Controllable stereoinversion in DNA-catalyzed olefin cyclopropanationviacofactor modification

Article abstract of DOI:10.1039/d1sc00755f

The assembly of DNA with metal-complex cofactors can form promising biocatalysts for asymmetric reactions, although catalytic performance is typically limited by low enantioselectivities and stereo-control remains a challenge. Here, we engineer G-quadruplex-based DNA biocatalysts for an asymmetric cyclopropanation reaction, achieving enantiomeric excess (ee_trans) values of up to +91% with controllable stereoinversion, where the enantioselectivity switches to ?72% ee_transthrough modification of the Fe-porphyrin cofactor. Complementary circular dichroism, nuclear magnetic resonance, and fluorescence titration experiments show that the porphyrin ligand of the cofactor participates in the regulation of the catalytic enantioselectivityviaa synergetic effect with DNA residues at the active site. These findings underline the important role of cofactor modification in DNA catalysis and thus pave the way for the rational engineering of DNA-based biocatalysts.

Full text of DOI:10.1039/d1sc00755f

Chemical
Science
View Article Online
View Journal | View Issue
EDGE ARTICLE
Controllable stereoinversion in DNA-catalyzed
olefin cyclopropanation via cofactor modification†
Cite this: Chem. Sci., 2021, 12, 7918
Jingya Hao,ab Wenhui Miao,ab Shengmei Lu,a Yu Cheng,ab Guoqing Jiaa
All publication charges for this article
have been paid for by the Royal Society
of Chemistry
a
*
and Can Li
The assembly of DNA with metal-complex cofactors can form promising biocatalysts for asymmetric
reactions, although catalytic performance is typically limited by low enantioselectivities and stereo-
control remains
a challenge. Here, we engineer G-quadruplex-based DNA biocatalysts for an
asymmetric cyclopropanation reaction, achieving enantiomeric excess (eetrans) values of up to +91% with
controllable stereoinversion, where the enantioselectivity switches to ꢀ72% eetrans through modification
of the Fe-porphyrin cofactor. Complementary circular dichroism, nuclear magnetic resonance, and
fluorescence titration experiments show that the porphyrin ligand of the cofactor participates in the
regulation of the catalytic enantioselectivity via a synergetic effect with DNA residues at the active site.
These findings underline the important role of cofactor modification in DNA catalysis and thus pave the
way for the rational engineering of DNA-based biocatalysts.
Received 7th February 2021
Accepted 23rd April 2021
DOI: 10.1039/d1sc00755f
rsc.li/chemical-science
Roelfes32 and Sen33 groups expanded the scope of reactions
catalyzed by DNA-based biocatalysts to include olen cyclo-
Introduction
propanation. Although the enantioselectivities achieved were
moderate, this has paved the way for DNA-catalyzed carbene
transfer reactions. Therefore, designing a DNA-based bio-
catalyst that catalyzes cyclopropanation in high enantiomeric
excess (ee) remains a challenge, especially to achieve an ee
greater than 90%.34
Cyclopropane motifs feature in many natural products and
medicinal agents1 which constitute versatile intermediates for
the total synthesis of therapeutic compounds.2 As these
compounds are in high demand, signicant effort has been
devoted to the development of cyclopropane synthesis, in
particular via the use of hemoprotein enzymes engineered via
directed evolution. Cytochrome P450 (ref. 3 and 4) and
myoglobin5–8 enzymes have been evolved to catalyze asymmetric
olen cyclopropanations with excellent performance.9–14 This
method has been applied to the synthesis of drug molecules15,16
and natural product scaffolds.17,18
Despite this progress, the developed biocatalytic protocols
are generally restricted to protein enzyme engineering.19 The
discovery of the catalytic functions of nucleic acids20,21 has
expanded the breadth of biocatalytic protocols to include
RNA and DNA catalysts, initiating the pursuit of nucleic acid–
based enzymes. DNA possesses inherent advantages as
a catalyst. A catalytic sequence can be entirely identied from
a random sequence population and folds into its practical
tertiary structure spontaneously. A number of asymmetric
reactions, especially Lewis acid-catalyzed reactions, have
been successfully realized using DNA-based biocatalysts
resulting in remarkable performances.22–31 Recently, the
DNA catalysis is opening a promising avenue for biosyn-
thesis, but the catalytic scope and performance of DNA cata-
lysts still need improvement in comparison with catalysts
based on protein enzymes. Considering the great role played
by the cofactor ligand in the rst-coordination-sphere in
catalytic reactions,35–40 cofactor modication, which is as
powerful as directed evolution41–44 but seriously disregarded in
the eld of DNA catalysis, is introduced for the development of
DNA-based biocatalysts. Here, we report a cyclopropanation
reaction catalyzed by a G-quadruplex (G4)–Fe-porphyrin bio-
catalyst that results in enantioselectivity as high as 91%. By
tuning the N-methyl position of Fe-porphyrin from the para- to
the ortho-position, the catalytic enantioselectivity of the
reconstituted G4–Fe-porphyrin biocatalyst reverses to ꢀ72%
eetrans
. Complementary spectral, nuclear magnetic and
isothermal titration characterization studies reveal that the
stereo-divergence of the product mainly arises from the
participation of the porphyrin ligand in the regulation of
the enantioselectivity. This work succeeds in diversifying
the functionality of DNA-based biocatalysts via cofactor
modication, highlighting the great potential for cofactor
modication in the eld of DNA-based biocatalyst engineering.
