TY - JOUR
T1 - Engineering a Highly Regioselective Fungal Peroxygenase for the Synthesis of Hydroxy Fatty Acids
AU - Gomez de Santos, Patricia
AU - González-Benjumea, Alejandro
AU - Fernandez-Garcia, Angela
AU - Aranda, Carmen
AU - Wu, Yinqi
AU - But, Andrada
AU - Molina-Espeja, Patricia
AU - Zhang, Wuyuan
AU - Hollmann, Frank
AU - More Authors, null
PY - 2023
Y1 - 2023
N2 - The hydroxylation of fatty acids is an appealing reaction in synthetic chemistry, although the lack of selective catalysts hampers its industrial implementation. In this study, we have engineered a highly regioselective fungal peroxygenase for the ω-1 hydroxylation of fatty acids with quenched stepwise over-oxidation. One single mutation near the Phe catalytic tripod narrowed the heme cavity, promoting a dramatic shift toward subterminal hydroxylation with a drop in the over-oxidation activity. While crystallographic soaking experiments and molecular dynamic simulations shed light on this unique oxidation pattern, the selective biocatalyst was produced by Pichia pastoris at 0.4 g L−1 in a fed-batch bioreactor and used in the preparative synthesis of 1.4 g of (ω-1)-hydroxytetradecanoic acid with 95 % regioselectivity and 83 % ee for the S enantiomer.
AB - The hydroxylation of fatty acids is an appealing reaction in synthetic chemistry, although the lack of selective catalysts hampers its industrial implementation. In this study, we have engineered a highly regioselective fungal peroxygenase for the ω-1 hydroxylation of fatty acids with quenched stepwise over-oxidation. One single mutation near the Phe catalytic tripod narrowed the heme cavity, promoting a dramatic shift toward subterminal hydroxylation with a drop in the over-oxidation activity. While crystallographic soaking experiments and molecular dynamic simulations shed light on this unique oxidation pattern, the selective biocatalyst was produced by Pichia pastoris at 0.4 g L−1 in a fed-batch bioreactor and used in the preparative synthesis of 1.4 g of (ω-1)-hydroxytetradecanoic acid with 95 % regioselectivity and 83 % ee for the S enantiomer.
KW - Hydroxy Fatty Acids
KW - Over-Oxidation
KW - Protein Engineering
KW - Regioselectivity
KW - Unspecific Peroxygenase
UR - http://www.scopus.com/inward/record.url?scp=85146709046&partnerID=8YFLogxK
U2 - 10.1002/anie.202217372
DO - 10.1002/anie.202217372
M3 - Article
C2 - 36583658
AN - SCOPUS:85146709046
SN - 1433-7851
VL - 62
JO - Angewandte Chemie - International Edition
JF - Angewandte Chemie - International Edition
IS - 9
M1 - e202217372
ER -