Biocatalytic Synthesis of Allylic and Allenyl Sulfides through a Myoglobin-Catalyzed Doyle–Kirmse Reaction

Article abstract of DOI:10.1002/anie.201607278

The first example of a biocatalytic [2,3]-sigmatropic rearrangement reaction involving allylic sulfides and diazo reagents (Doyle–Kirmse reaction) is reported. Engineered variants of sperm whale myoglobin catalyze this synthetically valuable C?C bond-forming transformation with high efficiency and product conversions across a variety of sulfide substrates (e.g., aryl-, benzyl-, and alkyl-substituted allylic sulfides) and α-diazo esters. Moreover, the scope of this myoglobin-mediated transformation could be extended to the conversion of propargylic sulfides to give substituted allenes. Active-site mutations proved effective in enhancing the catalytic efficiency of the hemoprotein in these reactions as well as modulating the enantioselectivity, resulting in the identification of the myoglobin variant Mb(L29S,H64V,V68F), which is capable of mediating asymmetric Doyle–Kirmse reactions with an enantiomeric excess up to 71 %. This work extends the toolbox of currently available biocatalytic strategies for the asymmetric formation of carbon–carbon bonds.

Full text of DOI:10.1002/anie.201607278

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International Edition: DOI: 10.1002/anie.201607278
German Edition: DOI: 10.1002/ange.201607278
Biocatalysis
Biocatalytic Synthesis of Allylic and Allenyl Sulfides through
a Myoglobin-Catalyzed Doyle–Kirmse Reaction
Vikas Tyagi, Gopeekrishnan Sreenilayam, Priyanka Bajaj, Antonio Tinoco, and Rudi Fasan*
Dedicated to Prof. Frances H. Arnold on the occasion of her 60th birthday.
Abstract: The first example of a biocatalytic [2,3]-sigmatropic ester reagents in carbene-transfer reactions. In particular, we
rearrangement reaction involving allylic sulfides and diazo found that engineered variants of sperm whale myoglobin can
reagents (Doyle–Kirmse reaction) is reported. Engineered provide highly active and selective biocatalysts for carbene-
variants of sperm whale myoglobin catalyze this synthetically mediated transformations such as olefin cyclopropanation,[3d]
[9]
À
À
valuable C C bond-forming transformation with high effi- Y H carbene insertion (Y= N, S), and aldehyde olefina-
ciency and product conversions across a variety of sulfide tion.[7] The transition-metal-catalyzed reaction between allyl
substrates (e.g., aryl-, benzyl-, and alkyl-substituted allylic sulfides and a diazo reagent (the Doyle–Kirmse reaction)[10]
À
sulfides) and a-diazo esters. Moreover, the scope of this represents a powerful method for the creation of new C C
myoglobin-mediated transformation could be extended to the bonds, which has found application in the synthesis of various
conversion of propargylic sulfides to give substituted allenes. biologically active molecules.[11] This process involves a reac-
Active-site mutations proved effective in enhancing the cata- tion between the allyl sulfide and a metallo-carbenoid species,
lytic efficiency of the hemoprotein in these reactions as well as leading to the formation of a sulfur ylide, which undergoes
modulating the enantioselectivity, resulting in the identification a [2,3]-sigmatropic rearrangement.[10c,11] While several organ-
of the myoglobin variant Mb(L29S,H64V,V68F), which is ometallic catalysts, including rhodium,[10b,12] copper,[12a–c,13]
capable of mediating asymmetric Doyle–Kirmse reactions with and cobalt[14] complexes, have proven useful for promoting
an enantiomeric excess up to 71%. This work extends the this transformation,[15] the development of catalytically effi-
toolbox of currently available biocatalytic strategies for the cient and enantioselective variants of this reaction has proven
asymmetric formation of carbon–carbon bonds.
very challenging.[11b] Herein, we report the development of
myoglobin-based biocatalysts capable of promoting asym-
B
iocatalytic transformations can provide key opportunities metric Doyle–Kirmse reactions with high catalytic efficiency
for the development of economical and sustainable processes across a broad panel of allylic and propargylic sulfide
for the synthesis and manufacturing of fine chemicals and substrates and different a-diazoesters.
pharmaceuticals.[1] Enzyme-catalyzed carbon–carbon bond-
Initially, we investigated the activity of sperm whale
forming reactions have traditionally involved the use of myoglobin (Mb) in catalyzing the [2,3]-sigmatropic rear-
aldolases, thiamine diphosphate-dependent enzymes, rangement of allyl phenyl sulfide 1 in the presence of ethyl a-
hydroxynitrile lyases, and terpene cyclases, with protein
engineering providing a means to expand the scope of these
enzymes to non-native substrates.[2] More recently, engi-
Table 1: Myoglobin-catalyzed tandem sulfur ylide formation and [2,3]-
sigmatropic rearrangement of phenyl allyl sulfide with EDA.[a]
neered and artificial metalloenzymes have made possible
À
other valuable C C bond-forming transformations, including
olefin cyclopropanation,[3] Suzuki coupling,[4] Diels–Alder
reactions,[5] Friedel–Crafts indole alkylation,[6] Wittig olefina-
tion,[7] and olefin metathesis.[8] Despite this progress, the
toolbox of biocatalytic systems useful for the construction of
À
Entry Catalyst
Deviation from s.r.c. % conv.[b] TON[c]
C C bonds remains limited when compared to synthetic
methods.
1
2
3
4
5
6
7
8
9
WT Mb
WT Mb
WT Mb
WT Mb
–
44%
0%
19%
19%
11%
445
0
195
185
115
>995
3500
6270
>995
Our laboratory and the Arnold group have recently
reported the ability of heme-containing proteins such as
myoglobin[3d,7,9] and P450,[3a–c] respectively, to engage a-diazo
aerobic
no Na2S2O4
1 equiv 2a
0.5 equiv 2a
–
WT Mb
Mb(L29S,H64V,V68F)
Mb(L29S,H64V,V68F) 0.025 mol%
Mb(L29S,H64V,V68F) 0.01 mol%
Mb(L29S,H64V,V68F) 30 min
>99%
87%
[*] Dr. V. Tyagi, Dr. G. Sreenilayam, Dr. P. Bajaj, A. Tinoco,
Prof. Dr. R. Fasan
63%
>99%
Department of Chemistry, University of Rochester
120 Trustee Road, Rochester, NY 14627 (USA)
E-mail: rfasan@ur.rochester.edu
[a] Standard reaction conditions (s.r.c.)=10 mm 1, 20 mm 2a, 10 mm Mb
catalyst (0.1 mol%), 10 mm Na2S2O4 in oxygen-free potassium phos-
phate buffer (50 mm, pH 8) at room temperature. [b] As determined by
gas chromatography. [c] TON=number of turnovers (nmol product/
nmol catalyst). Errors are within Æ10%.
Supporting information and the ORCID identification number(s) for
the author(s) of this article can be found under http://dx.doi.org/10.
1002/anie.201607278.
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diazo acetate (EDA, 2a). Gratifyingly, this reaction resulted
in the formation of the desired [2,3]-sigmatropic rearrange-
ment product 3 with 44% conversion (445 TON) under
optimized conditions (Table 1, Entry 1). No product forma-
tion was observed under aerobic conditions (Table 1,
Entry 2), thus indicating that oxygen, that is, the native
ligand of Mb, inhibits this reactivity. Reactions performed in
the presence and absence of reductant (Na2S2O4) showed that
ferrous Mb is catalytically more efficient than the ferric
counterpart (445 vs. 195 TON, Table 1), although the latter
remains a viable catalyst for this reaction, as supported by
these results and additional experiments (Table S1 and Fig-
ure S2 in the Supporting Information). Varying the pH
between 6 and 9 had a negligible effect on Mb-dependent
catalytic activity, whereas improved conversion was obtained
with a two-fold excess of EDA (55% vs. 19% with one equiv
EDA). As observed for hemin, the Mb-catalyzed formation of
3 show no enantiomeric excess (< 1% ee), thus indicating that
the native Mb scaffold is unable to exert any asymmetric
induction during the reaction.
