Please do not adjust margins
Organic & Biomolecular Chemistry
Page 4 of 5
DOI: 10.1039/C7OB00390K
ARTICLE
Journal Name
‡‡Interestingly, this mutant also showed some background activity
in absence of Cu2+, albeit that the catalysis was significantly
improved in the presence of Cu(NO3)2 (see Table S3) The origin of
this background activity is not understood at present.
Table 2 Scope of vinylogous Friedel−Craꢀs alkylaꢁon reacꢁons catalyzed by QacR
Y123BpyA_Cu2+
1
2
3
J. C. Lewis, ACS Catal., 2013, 3, 2954–2975.
J. Bos and G. Roelfes, Curr. Opin. Chem. Biol., 2014, 19, 135–143.
O. Pàmies, M. Diéguez and J.-E. Bäckvall, Adv. Synth. Catal.,
2015, 357, 1567–1586.
4
5
6
E. M. Brustad and F. H. Arnold, Curr. Opin. Chem. Biol., 2011, 15,
201–210.
H. Agustiandari, J. Lubelski, H. B. van den B. van Saparoea, O. P.
Kuipers and A. J. M. Driessen, J. Bacteriol., 2008, 190, 759–763.
P. K. Madoori, H. Agustiandari, A. J. M. Driessen and A.-M. W. H.
Thunnissen, EMBO J., 2009, 28, 156–166.
H. Wade, Curr. Opin. Struct. Biol., 2010, 20, 489–496.
J. Bos, F. Fusetti, A. J. M. Driessen and G. Roelfes, Angew. Chem.
Int. Ed., 2012, 51, 7472–7475.
J. Bos, A. García-Herraiz and G. Roelfes, Chem. Sci., 2013, 4,
3578–3582.
Yield
Entry Indole
1
R
Catalyst
Cu(NO3)2
Product
ee (%)c
<5
14±3
33±13
14±3
2
QacR_Cu2+
27±3 (-)
<5
R1=H
R2=H
2a
3a
3
QacR_Y123BpyA_Cu2+
7
8
4
Cu(NO3)2
QacR_Cu2+
6±2
20±9
3±0
<5
R1=Me
R2=H
5
6
2b
2c
2d
3b
3c
3d
58±2 (-)
56±6 (+)
9
QacR_Y123BpyA_Cu2+
10 I. Drienovská, A. Rioz-Martínez, A. Draksharapu and G. Roelfes,
Chem. Sci., 2014, 6, 770–776.
11 J. Bos, W. R. Browne, A. J. M. Driessen and G. Roelfes, J. Am.
Chem. Soc., 2015, 137, 9796–9799.
12 L. Wang, A. Brock, B. Herberich and P. G. Schultz, Science, 2001,
292, 498–500.
13 J. Xie, W. Liu and P. G. Schultz, Angew. Chem. Int. Ed., 2007, 46,
9239–9242.
7
8
9
Cu(NO3)2
QacR_Cu2+
26±1
58±17
26±3
<5
<5
R1=H
R2=OMe
QacR_Y123BpyA_Cu2+
12±5 (+)
10
11
12
Cu(NO3)2
QacR_Cu2+
3±1
21±7
5±2
<5
R1=H
R2=Cl
47±7 (-)
33±3 (+)
14 H. S. Lee, G. Spraggon, P. G. Schultz and F. Wang, J. Am. Chem.
Soc., 2009, 131, 2481–2483.
QacR_Y123BpyA_Cu2+
Same reaction conditions as in Table 1 a Yields were determined by HPLC and using
2-phenylquinoline as internal standard. Errors listed are standard deviations. b Sign
of rotation was assigned based on the elution order in chiral HPLC by comparison
to previous reports 34,36
15 S. Grkovic, M. H. Brown, N. J. Roberts, I. T. Paulsen and R. A.
Skurray, J. Biol. Chem., 1998, 273, 18665–18673.
16 M. A. Schumacher, M. C. Miller, S. Grkovic, M. H. Brown, R. A.
Skurray and R. G. Brennan, Science, 2001, 294, 2158–2163.
17 S. Grkovic, K. M. Hardie, M. H. Brown and R. A. Skurray,
Biochemistry (Mosc.), 2003, 42, 15226–15236.
18 K. M. Peters, G. Sharbeen, T. Theis, R. A. Skurray and M. H.
Brown, Biochemistry (Mosc.), 2009, 48, 9794–9800.
19 H. Itou, U. Okada, H. Suzuki, M. Yao, M. Wachi, N. Watanabe
and I. Tanaka, J. Biol. Chem., 2005, 280, 38711–38719.
20 S. Yamasaki, E. Nikaido, R. Nakashima, K. Sakurai, D. Fujiwara, I.
Fujii and K. Nishino, Nat. Commun., 2013, 4.
Interestingly this mutant afforded the opposite enantiomer
compared to other QacR mutants as well as to previous LmrR-based
metalloenzymes, which is from the PadR family of MDRs.10,11
This work illustrates that MDRs of the TetR family are an attractive
class of scaffolds for artificial metalloenzyme design. Thus, when
combined with our earlier work on LmrR, this shows that MDRs can
be seen as a general platform for the design and construction of
hybrid catalysts that readily available for evaluation in diverse
catalytic reactions.
21 H. Itou, N. Watanabe, M. Yao, Y. Shirakihara and I. Tanaka, J.
Mol. Biol., 2010, 403, 174–184.
22 K. Takeuchi, Y. Tokunaga, M. Imai, H. Takahashi and I. Shimada,
Sci. Rep., 2014, 4.
Acknowledgements
23 N. Park, J. Ryu, S. Jang and H. S. Lee, Tetrahedron, 2012, 68,
4649–4654.
24 M. Kang, K. Light, H. Ai, W. Shen, C. H. Kim, P. R. Chen, H. S. Lee,
E. I. Solomon and P. G. Schultz, ChemBioChem, 2014, 15, 822–
825.
25 W. Ko, S. Kim, K. Jo and H. S. Lee, Amino Acids, 2016, 48, 357–
363.
26 X. Luo, T.-S. A. Wang, Y. Zhang, F. Wang and P. G. Schultz, Cell
Chem. Biol., 2016, 23, 1098–1102.
This work was supported by the European Research Council (ERC
Starting Grant 280010). Financial support from the Ministry of
Education, Culture, and Science (Gravitation Program No.
024.001.035) is gratefully acknowledged. The authors thank Prof. P.
G. Schultz (The Scripps Research Institute) for kindly providing the
pEVOL plasmid for in vivo incorporation of BpyA. The authors wish
to thank Ivana Drienovská and Annika Borg for useful suggestions
and discussion.
27 R. Ballardini, V. Balzani, M. Clemente-León, A. Credi, M. T.
Gandolfi, E. Ishow, J. Perkins, J. F. Stoddart, H.-R. Tseng and S.
Wenger, J. Am. Chem. Soc., 2002, 124, 12786–12795.
28 S. Grkovic, M. H. Brown, M. A. Schumacher, R. G. Brennan and
R. A. Skurray, J. Bacteriol., 2001, 183, 7102–7109.
29 K. M. Peters, B. E. Brooks, M. A. Schumacher, R. A. Skurray, R. G.
Brennan and M. H. Brown, PLOS ONE, 2011, 6, e15974.
Notes and references
‡Fitting of the titration curves for RamR Y92BpyA and CgmR
L100BpyA were not performed due to precipitation of the protein
after addition of more than 1 eq of Cu2+ or difficulties with reliably
of fitting the data, respectively
4 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins