemical-Society (ACS), MAR 31-APR 04, 2019, Orlando, FL
Note

QC 20190903

Available from: 2019-09-03 Created: 2019-09-03 Last updated: 2019-09-03Bibliographically approved
Hendrikse, N., Charpentier, G., Nordling, E. & Syrén, P.-O. (2018). Ancestral diterpene cyclases show increased thermostability and substrate acceptance. The FEBS Journal, 285(24), 4660-4673
Open this publication in new window or tab >>Ancestral diterpene cyclases show increased thermostability and substrate acceptance
2018 (English)In: The FEBS Journal, ISSN 1742-464X, E-ISSN 1742-4658, Vol. 285, no 24, p. 4660-4673Article in journal (Refereed) Published
Abstract [en]

Bacterial diterpene cyclases are receiving increasing attention in biocatalysis and synthetic biology for the sustainable generation of complex multicyclic building blocks. Herein, we explore the potential of ancestral sequence reconstruction (ASR) to generate remodeled cyclases with enhanced stability, activity, and promiscuity. Putative ancestors of spiroviolene synthase, a bacterial class I diterpene cyclase, display an increased yield of soluble protein of up to fourfold upon expression in the model organism Escherichia coli. Two of the resurrected enzymes, with an estimated age of approximately 1.7 million years, display an upward shift in thermostability of 7-13 degrees C. Ancestral spiroviolene synthases catalyze cyclization of the natural C-20-substrate geranylgeranyl diphosphate (GGPP) and also accept C-15 farnesyl diphosphate (FPP), which is not converted by the extant enzyme. In contrast, the consensus sequence generated from the corresponding multiple sequence alignment was found to be inactive toward both substrates. Mutation of a nonconserved position within the aspartate-rich motif of the reconstructed ancestral cyclases was associated with modest effects on activity and relative substrate specificity (i.e., k(cat)/K-M for GGPP over k(cat)/K-M for FPP). Kinetic analyses performed at different temperatures reveal a loss of substrate saturation, when going from the ancestor with highest thermostability to the modern enzyme. The kinetics data also illustrate how an increase in temperature optimum of biocatalysis is reflected in altered entropy and enthalpy of activation. Our findings further highlight the potential and limitations of applying ASR to biosynthetic machineries in secondary metabolism.

Place, publisher, year, edition, pages
Wiley-VCH Verlagsgesellschaft, 2018
Keywords
ancestral sequence reconstruction, diterpene cyclase, spiroviolene synthase, protein stability, promiscuity
National Category
Chemical Sciences
Identifiers
urn:nbn:se:kth:diva-240741 (URN)10.1111/febs.14686 (DOI)000453570200010 ()30369053 (PubMedID)2-s2.0-85056620593 (Scopus ID)
Funder
VINNOVA, 2016-03344Swedish Foundation for Strategic Research , ID16-0036Science for Life Laboratory - a national resource center for high-throughput molecular bioscience
Note

QC 20190103

Available from: 2019-01-03 Created: 2019-01-03 Last updated: 2019-01-07Bibliographically approved
Pavlidis, I. V., Hendrikse, N. & Syrén, P.-O. (2018). Chapter 5: Computational Techniques for Efficient Biocatalysis. RSC Catalysis Series (32), 119-152
Open this publication in new window or tab >>Chapter 5: Computational Techniques for Efficient Biocatalysis
2018 (English)In: RSC Catalysis Series, no 32, p. 119-152Article in journal (Refereed) Published
Abstract [en]

Addressing some of the most challenging problems that we face today, including depletion of natural resources, sustainable energy production and the generation of green polymeric materials by the biocatalytic upcycling of renewable synthons, requires an expansion of the current available biochemical reaction space. Creating biocatalysts harboring novel chemistries - whether inside or outside the cell - is dependent on the discovery of novel enzymes and metabolic pathways, together with the de novo design of enzymes and directed evolution. Herein we review the high potential of using bioinformatics and in silico computer modelling tools to guide protein engineering and to enhance our fundamental understanding of biocatalysis. Following an overview of technical considerations and the current state-of-the art in sequence- and structure-based protein engineering methodologies, we highlight recent successful examples of their implementation in biocatalysis and synthetic biology. Moreover, we discuss how selected computational tools in concert with experimental biocatalysis could decipher how the sequence, structure and dynamics of proteins dictate their function. Using the methodologies discussed in this chapter, an accelerated biocatalytic manufacturing of chemicals, pharmaceuticals, biofuels and monomeric building blocks is envisioned.