carbenoid species to yield a chiral sulfur ylide,[12a,13a,c,14b] the
stereochemical information of which is then readily trans-
ferred to the carbon atom during the bond-rearrangement
process.[18] While the native Mb scaffold produces 3 in racemic
form, moderate to good levels of enantioselectivity were
obtained with some of the engineered Mb variants (Fig-
ure S3). Importantly, the highly active Mb(L29S,H64V,V68F)
variant also showed the highest degree of stereocontrol,
yielding 3 with an enantiomeric excess (ee) of 71% (Fig-
ure S3). Notably, Mb(H64V,V68A) favors formation of the
opposite enantiomeric product with 46% ee (49% conv., 490
TON).
To examine the substrate scope of Mb(L29S,H64V,V68F),
variously substituted a-diazo esters and allyl sulfides were
tested. As shown in Table 2, quantitative or nearly quantita-
tive conversions to the desired products 13–15 (94–99%)
were achieved when starting from 1 and diazo reagents such
as tert-butyl (2b), cyclohexyl (2c), or benzyl (2d) a-diazo-
acetate. Good to excellent conversions (57–99%) were also
obtained for reactions involving allyl phenyl sulfides with
substituted phenyl rings (4–6) to give products 16–18. Next,
the Mb(L29S,H64V,V68F)-catalyzed transformation of
benzyl- (8–9) and alkyl-substituted allyl sulfides (10–12) in
the presence of EDA was examined. The high yields
measured for 20 and 21 (78–93%) indicate that benzyl-
substituted allyl sulfides are also efficiently processed by the
biocatalyst. Except for the poorly water-soluble octyl allyl
sulfide (11), moderate to high product conversions (35–86%)
were achieved for the reactions with other alkyl-substituted
allyl sulfides (22, 24), which further supports the broad
substrate scope of Mb(L29S,H64V,V68F). Finally, the suc-
cessful synthesis of 19 from phenyl but-2-enyl sulfide (7) and
EDA (> 99% conv.) showed that substitutions at the level of
the allyl group are also tolerated by the Mb variant. Under
catalyst-limited conditions (i.e., using 0.01 mol%),
Mb(L29S,H64V,V68F) was found to support thousands of
catalytic turnovers (1000–8800) for all of the tested substrates
except 11 (Table 2).
In order to identify more-efficient and selective Mb-based
biocatalysts for this reaction, we evaluated a panel of
engineered Mb variants containing one to three amino acid
substitutions at the level of the five residues defining the distal
cavity of the hemoprotein (Leu29, Phe43, His64, Val68,
Ile107; Figure S1). Previously, we found that mutations at
these positions can dramatically alter the activity and
selectivity of Mb variants as carbene-[3d,7,9,16] and nitrene-[17]
transfer catalysts. Upon testing in the reaction with 1 and
EDA, a number of Mb variants with significantly improved
catalytic activity compared to the wild-type protein were
identified (Figure S3). Among them, Mb(L29S,H64V,V68F)
emerged as the most active biocatalyst for this reaction, giving
quantitative conversion of 1 into 3 at a catalyst loading of only
0.1 mol%. Notably, product conversion values of 87%
(3500 TON) and 63% (6270 TON) were obtained with even
