TY - JOUR
T1 - Laccase did it again
T2 - A scalable and clean regeneration system for NAD+ and its application in the synthesis of 12-oxo-hydroxysteroids
AU - Tonin, Fabio
AU - Martì, Elisabet
AU - Arends, Isabel W.C.E.
AU - Hanefeld, Ulf
PY - 2020
Y1 - 2020
N2 - The specific oxidation of 12α-OH group of hydroxysteroids is required for the preparation of cheno-and ursodeoxycholic acid (CDCA and UDCA, respectively). The C12 oxidation of hydroxysteroids into their 12-oxo derivatives can selectively be performed by employing 12α-hydroxysteroid dehydrogenases. These enzymes use NAD(P)+ as an electron acceptor, which has to be re-oxidized in a so-called “regeneration system”. Recently, the enzyme NAD(P)H oxidase (NOX) was applied for the regeneration of NAD+ in the enzymatic preparation of 12-oxo-CDCA from cholic acid (CA), which allows air to be used as an oxidant. However, the NOX system suffers from low activity and low stability. Moreover, the substrate loading is limited to 10 mM. In this study, the laccase/mediator system was investigated as a possible alternative to NOX, employing air as an oxidant. The laccase/mediator system shows higher productivity and scalability than the NOX system. This was proven with a preparative biotransformation of 20 g of CA into 12-oxo-CDCA (92% isolated yield) by employing a substrate loading of 120 mM (corresponding to 50 g/L). Additionally, the performance of the laccase/mediator system was compared with a classical ADH/acetone regeneration system and with other regeneration systems reported in literature.
AB - The specific oxidation of 12α-OH group of hydroxysteroids is required for the preparation of cheno-and ursodeoxycholic acid (CDCA and UDCA, respectively). The C12 oxidation of hydroxysteroids into their 12-oxo derivatives can selectively be performed by employing 12α-hydroxysteroid dehydrogenases. These enzymes use NAD(P)+ as an electron acceptor, which has to be re-oxidized in a so-called “regeneration system”. Recently, the enzyme NAD(P)H oxidase (NOX) was applied for the regeneration of NAD+ in the enzymatic preparation of 12-oxo-CDCA from cholic acid (CA), which allows air to be used as an oxidant. However, the NOX system suffers from low activity and low stability. Moreover, the substrate loading is limited to 10 mM. In this study, the laccase/mediator system was investigated as a possible alternative to NOX, employing air as an oxidant. The laccase/mediator system shows higher productivity and scalability than the NOX system. This was proven with a preparative biotransformation of 20 g of CA into 12-oxo-CDCA (92% isolated yield) by employing a substrate loading of 120 mM (corresponding to 50 g/L). Additionally, the performance of the laccase/mediator system was compared with a classical ADH/acetone regeneration system and with other regeneration systems reported in literature.
KW - Biocatalysis
KW - Hydroxysteroid dehydrogenase
KW - Laccase
KW - Laccase-mediator
KW - NAD regeneration
UR - http://www.scopus.com/inward/record.url?scp=85086667218&partnerID=8YFLogxK
U2 - 10.3390/catal10060677
DO - 10.3390/catal10060677
M3 - Article
AN - SCOPUS:85086667218
SN - 2073-4344
VL - 10
JO - Catalysts
JF - Catalysts
IS - 6
M1 - 677
ER -