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Hendil-Forssell, PeterORCID iD iconorcid.org/0000-0001-9001-9271
Publications (5 of 5) Show all publications
Finnveden, M., Hendil-Forssell, P., Claudino, M., Johansson, M. & Martinelle, M. (2019). Lipase-Catalyzed Synthesis of Renewable Plant Oil-Based Polyamides.. Polymers, 11(11), Article ID 1730.
Open this publication in new window or tab >>Lipase-Catalyzed Synthesis of Renewable Plant Oil-Based Polyamides.
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2019 (English)In: Polymers, ISSN 2073-4360, E-ISSN 2073-4360, Vol. 11, no 11, article id 1730Article in journal (Refereed) Published
Abstract [en]

Enzyme catalyzed synthesis of renewable polyamides was investigated using Candida antarctica lipase B. A fatty acid-derived AB-type functional monomer, having one amine and one methyl ester functionality, was homopolymerized at 80 and 140 °C. Additionally, the organobase 1,5,7-triazabicyclo[4.4.0]dec-5-ene (TBD) was used as a catalyst. The results from the two catalysts were comparable. However, the amount of lipase added was 1.2 × 103 times lower, showing that the lipase was a more efficient catalyst for this system as compared to TBD. Moreover, the AB-type monomer was copolymerized with 1,12-diaminododecane to synthesize oligoamides of two different lengths.

Place, publisher, year, edition, pages
MDPI, 2019
Keywords
Candida antarctica lipase B, bio-based polyamides, enzymatic polymerization
National Category
Biocatalysis and Enzyme Technology
Research subject
Biotechnology
Identifiers
urn:nbn:se:kth:diva-264924 (URN)10.3390/polym11111730 (DOI)000503279200003 ()31652736 (PubMedID)2-s2.0-85075579461 (Scopus ID)
Funder
Swedish Research Council Formas, 211-2013-70EU, FP7, Seventh Framework Programme, 266025
Note

QC 20191205

Available from: 2019-12-05 Created: 2019-12-05 Last updated: 2020-01-10Bibliographically approved
Hendil-Forssell, P. (2016). Rational engineering of esterases for improved amidase specificity in amide synthesis and hydrolysis. (Doctoral dissertation). Stockholm: KTH Royal Institute of Technology
Open this publication in new window or tab >>Rational engineering of esterases for improved amidase specificity in amide synthesis and hydrolysis
2016 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Biocatalysis is an ever evolving field that uses enzymes or microorganisms for chemical synthesis. By utilizing enzymes that generally have evolved for specific reactions under mild conditions and temperatures, biocatalysis can be a more environmentally friendly option compared to traditional chemistry.

Amide-type chemistries are important and bond formation avoiding poor atom economy is of high priority in organic chemistry. Biocatalysis could potentially be a solution but restricted substrate scope is a limitation. Esterases/lipases usually display broad substrate scope and catalytic promiscuity but are poor at hydrolyzing amides compared to amidases/proteases. The difference between the two enzyme classes is hypothesized to reside in one key hydrogen bond present in amidases, which facilitates the transition state for nitrogen inversion during catalysis.

In this thesis the work has been focused on introducing a stabilizing hydrogen bond acceptor in esterases, mimicking that found in amidases, to develop better enzymatic catalysts for amide-based chemistries.

By two strategies, side-chain or water interaction, variants were created in three esterases that displayed up to 210-times increased relative amidase specificity compared to the wild type. The best variant displayed reduced activation enthalpy corresponding to a weak hydrogen bond. The results show an estimated lower limit on how much the hydrogen bond can be worth to catalysis.

MsAcT catalyze kinetically controlled N-acylations in water. An enzymatic one-pot one-step cascade was developed for the formation of amides from aldehydes in water that gave 97% conversion. In addition, engineered variants of MsAcT with increased substrate scope could synthesize an amide in water with 81% conversion, where the wild type gave no conversion. Moreover, variants of MsAcT displayed up to 32-fold change in specificity towards amide synthesis and a switch in reaction preference favoring amide over ester synthesis.

Place, publisher, year, edition, pages
Stockholm: KTH Royal Institute of Technology, 2016. p. 76
Series
TRITA-BIO-Report, ISSN 1654-2312 ; 2016:21
Keywords
Amidase, Biocatalysis, Enzyme, Esterase, Enzyme engineering, Lipase, Substrate specificity
National Category
Biocatalysis and Enzyme Technology
Research subject
Biotechnology
Identifiers
urn:nbn:se:kth:diva-196892 (URN)978-91-7729-210-4 (ISBN)
Public defence
2016-12-16, FD5, AlbaNova University Center, Roslagstullsbacken 21, Stockholm, 10:00 (English)
Opponent
Supervisors
Note

QC 20161125

Available from: 2016-11-25 Created: 2016-11-24 Last updated: 2016-11-25Bibliographically approved
Hendil-Forssell, P., Semlitsch, S. & Martinelle, M.Engineering the esterase/acyltransferase from Mycobacterium smegmatis: extended substrate scope for amide synthesis in water.
Open this publication in new window or tab >>Engineering the esterase/acyltransferase from Mycobacterium smegmatis: extended substrate scope for amide synthesis in water
(English)Manuscript (preprint) (Other academic)
Abstract [en]