lower catalyst loadings of 0.025 and 0.01 mol%, respectively
(Table 1, Entries 7,8). These results are notable considering
that similar yields in related Doyle–Kirmse reactions have
been achieved with catalyst loadings of 1–5 mol% for Rh-
based complexes[12a,d,f] and 5–20 mol% for synthetic catalysts
based on non-precious metals.[12c,13b,c,14] In addition, in con-
trast to the need for slow addition of the diazo reagent in
The [2,3]-sigmatropic rearrangement of propargylic sul-
fides offers a convenient route to generate allenes, which are
valuable intermediates for a host of synthetic transforma-
tions.[19] To assess the scope of the Mb(L29S,H64V,V68F)
Rh-[12a,d]
and
Cu-catalyzed[13a]
reactions,
the
Mb(L29S,H64V,V68F)-catalyzed reaction proceeds with
excellent chemoselectivity, that is, without carbene dimeriza-
tion, even upon direct mixing of the sulfide and diazo
reactants. Time-course experiments further showed that the
biocatalytic formation of 3 occurs with an initial rate of 167
turnovers per minute (Table S2) and reaches completion
within 30 min (Table 1, Entry 9). These kinetics also compare
very favorably with those of organometallic catalysts, and in
particular those involving Cu and Co complexes, for which
reaction times of 10–36 hours have been reported.[13b,c,14]
Controlling the enantioselectivity of Doyle–Kirmse reac-
tions has proven challenging (typically, < 10–50% ee),
a phenomenon that has been attributed to the difficulty of
discriminating, using chiral catalysts, between the heterotopic
lone pairs of the sulfide during attack on the metallo-
Scheme 1. Mb(L29S,H64V,V68F)-catalyzed [2,3]-sigmatropic rearrange-
ment of propargylic sulfides.
2
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Table 2: Substrate scope of Mb(L29S,H64V,V68F).[a]
Chiral
GC/SFC
analyses
showed
that
Mb(L29S,H64V,V68F) exhibits moderate to good enantiose-
lectivity (20–60% ee) in the transformation of several of the
allylic sulfide substrates of Table 2. In comparison, biocata-
lytic transformation of the propargylic sulfides occurred with
significantly reduced enantiocontrol (< 15% ee; Scheme 1).
These experiments also revealed that the degree of asym-
metric induction can be influenced by the structure of the
diazo reagent (e.g., 47–71% ee for 3 and 15 vs. 6–9% ee for 13
and 14). Finally, a larger-scale reaction with 15 mg of phenyl
allyl sulfide (1), 2 equiv of EDA (2a), and 0.1 mol%
Mb(L29S,H64V,V68F) enabled the isolation of 19.4 mg of 3
in 84% yield, thus demonstrating the scalability of the
biocatalytic process.
Sulfide Diazo reag. Product
% conv. TTN[b] ee [%][c]
(TON)
>99%
(>1000)
1
2b
8170 6%
>99%
(>1000)
1
1
2c
2d
8820 9%
Scheme 2 depicts a plausible mechanism for the Mb-
catalyzed reaction reported herein. We envisage the initial
94%
(940)
3570 47%
>99%
(>1000)
20%
5050
4
5
6
2a
2a
2a
À60%[d]
>99%
(>1000)
5960 40%
57%
(570)
18%
1000
58%[d]
>99%
(>1000)
57/59%
8120
7
8
2a
2a
1:1 d.r.
78%
(780)
7040 10%
Scheme 2. Proposed mechanism and catalytic steps for the myoglobin-
catalyzed Doyle–Kirmse reaction.
