Structure-Based Insight into the Asymmetric Bioreduction of the C=C Double Bond of alpha,beta-Unsaturated Nitroalkenes by Pentaerythritol Tetranitrate Reductase.
Biocatalytic reduction of alpha- or beta-alkyl-beta-arylnitroalkenes provides a convenient and efficient method to prepare chiral substituted nitroalkanes. Pentaerythritol tetranitrate reductase (PETN reductase) from Enterobacter cloacae st. PB2 catalyses the reduction of nitroolefins such as 1-nitrocyclohexene (1) with steady state and rapid reaction kinetics comparable to other old yellow enzyme homologues. Furthermore, it reduces 2-aryl-1-nitropropenes (4a-d) to their equivalent (S)-nitropropanes 9a-d. The enzyme shows a preference for the (Z)-isomer of substrates 4a-d, providing almost pure enantiomeric products 9a-d (ees up to > 99%) in quantitative yield, whereas the respective (E)-isomers are reduced with lower enantioselectivity (63-89% ee) and lower product yields. 1-Aryl-2-nitropropenes (5a, b) are also reduced efficiently, but the products (R)-10 have lower optical purities. The structure of the enzyme complex with 1-nitrocyclohexene (1) was determined by X-ray crystallography, revealing two substrate-binding modes, with only one compatible with hydride transfer. Models of nitropropenes 4 and 5 in the active site of PETN reductase predicted that the enantioselectivity of the reaction was dependent on the orientation of binding of the (E)- and (Z)-substrates. This work provides a structural basis for understanding the mechanism of asymmetric bioreduction of nitroalkenes by PETN reductase.
Toogood
Helen S
HS
Manchester Interdisciplinary Biocentre, Faculty of Life Sciences, University of Manchester, 131 Princess Street, Manchester M1 7DN, U.K.
Fryszkowska
Anna
A
Hare
Victoria
V
Fisher
Karl
K
Roujeinikova
Anna
A
Leys
David
D
Gardiner
John M
JM
Stephens
Gill M
GM
Scrutton
Nigel S
NS
Acta Crystallogr D Biol Crystallogr. 1998 Jul 1;54(Pt 4):675-7
9761872
J Biol Chem. 2004 Jul 16;279(29):30563-72
15128738
J Biol Chem. 2005 Jul 22;280(29):27103-10
15905167
Org Lett. 2004 Nov 25;6(24):4575-7
15548079
Angew Chem Int Ed Engl. 2007;46(21):3934-7
17431865
J Biol Chem. 1987 Oct 25;262(30):14428-34
3667582
Org Lett. 2007 Dec 20;9(26):5409-11
18031047
Org Lett. 2006 Dec 21;8(26):6131-3
17165947
Structure. 1994 Nov 15;2(11):1089-105
7881908
J Biol Chem. 2002 Jun 14;277(24):21906-12
11923299
Nat Struct Biol. 1999 May;6(5):458-63
10331874
Appl Environ Microbiol. 2004 Jun;70(6):3566-74
15184158
J Biol Chem. 1983 Aug 25;258(16):9913-24
6885775
J Comput Chem. 2005 Dec;26(16):1668-88
16200636
J Comput Chem. 2004 Jul 15;25(9):1157-74
15116359
Chem Rev. 2005 Mar;105(3):933-71
15755081
Angew Chem Int Ed Engl. 2007;46(18):3316-8
17387664
J Biol Chem. 2005 Mar 18;280(11):10695-709
15632179
Proc Natl Acad Sci U S A. 2002 Mar 5;99(5):2702-7
11867731
J Bacteriol. 1996 Nov;178(22):6623-7
8932320
Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240-55
15299926
J Mol Biol. 2001 Jul 6;310(2):433-47
11428899
J Biol Chem. 1975 Jun 10;250(11):4048-52
1126943
J Biol Chem. 2001 Jul 6;276(27):24790-6
11333274
Curr Opin Chem Biol. 2007 Apr;11(2):203-13
17353140
J Biol Chem. 2001 Feb 23;276(8):5779-87
11060310
Biotechnol Bioeng. 2007 Sep 1;98(1):22-9
17657768
J Am Chem Soc. 2005 Apr 6;127(13):4584-5
15796518
FEBS J. 2005 Sep;272(18):4660-71
16156787
J Biol Chem. 1998 Dec 4;273(49):32763-70
9830020
J Org Chem. 2008 Jun 6;73(11):4295-8
18452336
Microbiology (Reading). 2002 Jun;148(Pt 6):1607-1614
12055282
J Am Chem Soc. 2007 Jul 25;129(29):8976-7
17602561
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
15572765
Angew Chem Int Ed Engl. 2003 Oct 13;42(39):4793-5
14562351
Trends Biochem Sci. 2000 Mar;25(3):126-32
10694883
J Am Chem Soc. 2008 Jun 18;130(24):7655-8
18500801
Biochemistry. 1995 Apr 4;34(13):4246-56
7703238
J Biol Chem. 2005 Feb 18;280(7):5195-204
15582992
Org Lett. 2004 Aug 5;6(16):2829-32
15281780
J Biol Chem. 2003 Nov 7;278(45):43973-82
12941965
Proc Natl Acad Sci U S A. 2000 Sep 26;97(20):10733-8
10995477
J Biol Chem. 1964 Nov;239:3927-34
14257628