aState Key Laboratory of Catalysis, Dalian Institute of Chemical Physics, Chinese
Academy of Sciences, Zhongshan Road 457, Dalian 116023, China. E-mail: canli@
dicp.ac.cn
bUniversity of Chinese Academy of Sciences, Beijing 101408, China
† Electronic supplementary information (ESI) available. See DOI:
10.1039/d1sc00755f
7918 | Chem. Sci., 2021, 12, 7918–7923
© 2021 The Author(s). Published by the Royal Society of Chemistry

View Article Online
Edge Article
Chemical Science
Table 1 Activities and selectivities of FeTMPyPn and mA9A-FeTMPyPn
(n ¼ 4, 3, 2) in the cyclopropanation reaction of styrene with EDAa
Results and discussion
Fig. 1a illustrates the design of the G4-based biocatalysts. Three
Fe-meso-tetra-(N-methylpyridyl)porphyrins with para- (FeTM-
PyP4), meta- (FeTMPyP3), and ortho- (FeTMPyP2) N-methyl
substituents were chosen as parallel cofactors to investigate the
effect of the rst-coordination-sphere on the catalytic perfor-
mance. The non-covalent binding of the cofactors with mA9A
G4 (d[G2T2G2TGAG2T2G2A]), a thrombin binding aptamer (TBA)
variant, formed the G4-based biocatalysts. The assembled bio-
catalysts were then tested in a reaction between styrene and
ethyl diazoacetate (EDA), which results in a chiral cyclopropane
product (Fig. 1b).
Table 1 lists the results obtained using the Fe-porphyrins
and their corresponding G4 biocatalysts. The use of the free
FeTMPyPn (n ¼ 4, 3, 2) cofactor as the catalyst led to relatively
low activities and no chiral induction (Table 1, entries 1–3). The
biocatalysts (mA9A-FeTMPyPn, n ¼ 4, 3, 2) assembled from
FeTMPyPn and mA9A G4 signicantly improved the catalytic
activities with turnover frequencies (TOF) increased about 10-
fold when compared to those of the free FeTMPyPns. More
surprisingly, mA9A-FeTMPyP2 induces the inversion of the
enantioselectivity relative to that of mA9A-FeTMPyP4 from
+74% to ꢀ46% (Table 1, entries 4–6). To further understand the
complementary chiral induction mechanism, circular
dichroism (CD), nuclear magnetic resonance (NMR), uores-
cence titration and other characterization methods were then
performed.
% product
conversion
b
Entry
Catalyst
TOF (hꢀ1
)
% eetrans
1
2
3
4
5
6
FeTMPyP4
FeTMPyP3
FeTMPyP2
mA9A-FeTMPyP4
mA9A-FeTMPyP3
mA9A-FeTMPyP2
5
6
4
46
36
27
16
20
11
184
144
108
0
0
0
74
63
ꢀ46
a
The experiments were carried out with styrene (30 mM), EDA (10 mM),
DNA (12.5 mM), and FeTMPyPn (n ¼ 2, 3, 4) (12.5 mM) in 10 mM
potassium phosphate buffer (pH 7.0) under argon at 4 ꢁC for 2 h,
unless otherwise specied. Product conversions and ee values are
based on the areas of HPLC peaks as compared to an internal
standard. The diastereomeric ratios of the products ranged from
90 : 10 to 97 : 3. All data are the average of 3 attempts, reproducibility
ꢂ5%. b (R,R)–(S,S).
absorb at 420 nm, but FeTMPyP2 does (Fig. 2b). Therefore, the
ICD signal can be attributed to FeTMPyP2, whose planar
symmetry is broken due to the interaction with mA9A.
The ultraviolet-visible (UV) absorption spectra (Fig. 2b) of
mA9A-FeTMPyPn further support the CD results. The spectra of
FeTMPyPn (n ¼ 4, 3, 2) exhibit broad Soret absorption bands at
around 420 nm. As the N-methyl group varies from the para- to
the ortho-position, the Soret band shows a blue shi, indicating
that FeTMPyP2 has decreased electronic conjugation relative to
the other porphyrins. Due to the steric hindrance of the 2-N-
methyl group, the bond between the pyridine ring and the
porphyrin ring of FeTMPyP2 has a tendency to rotate, causing
the pyridine ring and the porphyrin ring to be non-co- planar
(Fig. 2b, right inset), which explains the generation of an ICD
signal in mA9A-FeTMPyP2. When well-folded mA9A is added to
FeTMPyPn, the Soret bands rst fall to a minimum and then
slightly rise for all cofactors. This suggests that FeTMPyPn (n ¼
4, 3, 2) have a similar external stacking mode on mA9A, rather
than intercalating between two G-quartets.45–47 Moreover,
binding with FeTMPyP4 and FeTMPyP3 makes mA9A more
stable according to UV melting experiments (Fig. S1†), while
binding with FeTMPyP2 does not.
The CD spectra in Fig. 2a show that the antiparallel G4
signatures of mA9A, featuring two positive peaks at 245 nm and
295 nm, and one negative peak at 265 nm, are still present in the
mA9A-FeTMPyPn (n ¼ 4, 3, 2) catalysts. But of note is that the
CD spectrum of mA9A-FeTMPyP2 shows a specically induced
CD signal (ICD) at 420 nm. Only when an asymmetric confor-
mation is formed can an ICD signal be induced. DNA does not
To obtain further structural information on the mA9A-
FeTMPyPn catalysts, we conducted an NMR characterization of
the systems. The NMR spectrum of mA9A shows seven discrete
peaks over the range of 11.5–12.5 ppm (Fig. 2c), which can be
assigned to the eight guanine imino protons (H1) of the G-
quartet. These signals indicate the formation of a two-layer
antiparallel G4 structure in the potassium phosphate buffer
(Fig. 2d), like that of TBA.48,49 NMR titration experiments
(Fig. 2c) where mA9A is added to the three FeTMPyPn (n ¼ 4, 3,
2) cofactors show similar line broadening and reductions in
peak intensity. However, there are two adjacent peaks around
12.0 ppm (peaks labelled with *) with different relative declines,
indicating that the coordination modes between mA9A and the
different Fe-porphyrin cofactors vary slightly. Moreover, the
addition of FeTMPyP2 causes an obvious upeld peak-shi,
which is attributed to the strong electronic shielding effect
Fig. 1 (a) The assembly of the G4-based biocatalysts. (b) The asym-
metric cyclopropanation reaction investigated in this work.
© 2021 The Author(s). Published by the Royal Society of Chemistry
Chem. Sci., 2021, 12, 7918–7923 | 7919

View Article Online
Chemical Science
Edge Article
Fig. 2 (a) CD spectra of mA9A and mA9A-FeTMPyPn (n ¼ 4, 3, 2) (inset: a detailed view of the ICD signals). (b) UV-vis absorption spectra of
FeTMPyPn (n ¼ 4, 3, 2) when titrated with mA9A. (c) NMR spectra of mA9A over the range of 11.5–12.5 ppm when titrated with FeTMPyPn (n ¼ 4,
3, 2). (d) The G-quadruplex structure of mA9A.
arising from the porphyrin ligand stacking upon the G- separately with the uorophore 5-carboxyuorescein (FAM).
quartet.50 Therefore, in combination with the CD and UV-vis The uorescence of FAM can be suppressed by the presence of
results, we nd that FeTMPyP2 adjusts to an optimal confor- FeTMPyPn (Fig. 3a and S2†). By adding FeTMPyPn dropwise to
mation through bond rotation and distortion, thereby forming the FAM-labelled mA9A, we obtained the titration quenching
a tight p–p stacking mode with mA9A and creating a strong curves (Fig. S3†). The tted apparent equilibrium dissociation
electronic shielding effect on the imino protons of the G- constants (Kdapp) at the different FAM-labelled sites, shown in
quartet.