93%
(930)
9
2a
5470 43%
3570 38%
formation of an iron-porphyrin-bound carbenoid species (II),
which has electrophilic character[3d,20] and can react with
nucleophiles.[7,9] Accordingly, nucleophilic attack through the
action of the allylic sulfide on this intermediate is envisioned
to give rise to a sulfonium ylide (III), followed by a rapid [2,3]-
sigmatropic rearrangement to yield the final product. The
proposed role of the sulfide substrate as a nucleophile is
consistent with the experimentally observed higher reactivity
of electron-rich allyl sulfides versus isosteric, electrodeficient
counterparts (17 vs. 18; Table 2). This trend was reproduced
with other Mb variants [e.g., 83–95% conversion for 17 vs. 13–
30% for 18 with Mb(H64V,V68A) and Mb(F43V,V68F)],
thus suggesting that the differential reactivity is largely driven
by the electronic properties of the substrate rather than by the
biocatalyst. The enantioselectivity-determining step in asym-
metric Doyle–Kirmse reactions is generally assumed to be
associated with formation of the (chiral) sulfonium yl-
ide,[12a,13a,c,14b] an assumption based on the high degree of
stereoretention observed during the rearrangement of in situ
prepared optically active sulfonium ylides.[18,21] Within this
mechanistic framework, we envision two alternative, although
not mutually exclusive, scenarios by which enantiocontrol
could be exerted by the engineered Mb catalysts: 1) by
35%
(350)
10
11
12
2a
2a
2a
8%
(80)
125
n.d.[e]
86%
(860)
4190 19%
[a] Under standard reactions conditions as described in Table 1.
[b] TTN=total turnover number. Measured using 1 mm Mb catalyst
instead of 10 mm. [c] As determined by chiral GC or Supercritical Fluid
Chromatography (SFC). [d] Using Mb(H64V,V68A). [e] Enantiomers
could not be resolved.
catalyst in the context of this reaction, variously substituted
phenyl propargylic sulfides in combination with ethyl or
benzyl a-diazo-acetate as carbene precursors were tested
(Scheme 1). Notably, the corresponding allenyl-substituted
sulfide products (28–31) were obtained in high yields (71–
83%) in most cases, thereby demonstrating the functionality
of the Mb-based catalyst in promoting the [2,3]-sigmatropic
rearrangement of propargylic sulfide substrates.
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influencing the pre-attack orientation of the sulfide so that
approach to the heme–carbene intermediate through one of
the lone pairs on the sulfur atom is preferred, and/or 2) by
dictating which face of the heme-bound carbenoid group is
exposed to attack by the sulfide nucleophile, in analogy to our
proposed stereochemical model for Mb-catalyzed olefin
cyclopropanation.[3d]
[1] a) U. T. Bornscheuer, G. W. Huisman, R. J. Kazlauskas, S. Lutz,
J. C. Moore, K. Robins, Nature 2012, 485, 185 – 194; b) N. J.
Turner, Nat. Chem. Biol. 2009, 5, 568 – 574; c) A. S. Bommarius,
J. K. Blum, M. J. Abrahamson, Curr. Opin. Chem. Biol. 2011, 15,
194 – 200; d) M. T. Reetz, J. Am. Chem. Soc. 2013, 135, 12480 –
12496.
[2] a) N. G. Schmidt, E. Eger, W. Kroutil, ACS Catal. 2016, 6, 4286 –
4311; b) K. Faber, W. D. Fessner, N. J. Turner, Biocatalysis in
Organic Synthesis. Science of Synthesis, Vol. 1 – 3, Georg Thieme,
Stuttgart, 2015; c) K. Fesko, M. Gruber-Khadjawi, Chem-
CatChem 2013, 5, 1248 – 1272; d) S. C. Hammer, P. O. Syren,
M. Seitz, B. M. Nestl, B. Hauer, Curr. Opin. Chem. Biol. 2013, 17,
293 – 300.
[3] a) P. S. Coelho, E. M. Brustad, A. Kannan, F. H. Arnold, Science
2013, 339, 307 – 310; b) P. S. Coelho, Z. J. Wang, M. E. Ener,
S. A. Baril, A. Kannan, F. H. Arnold, E. M. Brustad, Nat. Chem.
Biol. 2013, 9, 485 – 487; c) Z. J. Wang, H. Renata, N. E. Peck,
C. C. Farwell, P. S. Coelho, F. H. Arnold, Angew. Chem. Int. Ed.
2014, 53, 6810 – 6813; Angew. Chem. 2014, 126, 6928 – 6931;
d) M. Bordeaux, V. Tyagi, R. Fasan, Angew. Chem. Int. Ed. 2015,
54, 1744 – 1748; Angew. Chem. 2015, 127, 1764 – 1768; e) P.
