A designer enzyme for hydrazone and oxime formation featuring an unnatural catalytic aniline residue

被引:124
作者
Drienovska, Ivana [1 ]
Mayer, Clemens [1 ]
Dulson, Christopher [1 ]
Roelfes, Gerard [1 ]
机构
[1] Univ Groningen, Stratingh Inst Chem, Groningen, Netherlands
基金
欧洲研究理事会;
关键词
DIELS-ALDER REACTION; NOVO PROTEIN DESIGN; ARTIFICIAL METALLOENZYMES; NUCLEOPHILIC CATALYSIS; GENETIC-CODE; AMINO-ACIDS; ANTIBODY CATALYSIS; LMRR; CHEMISTRY; MECHANISM;
D O I
10.1038/s41557-018-0082-z
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
Creating designer enzymes with the ability to catalyse abiological transformations is a formidable challenge. Efforts toward this goal typically consider only canonical amino acids in the initial design process. However, incorporating unnatural amino acids that feature uniquely reactive side chains could significantly expand the catalytic repertoire of designer enzymes. To explore the potential of such artificial building blocks for enzyme design, here we selected p-aminophenylalanine as a potentially novel catalytic residue. We demonstrate that the catalytic activity of the aniline side chain for hydrazone and oxime formation reactions is increased by embedding p-aminophenylalanine into the hydrophobic pore of the multidrug transcriptional regulator from Lactococcus lactis. Both the recruitment of reactants by the promiscuous binding pocket and a judiciously placed aniline that functions as a catalytic residue contribute to the success of the identified artificial enzyme. We anticipate that our design strategy will prove rewarding to significantly expand the catalytic repertoire of designer enzymes in the future.
引用
收藏
页码:946 / 952
页数:7
相关论文
共 41 条
[1]   Biocatalysis with Unnatural Amino Acids: Enzymology Meets Xenobiology [J].
Agostini, Federica ;
Voeller, Jan-Stefan ;
Koksch, Beate ;
Acevedo-Rocha, Carlos G. ;
Kubyshkin, Vladimir ;
Budisa, Nediljko .
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2017, 56 (33) :9680-9703
[2]   LmrR is a transcriptional repressor of expression of the multidrug ABC transporter lmrCD in Lactococcus lactis [J].
Agustiandari, Herfita ;
Lubelski, Jacek ;
van Saparoea, H. Bart van den Berg ;
Kuipers, Oscar P. ;
Driessen, Arnold J. M. .
JOURNAL OF BACTERIOLOGY, 2008, 190 (02) :759-763
[3]   Albumin as a promiscuous biocatalyst in organic synthesis [J].
Albanese, Domenico C. M. ;
Gaggero, Nicoletta .
RSC ADVANCES, 2015, 5 (14) :10588-10598
[4]   Formylglycine, a Post-Translationally Generated Residue with Unique Catalytic Capabilities and Biotechnology Applications [J].
Appel, Mason J. ;
Bertozzi, Carolyn R. .
ACS CHEMICAL BIOLOGY, 2015, 10 (01) :72-84
[5]   Multidrug resistance regulators (MDRs) as scaffolds for the design of artificial metalloenzymes [J].
Bersellini, Manuela ;
Roelfes, Gerard .
ORGANIC & BIOMOLECULAR CHEMISTRY, 2017, 15 (14) :3069-3073
[6]   Supramolecular Assembly of Artificial Metalloenzymes Based on the Dimeric Protein LmrR as Promiscuous Scaffold [J].
Bos, Jeffrey ;
Browne, Wesley R. ;
Driessen, Arnold J. M. ;
Roelfes, Gerard .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2015, 137 (31) :9796-9799
[7]   Enantioselective Artificial Metalloenzymes by Creation of a Novel Active Site at the Protein Dimer Interface [J].
Bos, Jeffrey ;
Fusetti, Fabrizia ;
Driessen, Arnold J. M. ;
Roelfes, Gerard .
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2012, 51 (30) :7472-7475
[8]   AN ANTIBODY-CATALYZED BIMOLECULAR DIELS-ALDER REACTION [J].
BRAISTED, AC ;
SCHULTZ, PG .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1990, 112 (20) :7430-7431
[9]   Oxidative coupling of peptides to a virus capsid containing unnatural amino acids [J].
Carrico, Zachary M. ;
Romanini, Dante W. ;
Mehl, Ryan A. ;
Francis, Matthew B. .
CHEMICAL COMMUNICATIONS, 2008, (10) :1205-1207
[10]   Addition of p-azido-L-phenylaianine to the genetic code of Escherichia coli [J].
Chin, JW ;
Santoro, SW ;
Martin, AB ;
King, DS ;
Wang, L ;
Schultz, PG .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2002, 124 (31) :9026-9027