TY - JOUR
T1 - Selective Synthesis of the Human Drug Metabolite 5′-Hydroxypropranolol by an Evolved Self-Sufficient Peroxygenase
AU - Gomez De Santos, Patricia
AU - Cañellas, Marina
AU - Tieves, Florian
AU - Younes, Sabry H.H.
AU - Molina-Espeja, Patricia
AU - Hofrichter, Martin
AU - Hollmann, Frank
AU - Guallar, Victor
AU - Alcalde, Miguel
PY - 2018
Y1 - 2018
N2 - Propranolol is a widely used beta-blocker that is metabolized by human liver P450 monooxygenases into equipotent hydroxylated human drug metabolites (HDMs). It is paramount for the pharmaceutical industry to evaluate the toxicity and activity of these metabolites, but unfortunately, their synthesis has hitherto involved the use of severe conditions, with poor reaction yields and unwanted byproducts. Unspecific peroxygenases (UPOs) catalyze the selective oxyfunctionalization of C-H bonds, and they are of particular interest in synthetic organic chemistry. Here, we describe the engineering of UPO from Agrocybe aegerita for the efficient synthesis of 5′-hydroxypropranolol (5′-OHP). We employed a structure-guided evolution approach combined with computational analysis, with the aim of avoiding unwanted phenoxyl radical coupling without having to dope the reaction with radical scavengers. The evolved biocatalyst showed a catalytic efficiency enhanced by 2 orders of magnitude and 99% regioselectivity for the synthesis of 5′-OHP. When the UPO mutant was combined with an H2O2 in situ generation system using methanol as sacrificial electron donor, total turnover numbers of up to 264 000 were achieved, offering a cost-effective and readily scalable method to rapidly prepare 5′-OHP.
AB - Propranolol is a widely used beta-blocker that is metabolized by human liver P450 monooxygenases into equipotent hydroxylated human drug metabolites (HDMs). It is paramount for the pharmaceutical industry to evaluate the toxicity and activity of these metabolites, but unfortunately, their synthesis has hitherto involved the use of severe conditions, with poor reaction yields and unwanted byproducts. Unspecific peroxygenases (UPOs) catalyze the selective oxyfunctionalization of C-H bonds, and they are of particular interest in synthetic organic chemistry. Here, we describe the engineering of UPO from Agrocybe aegerita for the efficient synthesis of 5′-hydroxypropranolol (5′-OHP). We employed a structure-guided evolution approach combined with computational analysis, with the aim of avoiding unwanted phenoxyl radical coupling without having to dope the reaction with radical scavengers. The evolved biocatalyst showed a catalytic efficiency enhanced by 2 orders of magnitude and 99% regioselectivity for the synthesis of 5′-OHP. When the UPO mutant was combined with an H2O2 in situ generation system using methanol as sacrificial electron donor, total turnover numbers of up to 264 000 were achieved, offering a cost-effective and readily scalable method to rapidly prepare 5′-OHP.
KW - 5′-hydroxypropranolol
KW - directed evolution
KW - human drug metabolites
KW - in situ HO generation system
KW - peroxidative activity
KW - peroxygenative activity
KW - unspecific peroxygenase
UR - http://resolver.tudelft.nl/uuid:e86ba068-ffac-446b-b14f-b1aaa3ca94f7
UR - http://www.scopus.com/inward/record.url?scp=85046339954&partnerID=8YFLogxK
U2 - 10.1021/acscatal.8b01004
DO - 10.1021/acscatal.8b01004
M3 - Article
SN - 2155-5435
VL - 8
SP - 4789
EP - 4799
JO - ACS Catalysis
JF - ACS Catalysis
IS - 6
ER -