Srivastava, H. Yang, K. Ellis-Guardiola, J. C. Lewis, Nat.
Commun. 2015, 6, 7789; f) J. G. Gober, A. E. Rydeen, E. J.
Gibson-OꢁGrady, J. B. Leuthaeuser, J. S. Fetrow, E. M. Brustad,
ChemBioChem 2016, 17, 394 – 397.
[4] a) S. Abe, J. Niemeyer, M. Abe, Y. Takezawa, T. Ueno, T.
Hikage, G. Erker, Y. Watanabe, J. Am. Chem. Soc. 2008, 130,
10512 – 10514; b) A. Chatterjee, H. Mallin, J. Klehr, J. Vallapur-
ackal, A. D. Finke, L. Vera, M. Marsh, T. R. Ward, Chem. Sci.
2016, 7, 673 – 677.
[5] a) D. Coquiꢂre, J. Bos, J. Beld, G. Roelfes, Angew. Chem. Int. Ed.
2009, 48, 5159 – 5162; Angew. Chem. 2009, 121, 5261 – 5264; b) J.
Podtetenieff, A. Taglieber, E. Bill, E. J. Reijerse, M. T. Reetz,
Angew. Chem. Int. Ed. 2010, 49, 5151 – 5155; Angew. Chem.
2010, 122, 5277 – 5281.
[6] J. Bos, W. R. Browne, A. J. Driessen, G. Roelfes, J. Am. Chem.
Soc. 2015, 137, 9796 – 9799.
[7] V. Tyagi, R. Fasan, Angew. Chem. Int. Ed. 2016, 55, 2512 – 2516;
Angew. Chem. 2016, 128, 2558 – 2562.
[8] a) C. Mayer, D. G. Gillingham, T. R. Ward, D. Hilvert, Chem.
Commun. 2011, 47, 12068 – 12070; b) F. Philippart, M. Arlt, S.
Gotzen, S. J. Tenne, M. Bocola, H. H. Chen, L. Zhu, U.
Schwaneberg, J. Okuda, Chem. Eur. J. 2013, 19, 13865 – 13871;
c) M. Basauri-Molina, D. G. A. Verhoeven, A. J. van Schaik, H.
Kleijn, R. J. M. K. Gebbink, Chem. Eur. J. 2015, 21, 15676 –
15685.
[9] a) G. Sreenilayam, R. Fasan, Chem. Commun. 2015, 51, 1532 –
1534; b) V. Tyagi, R. B. Bonn, R. Fasan, Chem. Sci. 2015, 6,
2488 – 2494.
While further studies are warranted to discriminate
between these scenarios, experiments were performed to
clarify the beneficial role of the active-site mutations in
Mb(L29S,H64V,V68F). To this end, we characterized and
compared the catalytic activity and selectivity of a set of single
and double reversion mutants in the reaction with phenyl allyl
sulfide (1) and EDA (Table S2). In line with our previous
observations,[3d,9a,16] mutation of the distal His residue (H64V)
increases both the TTN (2230 vs. 1615 for Mb) and product
formation rate (121 vs. 79 minÀ1), possibly through facilitating
access of the substrate to the heme cavity (Figure S1). The
L29S mutation further enhances the catalytic competency of
the hemoprotein, as suggested by the increased total turn-
overs (3085 TTN) supported by Mb(L29S,H64V). The V68F
mutation has no beneficial effect when used alone or in
combination with H64V, but it contributes synergistically with
L29S
to
improving
both
TTN
[6280
for
Mb(L29S,H64V,V68F) vs. 3085 for Mb(L29S,H64V)] and
rate (167 vs. 118 minÀ1, respectively). Interestingly, none of
the single-site or double-site variants showed significantly
improved enantioselectivity compared to wild-type Mb
(< 5% ee; Table S2). These results indicate that the enhanced
enantioselectivity of Mb(L29S,H64V,V68F) stems from a syn-
ergistic contribution from the three active-site mutations. This
effect is likely facilitated by the close proximity of these
residues within the heme pocket (ca. 7 ꢀ for C(b)···C(b)
distance; Figure S1).
