TY - JOUR

T1 - Convergent Cascade Catalyzed by Monooxygenase–Alcohol Dehydrogenase Fusion Applied in Organic Media

AU - Huang, Lei

AU - Aalbers, Friso S.

AU - Tang, Wei

AU - Röllig, Robert

AU - Fraaije, Marco W.

AU - Kara, Selin

N1 - Funding Information: S.K. thanks the Fonds der Chemischen Industrie (Frankfurt, Germany) for financial support (grant no. SK 196/20). L.H. thanks the China Scholarship Council (CSC) for financial support. F.S.A. and M.W.F. received funding from the European Union (EU) project ROBOX (grant agreement no. 635734), the EU's Horizon 2020 Program Research and Innovation actions H2020-LEIT BIO-2014-1. The views and opinions expressed in this article are only those of the authors and do not necessarily reflect those of the European Union Research Agency. The European Union is not liable for any use that may be made of the information contained herein. We thank Dr. Diederik Johannes Opperman (University of the Free State, South Africa) for the recombinant plasmid containing HLADH gene. R.R. contributed to some parts of the experimental work and interpretation of the data as an intern student from the research group of Prof. Dr. Marion Ansorge-Schumacher (Chair of Molecular Biotechnology, TU Dresden). Funding Information: S.K. thanks the Fonds der Chemischen Industrie (Frankfurt, Germany) for financial support (grant no. SK 196/20). L.H. thanks the China Scholarship Council (CSC) for financial support. F.S.A. and M.W.F. received funding from the European Union (EU) project ROBOX (grant agreement no. 635734), the EU’s Horizon 2020 Program Research and Innovation actions H2020-LEIT BIO-2014-1.

PY - 2019/6/30

Y1 - 2019/6/30

N2 - With the aim of applying redox-neutral cascade reactions in organic media, fusions of a type II flavin-containing monooxygenase (FMO-E) and horse liver alcohol dehydrogenase (HLADH) were designed. The enzyme orientation and expression vector were found to influence the overall fusion enzyme activity. The resulting bifunctional enzyme retained the catalytic properties of both individual enzymes. The lyophilized cell-free extract containing the bifunctional enzyme was applied for the convergent cascade reaction consisting of cyclobutanone and butane-1,4-diol in different microaqueous media with only 5 % (v/v) aqueous buffer without any addition of external cofactor. Methyl tert-butyl ether and cyclopentyl methyl ether were found to be the best organic media for the synthesis of γ-butyrolactone, resulting in about 27 % analytical yield.

AB - With the aim of applying redox-neutral cascade reactions in organic media, fusions of a type II flavin-containing monooxygenase (FMO-E) and horse liver alcohol dehydrogenase (HLADH) were designed. The enzyme orientation and expression vector were found to influence the overall fusion enzyme activity. The resulting bifunctional enzyme retained the catalytic properties of both individual enzymes. The lyophilized cell-free extract containing the bifunctional enzyme was applied for the convergent cascade reaction consisting of cyclobutanone and butane-1,4-diol in different microaqueous media with only 5 % (v/v) aqueous buffer without any addition of external cofactor. Methyl tert-butyl ether and cyclopentyl methyl ether were found to be the best organic media for the synthesis of γ-butyrolactone, resulting in about 27 % analytical yield.

KW - biocatalysis

KW - domino reactions

KW - enzymes

KW - fusion enzymes

KW - solvent effects

UR - http://www.scopus.com/inward/record.url?scp=85066911767&partnerID=8YFLogxK

U2 - 10.1002/cbic.201800814

DO - 10.1002/cbic.201800814

M3 - Article

C2 - 30811825

AN - SCOPUS:85066911767

VL - 20

SP - 1653

EP - 1658

JO - CHEMBIOCHEM

JF - CHEMBIOCHEM

SN - 1439-4227

IS - 13

ER -