Place, publisher, year, edition, pages
Royal Society of Chemistry, 2018
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:kth:diva-236399 (URN)10.1039/9781788010450-00117 (DOI)2-s2.0-85049330104 (Scopus ID)
Note

QC 20181101

Available from: 2018-11-01 Created: 2018-11-01 Last updated: 2018-11-01Bibliographically approved
Syrén, P.-O. (2018). Enzymatic Hydrolysis of Tertiary Amide Bonds by anti Nucleophilic Attack and Protonation. Journal of Organic Chemistry, 83(21), 13543-13548
Open this publication in new window or tab >>Enzymatic Hydrolysis of Tertiary Amide Bonds by anti Nucleophilic Attack and Protonation
2018 (English)In: Journal of Organic Chemistry, ISSN 0022-3263, E-ISSN 1520-6904, Vol. 83, no 21, p. 13543-13548Article in journal (Refereed) Published
Abstract [en]

The molecular mechanisms conferring high resistance of planar tertiary amide bonds to hydrolysis by most enzymes have remained elusive. To provide a chemical explanation to this unresolved puzzle, UB3LYP calculations were performed on an active site model of Xaa-Pro peptidases. The calculated reaction mechanism demonstrates that biocatalysts capable of tertiary amide bond hydrolysis capitalize on anti nucleophilic attack and protonation of the amide nitrogen, in contrast to the traditional syn displayed by amidases and proteases acting on secondary amide bonds.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2018
National Category
Chemical Sciences
Identifiers
urn:nbn:se:kth:diva-239484 (URN)10.1021/acs.joc.8b02053 (DOI)000449443200058 ()30256635 (PubMedID)2-s2.0-85054958555 (Scopus ID)
Funder
Swedish Research Council Formas, 2017-01116 2017-01116
Note

QC 20181127

Available from: 2018-11-27 Created: 2018-11-27 Last updated: 2019-08-20Bibliographically approved
Gustafsson, C., Vassiliev, S., Kürten, C., Syrén, P.-O. & Brinck, T. (2017). MD Simulations Reveal Complex Water Paths in Squalene–Hopene Cyclase: Tunnel-Obstructing Mutations Increase the Flow of Water in the Active Site. ACS Omega, 2(11), 8495-8506
Open this publication in new window or tab >>MD Simulations Reveal Complex Water Paths in Squalene–Hopene Cyclase: Tunnel-Obstructing Mutations Increase the Flow of Water in the Active Site
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2017 (English)In: ACS Omega, ISSN 2470-1343, Vol. 2, no 11, p. 8495-8506Article in journal (Refereed) Published
Abstract [en]

Squalene–hopene cyclase catalyzes the cyclization of squalene to hopanoids. A previous study has identified a network of tunnels in the protein, where water molecules have been indicated to move. Blocking these tunnels by site-directed mutagenesis was found to change the activation entropy of the catalytic reaction from positive to negative with a concomitant lowering of the activation enthalpy. As a consequence, some variants are faster and others are slower than the wild type (wt) in vitro under optimal reaction conditions for the wt. In this study, molecular dynamics (MD) simulations have been performed for the wt and the variants to investigate how the mutations affect the protein structure and the water flow in the enzyme, hypothetically influencing the activation parameters. Interestingly, the tunnel-obstructing variants are associated with an increased flow of water in the active site, particularly close to the catalytic residue Asp376. MD simulations with the substrate present in the active site indicate that the distance for the rate-determining proton transfer between Asp376 and the substrate is longer in the tunnel-obstructing protein variants than in the wt. On the basis of the previous experimental results and the current MD results, we propose that the tunnel-obstructing variants, at least partly, could operate by a different catalytic mechanism, where the proton transfer may have contributions from a Grotthuss-like mechanism.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2017
National Category
Biocatalysis and Enzyme Technology Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:kth:diva-234939 (URN)10.1021/acsomega.7b01084 (DOI)000418744100113 ()
Funder
Science for Life Laboratory - a national resource center for high-throughput molecular bioscience
Note