In summary, this study demonstrates that engineered Mb
variants can serve as efficient biocatalysts for asymmetric
Doyle–Kirmse reactions. When using the optimized variant
Mb(L29S,H64V,V68F), good to excellent product conver-
sions as well as high numbers of catalytic turnovers (up to
8820) were achieved across a variety of allylic and propargylic
sulfides in the presence of a-diazo ester-based carbene
precursors. Importantly, the enantioselectivity of the Mb
catalyst could be tuned and optimized through mutations
within the distal pocket of the protein. This work expands the
toolbox of biocatalytic strategies for mediating sigmatropic
rearrangements[22] and the asymmetric formation of carbon–
carbon bonds.[2a]
[10] a) W. Kirmse, M. Kapps, Chem. Ber. 1968, 101, 994 – 996;
b) M. P. Doyle, J. H. Griffin, M. S. Chinn, D. Vanleusen, J. Org.
Chem. 1984, 49, 1917 – 1925; c) M. P. Doyle, D. C. Forbes, Chem.
Rev. 1998, 98, 911 – 936.
Acknowledgements
This work was supported by the U.S. National Institute of
Health grant GM098628. We are grateful to Prof. Daniel Weix
(U. Rochester) for providing access to the SFC instrumenta-
tion. LC-MS instrumentation was supported by the U.S. NSF
grant CHE-0946653.
[11] a) J. B. Sweeney, Chem. Soc. Rev. 2009, 38, 1027 – 1038; b) T. H.
West, S. S. M. Spoehrle, K. Kasten, J. E. Taylor, A. D. Smith,
ACS Catal. 2015, 5, 7446 – 7479.
[12] a) Y. Nishibayashi, K. Ohe, S. Uemura, J. Chem. Soc. Chem.
Commun. 1995, 1245 – 1246; b) V. K. Aggarwal, M. Ferrara, R.
Hainz, S. E. Spey, Tetrahedron Lett. 1999, 40, 8923 – 8927;
c) X. M. Zhang, M. Ma, J. B. Wang, Tetrahedron: Asymmetry
2003, 14, 891 – 895; d) A. G. H. Wee, Q. Shi, Z. Y. Wang, K.
Hatton, Tetrahedron: Asymmetry 2003, 14, 897 – 909; e) P.
Keywords: biocatalysis · heme proteins · protein engineering ·
sigmatropic rearrangement · ylides
4
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Mꢃller, S. Grass, S. P. Shahi, G. Bernardinelli, Tetrahedron 2004,
[20] a) R. L. Khade, W. C. Fan, Y. Ling, L. Yang, E. Oldfield, Y.
60, 4755 – 4763; f) M. Liao, B. Wang, Green Chem. 2007, 9, 184 –
188; g) H. Zhang, B. Wang, H. Yi, Y. Zhang, J. B. Wang, Org.
Lett. 2015, 17, 3322 – 3325.
Zhang, Angew. Chem. Int. Ed. 2014, 53, 7574 – 7578; Angew.
Chem. 2014, 126, 7704 – 7708; b) J. R. Wolf, C. G. Hamaker, J. P.
Djukic, T. Kodadek, L. K. Woo, J. Am. Chem. Soc. 1995, 117,
9194 – 9199.
[13] a) D. W. McMillen, N. Varga, B. A. Reed, C. King, J. Org. Chem.
2000, 65, 2532 – 2536; b) X. M. Zhang, Z. H. Qu, Z. H. Ma, W. F.
Shi, X. L. Jin, J. B. Wang, J. Org. Chem. 2002, 67, 5621 – 5625;
c) M. Ma, L. L. Peng, C. K. Li, X. Zhang, J. B. Wang, J. Am.
Chem. Soc. 2005, 127, 15016 – 15017.