Fig. 3b, indicate that all the FeTMPyPn (n ¼ 4, 3, 2) cofactors
A uorescence-based binding assay was then implemented prefer binding at the 50, 30 end of the G-quartet. Considering
to locate the catalytic sites.51 Since UV titration had determined that the ratio of mA9A and FeTMPyPn is greater than 1 : 1 when
the external stacking mode of the FeTMPyPn (n ¼ 4, 3, 2) assembling the DNA-based biocatalyst and that the eetrans
cofactors on mA9A, the 50 end (50FAM-mA9A) and loop 1 values for the catalytic cyclopropanation can be maintained at
(int0FAM-mA9A) of the two G-quartets of mA9A were labelled the highest level, the preferential binding site of FeTMPyPn with
mA9A is regarded as the active center for chiral regulation.
Therefore, according to the uorescence-based binding assay,
FeTMPyPn binding at the 50, 30-end of the G-quartet of mA9A
constructs the active catalytic site of mA9A-FeTMPyPn. The
Kdapp of FeTMPyP2 is the lowest at 38 nM, compared to 70 nM
for FeTMPyP3 and 52 nM for FeTMPyP4, indicating that
FeTMPyP2 binds the strongest with mA9A.
Isothermal titration calorimetry (ITC) provides information
on intermolecular interactions by recording the heat discharged
or consumed during a bimolecular reaction. Fig. 3c shows that
there are several distinct exothermic processes during the
titration of mA9A with FeTMPyP4 and FeTMPyP3, indicating
complicated multiple binding behaviours between FeTMPyP4,
FeTMPyP3 and mA9A. As explained in the above section,
although mA9A enables the binding of multiple iron porphy-
rins, the strongest binding sites were the catalytic sites for the
chiral regulation of the cyclopropanation reaction. The titration
curves were tted using a sequential binding sites model to
Fig. 3 (a) A schematic diagram of the fluorescence quenching equi-
calculate the binding parameters. The highest affinities
librium dissociation binding assay. (b) Binding curves determined using
between FeTMPyPn and mA9A were quantied to be 33 nM (n ¼
fluorescence quenching titration. (c) ITC profiles for the binding of
4), 50 nM (n ¼ 3) and 14 nM (n ¼ 2), coinciding with the trend of
FeTMPyPn (n ¼ 4, 3, 2) with mA9A.
7920 | Chem. Sci., 2021, 12, 7918–7923
© 2021 The Author(s). Published by the Royal Society of Chemistry

View Article Online
Edge Article
Chemical Science
DNA pocket for cyclopropanation catalysis is provided by all the
mA9A-FeTMPyPn (n ¼ 4, 3, 2) catalysts (Fig. 4a). The ligands
TMPyP4 and TMPyP3 are planar symmetric molecules that are
unable to induce chirality in the catalytic process of the mA9A-
FeTMPyP4 and mA9A-FeTMPyP3 catalysts. It is the deoxy-
nucleotide residues at the 50, 30-end of the G-quartet of mA9A
that hold the iron porphyrin carbene (IPC) intermediate in
a certain orientation and dene the conguration of the
product (Fig. 4b). Nevertheless, mA9A-FeTMPyP3 shows
a similar but lower enantioselectivity than mA9A-FeTMPyP4.
Considering the similar binding strengths of FeTMPyP3 with
the two G-quartets of mA9A (Fig. 3b), the reduction in enan-
tioselectivity can be reasonably attributed to the multi-site
binding behaviour of FeTMPyP3. In contrast to mA9A-
FeTMPyP4 and mA9A-FeTMPyP3, the symmetry breaking of
the porphyrin ring in TMPyP2 allows it to participate in chiral
regulation. This, in synergy with the deoxynucleotide residues,
induces the formation of the cyclopropane product with the
opposite conguration to that catalyzed by mA9A-FeTMPyP4
(Fig. 4b).