Some esterases/lipases display high acyl transfer activity, favoring alcoholysis over hydrolysis, which make them valuable catalysts for synthesis reactions in aqueous media. An esterase from Mycobacterium smegmatis, MsAcT, has been characterized as an efficient catalyst for ester synthesis in water. The acyl donor specificity for MsAcT was however found to be very narrow and the enzyme displayed no activity towards esters with larger acyl group than butyrate. With rational engineering, the narrow acyl donor specificity of wild type MsAcT enzyme was altered and variants displaying extended substrate scope were generated. A double mutant, T93A/F154A, could accommodate methyl nonanoate as substrate, i.e. five carbons longer acyl group as compared to wild type, without compromising the acyl transfer capabilities. With similar selectivity towards a broad range of acyl donors (propionate to nonanoate) this is a more applicable catalyst than the wild type. Furthermore, the T93A/F154A variant was an efficient catalyst for synthesis of N-benzylhexanamide in water using methyl hexanoate as acyl donor, which is not a substrate for the wild type enzyme. The conversion reached 81% and the enzyme variant could potentially be used to produce amides in water with a wide variety of acyl donors.

National Category
Biocatalysis and Enzyme Technology
Research subject
Biotechnology
Identifiers
urn:nbn:se:kth:diva-196890 (URN)
Note

QC 20161129

Available from: 2016-11-24 Created: 2016-11-24 Last updated: 2017-08-23Bibliographically approved
Finnveden, M., Hendil-Forssell, P., Claudino, M., Johansson, M. & Martinelle, M.Lipase Catalyzed Synthesis of renewable plant oil-based polyamides.
Open this publication in new window or tab >>Lipase Catalyzed Synthesis of renewable plant oil-based polyamides
Show others...
(English)Manuscript (preprint) (Other academic)
Abstract [en]

Enzyme catalyzed synthesis towards renewable polyamides was investigated using Candida antarctica lipase B. A fatty acid-derived AB-type functional monomer, having one amine and one methyl ester functionality was homopolymerized at 80 and 140°C. Additionally, the organobase 1,5,7-triazabicyclo[4.4.0]dec-5-ene (TBD) was used as catalyst. The results from the two catalysts were comparable. However, the amount of lipase added was 1200 times lower showing that the lipase was a more efficient catalyst for this system as compared to TBD. Moreover, the AB type monomer was copolymerized with 1,12-diaminododecan to synthesize oligoamides of two different lengths.

Keywords
Candida antarctica lipase B; bio-based polyamides; enzymatic polymerization
National Category
Biocatalysis and Enzyme Technology
Research subject
Biotechnology; Fibre and Polymer Science
Identifiers
urn:nbn:se:kth:diva-257676 (URN)
Note

QCR 20190903

Available from: 2019-09-02 Created: 2019-09-02 Last updated: 2019-09-04Bibliographically approved
Hendil-Forssell, P., Semlitsch, S. & Martinelle, M.Rational engineering of an esterase/acyltransferase for improved amidase specificity in amide synthesis and hydrolysis.
Open this publication in new window or tab >>Rational engineering of an esterase/acyltransferase for improved amidase specificity in amide synthesis and hydrolysis
(English)Manuscript (preprint) (Other academic)
Abstract [en]

The esterase/acyltransferase from Mycobacterium smegmatis, MsAcT, display high acyltransfer capacity in water media with demonstrations found for both ester and amide syntheses. However, it has recently been discovered that esterases in contrast to amidases lack a key hydrogen bond in the transition state, donated by the scissile NH-group of the substrate. Esterases with improved amidase performance have been achieved with the introduction of amino-acid side chains or water network as hydrogen bond acceptors. Using the esterase from Mycobacterium smegmatis, MsAcT, the influence of this hydrogen bond was studied in both amide hydrolysis and synthesis, using a rational engineering approach. Two positions were selected for mutagenesis and enzyme variants with improved performance in amide synthesis and hydrolysis were generated. Compared to the wild-type, variant F154A had the highest absolute increase in amidase specificity (11-fold) and I194Q had the greatest change in relative amidase versus esterase reaction specificity (160-fold). The relative reaction specificities for amide over ester synthesis followed a similar trend as that of hydrolysis and the best variant was I194Q with a 32-fold increase compared to wt. Based on MD-simulations water seems to play an important role in the transition state as a hydrogen bond bridge between the NH-group of the amide substrate and the enzyme.

National Category
Biocatalysis and Enzyme Technology
Research subject
Biotechnology
Identifiers
urn:nbn:se:kth:diva-196891 (URN)
Note

QC 20161129

Available from: 2016-11-24 Created: 2016-11-24 Last updated: 2016-11-29Bibliographically approved
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ORCID iD: ORCID iD iconorcid.org/0000-0001-9001-9271

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