[14] a) T. Fukuda, T. Katsuki, Tetrahedron Lett. 1997, 38, 3435 – 3438;
b) T. Fukuda, R. Irie, T. Katsuki, Tetrahedron 1999, 55, 649 – 664.
[15] a) C. Y. Zhou, W. Y. Yu, P. W. H. Chan, C. M. Che, J. Org. Chem.
2004, 69, 7072 – 7082; b) P. W. Davies, S. J. C. Albrecht, G.
Assanelli, Org. Biomol. Chem. 2009, 7, 1276 – 1279; c) D. S.
Carter, D. L. Van Vranken, Org. Lett. 2000, 2, 1303 – 1305;
d) M. S. Holzwarth, I. Alt, B. Plietker, Angew. Chem. Int. Ed.
2012, 51, 5351 – 5354; Angew. Chem. 2012, 124, 5447 – 5450.
[16] S. Giovani, R. Singh, R. Fasan, Chem. Sci. 2016, 7, 234 – 239.
[17] M. Bordeaux, R. Singh, R. Fasan, Bioorg. Med. Chem. 2014, 22,
5697 – 5704.
[21] B. M. Trost, W. G. Biddlecom, J. Org. Chem. 1973, 38, 3438 –
3439.
[22] a) D. Hilvert, S. H. Carpenter, K. D. Nared, M. T. M. Auditor,
Proc. Natl. Acad. Sci. USA 1988, 85, 4953 – 4955; b) D. Y.
Jackson, J. W. Jacobs, R. Sugasawara, S. H. Reich, P. A. Bartlett,
P. G. Schultz, J. Am. Chem. Soc. 1988, 110, 4841 – 4842; c) M. R.
Haynes, E. A. Stura, D. Hilvert, I. A. Wilson, Science 1994, 263,
646 – 652; d) S. S. Yoon, Y. Oei, E. Sweet, P. G. Schultz, J. Am.
Chem. Soc. 1996, 118, 11686 – 11687; e) H. D. Ulrich, E. Mundr-
off, B. D. Santarsiero, E. M. Driggers, R. C. Stevens, P. G.
Schultz, Nature 1997, 389, 271 – 275; f) Z. H. S. Zhou, A. Flohr,
D. Hilvert, J. Org. Chem. 1999, 64, 8334 – 8341; g) C. K. Prier,
T. K. Hyster, C. C. Farwell, A. Huang, F. H. Arnold, Angew.
Chem. Int. Ed. 2016, 55, 4711 – 4715; Angew. Chem. 2016, 128,
4789 – 4793.
[18] B. M. Trost, R. F. Hammen, J. Am. Chem. Soc. 1973, 95, 962 –
964.
[19] Modern Allene Chemistry (Eds.: N. Krause, A. S. K. Hashmi),
Wiley-VCH, Weinheim, 2004.
Received: July 27, 2016
Published online: && &&, &&&&
Angew. Chem. Int. Ed. 2016, 55, 1 – 6
ꢀ 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
www.angewandte.org
5
These are not the final page numbers!

Angewandte
Communications
Chemie
Communications
Biocatalysis
V. Tyagi, G. Sreenilayam, P. Bajaj,
A. Tinoco, R. Fasan*
&&&& — &&&&
Biocatalytic Synthesis of Allylic and
Allenyl Sulfides through a Myoglobin-
Catalyzed Doyle–Kirmse Reaction
Biocatalytic Doyle-Kirmse reaction: Engi-
neered variants of sperm whale myoglo-
bin catalyze the tandem sulfur ylide
formation/[2,3]-sigmatropic rearrange-
ment of allylic and propargylic sulfides in
the presence of a-diazo esters. This
transformation can be applied to a variety
of aryl, benzylic, and alkyl sulfide sub-
strates, and gives high product conver-
sion and enantioselectivity up to 71% ee.
6
www.angewandte.org
ꢀ 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Angew. Chem. Int. Ed. 2016, 55, 1 – 6
These are not the final page numbers!