QC 20180914

Available from: 2018-09-13 Created: 2018-09-13 Last updated: 2018-09-18Bibliographically approved
Eriksson, A., Kürten, C. & Syrén, P.-O. (2017). Protonation-Initiated Cyclization by a ClassII Terpene Cyclase Assisted by Tunneling. ChemBioChem (Print), 18(23), 2301-2305
Open this publication in new window or tab >>Protonation-Initiated Cyclization by a ClassII Terpene Cyclase Assisted by Tunneling
2017 (English)In: ChemBioChem (Print), ISSN 1439-4227, E-ISSN 1439-7633, Vol. 18, no 23, p. 2301-2305Article in journal (Refereed) Published
Abstract [en]

Terpenes represent one of the most diversified classes of natural products with potent biological activities. The key to the myriad of polycyclic terpene skeletons with crucial functions in organisms from all kingdoms of life are terpene cyclase enzymes. These biocatalysts enable stereospecific cyclization of relatively simple, linear, prefolded polyisoprenes by highly complex, partially concerted, electrophilic cyclization cascades that remain incompletely understood. Herein, additional mechanistic light is shed on terpene biosynthesis by kinetic studies in mixed H2O/D2O buffers of a classII bacterial ent-copalyl diphosphate synthase. Mass spectrometry determination of the extent of deuterium incorporation in the bicyclic product, reminiscent of initial carbocation formation by protonation, resulted in a large kinetic isotope effect of up to seven. Kinetic analysis at different temperatures confirmed that the isotope effect was independent of temperature, which is consistent with hydrogen tunneling.

Place, publisher, year, edition, pages
Wiley-VCH Verlagsgesellschaft, 2017
Keywords
biosynthesis, enzyme catalysis, isotope effects, kinetics, reaction mechanisms
National Category
Other Chemical Engineering
Identifiers
urn:nbn:se:kth:diva-220459 (URN)10.1002/cbic.201700443 (DOI)000417219500006 ()
Funder
Swedish Research Council, 621-2013-5138AFA Insurance, 17-359Science for Life Laboratory - a national resource center for high-throughput molecular bioscience
Note

QC 20180104

Available from: 2018-01-04 Created: 2018-01-04 Last updated: 2018-09-13Bibliographically approved
Hendrikse, N. M., Stromberg, P., Nordling, E. & Syrén, P.-O. (2017). Redesign of biosynthetic enzymes using ancestral sequence reconstruction. Paper presented at 42nd Congress of the Federation-of-European-Biochemical-Societies (FEBS) on From Molecules to Cells and Back, SEP 10-14, 2017, Jerusalem, ISRAEL. The FEBS Journal, 284, 86-87
Open this publication in new window or tab >>Redesign of biosynthetic enzymes using ancestral sequence reconstruction
2017 (English)In: The FEBS Journal, ISSN 1742-464X, E-ISSN 1742-4658, Vol. 284, p. 86-87Article in journal (Refereed) Published
Place, publisher, year, edition, pages
Wiley-Blackwell, 2017
National Category
Physical Chemistry
Identifiers
urn:nbn:se:kth:diva-216635 (URN)000409918901053 ()
Conference
42nd Congress of the Federation-of-European-Biochemical-Societies (FEBS) on From Molecules to Cells and Back, SEP 10-14, 2017, Jerusalem, ISRAEL
Note

QC 20171101

Available from: 2017-11-01 Created: 2017-11-01 Last updated: 2017-11-01Bibliographically approved
Organisations
Identifiers
ORCID iD: ORCID iD iconorcid.org/0000-0002-4066-2776

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