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Production of Propene from n ‐Butanol: A Three‐Step Cascade Utilizing the Cytochrome P450 Fatty Acid Decarboxylase OleT JE
Author(s) -
Bauer Daniel,
Zachos Ioannis,
Sieber Volker
Publication year - 2020
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.202000378
Subject(s) - cofactor , decarboxylation , chemistry , nad+ kinase , alcohol dehydrogenase , fatty acid , oxidative decarboxylation , fatty acid metabolism , butyric acid , biochemistry , cytochrome p450 , stereochemistry , enzyme , catalysis
Propene is one of the most important starting materials in the chemical industry. Herein, we report an enzymatic cascade reaction for the biocatalytic production of propene starting from n ‐butanol, thus offering a biobased production from glucose. In order to create an efficient system, we faced the issue of an optimal cofactor supply for the fatty acid decarboxylase OleT JE , which is said to be driven by either NAD(P)H or H 2 O 2 . In the first system, we used an alcohol and aldehyde dehydrogenase coupled to OleT JE by the electron‐transfer complex putidaredoxin reductase/putidaredoxin, allowing regeneration of the NAD + cofactor. With the second system, we intended full oxidation of n ‐butanol to butyric acid, generating one equivalent of H 2 O 2 that can be used for the oxidative decarboxylation. As the optimal substrate is a long‐chain fatty acid, we also tried to create an improved variant for the decarboxylation of butyric acid by using rational protein design. Within a mutational study with 57 designed mutants, we generated the mutant OleT V292I , which showed a 2.4‐fold improvement in propene production in our H 2 O 2 ‐driven cascade system and reached total turnover numbers >1000.