Abstract
Hydroxynitrile lyase from the white rabbit's foot fern Davallia tyermannii (DtHNL) catalyzes the enantioselective synthesis of α-cyanohydrins, which are key building blocks for pharmaceutical and agrochemical industries. An efficient and competitive process necessitates the availability and robustness of the biocatalyst. Herein, the recombinant production of DtHNL1 in Komagataella phaffii, yielding approximately 900 000 U L-1 , is described. DtHNL1 constitutes approximately 80 % of the total protein content. The crude enzyme was immobilized. Crosslinked enzyme aggregates (CLEAs) resulted in significant enhancement of the biocatalyst stability under acidic conditions (activity retained after 168 h at pH 2.4). The DtHNL1-CLEA was employed for (R)-mandelonitrile synthesis (99 % conversion, 98 % enantiomeric excess) in a biphasic system, and evaluated for the synthesis of (R)-hydroxypivaldehyde cyanohydrin under reaction conditions that immediately inactivated non-immobilized DtHNL1. The results show the DtHNL1-CLEA to be a stable biocatalyst for the synthesis of enantiomerically pure cyanohydrins under acidic conditions.
| Originalsprache | englisch |
|---|---|
| Aufsatznummer | 10.1002/cbic.201700419 |
| Seiten (von - bis) | 312-316 |
| Fachzeitschrift | ChemBioChem |
| Jahrgang | 19 |
| Ausgabenummer | 4 |
| Frühes Online-Datum | 13 Nov 2017 |
| DOIs | |
| Publikationsstatus | Veröffentlicht - 2018 |
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Fields of Expertise
- Human- & Biotechnology
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Production of Hydroxynitrile Lyase from Davallia tyermannii (DtHNL) in Komagataella phaffii and Its Immobilization as a CLEA to Generate a Robust Biocatalyst. / Lanfranchi, Elisa; Grill, Birgit; Raghoebar, Zainab; Van Pelt, Sander; Sheldon, Roger A; Steiner, Kerstin; Glieder, Anton; Winkler, Margit.
in: ChemBioChem, Jahrgang 19, Nr. 4, 10.1002/cbic.201700419, 2018, S. 312-316.Publikation: Beitrag in einer Fachzeitschrift › Artikel › Forschung › Begutachtung
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TY - JOUR
T1 - Production of Hydroxynitrile Lyase from Davallia tyermannii (DtHNL) in Komagataella phaffii and Its Immobilization as a CLEA to Generate a Robust Biocatalyst
AU - Lanfranchi, Elisa
AU - Grill, Birgit
AU - Raghoebar, Zainab
AU - Van Pelt, Sander
AU - Sheldon, Roger A
AU - Steiner, Kerstin
AU - Glieder, Anton
AU - Winkler, Margit
N1 - © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
PY - 2018
Y1 - 2018
N2 - Hydroxynitrile lyase from the white rabbit's foot fern Davallia tyermannii (DtHNL) catalyzes the enantioselective synthesis of α-cyanohydrins, which are key building blocks for pharmaceutical and agrochemical industries. An efficient and competitive process necessitates the availability and robustness of the biocatalyst. Herein, the recombinant production of DtHNL1 in Komagataella phaffii, yielding approximately 900 000 U L-1 , is described. DtHNL1 constitutes approximately 80 % of the total protein content. The crude enzyme was immobilized. Crosslinked enzyme aggregates (CLEAs) resulted in significant enhancement of the biocatalyst stability under acidic conditions (activity retained after 168 h at pH 2.4). The DtHNL1-CLEA was employed for (R)-mandelonitrile synthesis (99 % conversion, 98 % enantiomeric excess) in a biphasic system, and evaluated for the synthesis of (R)-hydroxypivaldehyde cyanohydrin under reaction conditions that immediately inactivated non-immobilized DtHNL1. The results show the DtHNL1-CLEA to be a stable biocatalyst for the synthesis of enantiomerically pure cyanohydrins under acidic conditions.
AB - Hydroxynitrile lyase from the white rabbit's foot fern Davallia tyermannii (DtHNL) catalyzes the enantioselective synthesis of α-cyanohydrins, which are key building blocks for pharmaceutical and agrochemical industries. An efficient and competitive process necessitates the availability and robustness of the biocatalyst. Herein, the recombinant production of DtHNL1 in Komagataella phaffii, yielding approximately 900 000 U L-1 , is described. DtHNL1 constitutes approximately 80 % of the total protein content. The crude enzyme was immobilized. Crosslinked enzyme aggregates (CLEAs) resulted in significant enhancement of the biocatalyst stability under acidic conditions (activity retained after 168 h at pH 2.4). The DtHNL1-CLEA was employed for (R)-mandelonitrile synthesis (99 % conversion, 98 % enantiomeric excess) in a biphasic system, and evaluated for the synthesis of (R)-hydroxypivaldehyde cyanohydrin under reaction conditions that immediately inactivated non-immobilized DtHNL1. The results show the DtHNL1-CLEA to be a stable biocatalyst for the synthesis of enantiomerically pure cyanohydrins under acidic conditions.
U2 - 10.1002/cbic.201700419
DO - 10.1002/cbic.201700419
M3 - Article
VL - 19
SP - 312
EP - 316
JO - ChemBioChem
JF - ChemBioChem
SN - 1439-4227
IS - 4
M1 - 10.1002/cbic.201700419
ER -