Fig. 4 (a) A schematic diagram of the assembly of mA9A-FeTMPyPn (n
¼ 4, 3, 2). (b) Schematic diagrams of the active centres of mA9A-
FeTMPyP4 and mA9A-FeTMPyP2.
Kdapp as measured using uorescence titration. This further
supports the conclusion that the 50, 30-end of the G-quartet of
mA9A constructs the active catalytic sites. Given that FeTMPyP2
has exible peripheral groups (according to UV spectra), the
tight assembly with mA9A can be attributed to the conforma-
tional transition of the TMPyP2 ligand, which is also conrmed
by the CD spectra.
NMR, UV and uorescence titration studies indicate that
ne-tuning the N-methyl position of the cofactor does not
change the preference of FeTMPyPn for binding at the 50, 30-end
of the G-quartet of mA9A. This suggests that a similar active
To extend the substrate scope of the mA9A-FeTMPyPn cata-
lyzed cyclopropanation reaction, a series of olens and diazo-
esters were investigated (Table 2). All three mA9A-FeTMPyPn (n
¼ 4, 3, 2) catalysts show obvious substrate specicities. The
Table 2 Substrate scope for mA9A-FeTMPyPn (n ¼ 4, 3, 2) catalyzed cyclopropanationa
mA9A-FeTMPyP4
mA9A-FeTMPyP3
mA9A-FeTMPyP2
% product
% product
% product
conversion TOF (hꢀ1
)
% eetrans
conversion TOF (hꢀ1
)
% eetrans
conversion TOF (hꢀ1
)
% eetrans
b
b
b
Entry Product
1
2
3
4
5
6
1c, R ¼ 4-H
46
36
55
20
23
24
184
144
220
80
92
86
74
70
58
65
66
54
36
35
46
19
17
20
144
140
184
76
68
80
63
60
52
53
41
40
27
25
26
24
29
27
108
100
104
96
116
108
ꢀ46
ꢀ45
ꢀ46
ꢀ50
ꢀ61
ꢀ72
2c, R ¼ 4-Me
3c, R ¼ 4-OMe
4c, R ¼ 4-Cl
5c, R ¼ 4-F
6c, R ¼ 3, 4-F
7
39
156
17
35
90
10
30
120
ꢀ3
0.5
8
9
10
11
8c, R2 ¼ t-Bu
39
47
186
188
164
168
82
85
91
36
35
41
39
40
140
164
156
160
60
68
64
25
18
23
27
29
72
92
108
116
ꢀ35
ꢀ35
ꢀ31
ꢀ11
9c, R2 ¼ CH(i-Pr)2
10c, R2 ¼ CCH3(i-Pr)2 41
11c, R2 ¼ CH(Cy)2
42
a
The experiments were carried out with styrene (30 mM), EDA (10 mM), DNA (12.5 mM), and FeTMPyPn (n ¼ 2, 3, 4) (12.5 mM) in 10 mM potassium
phosphate buffer (pH 7.0) under argon at 4 ꢁC, 2 h, unless otherwise specied. Product conversions and ee values are based on the areas of HPLC
peaks compared to an internal standard. The diastereomeric ratios of the products ranged from 86 : 14 to 97 : 3. All data are the average of 3
attempts, reproducibility ꢂ5%. b (R,R)–(S,S).
© 2021 The Author(s). Published by the Royal Society of Chemistry
Chem. Sci., 2021, 12, 7918–7923 | 7921

View Article Online
Chemical Science
Edge Article
catalytic enantioselectivities of mA9A-FeTMPyP4 and mA9A- supervised the project. All authors contributed to the nal
FeTMPyP3 toward olen substrates with substituents on the manuscript.
phenyl group are reduced (entries 1 vs. 2–6). The enlargement of
the diazoester functional group from ethyl (Et) to –CCH3(i-Pr)2
(i-Pr is the abbreviation for isopropyl) enables a signicant
Conflicts of interest
enhancement in the eetrans values to 91% (entries 1 vs. 8–10).
However, the use of a –CH(Cy)2 (Cy is the abbreviation for
There are no conicts to declare.
cyclohexyl, entry 11) group does not result in an enhancement.
Several studies have reported that the modication of diazo- Acknowledgements
ester substituents causes a signicant impact on the activities
The work was supported by the National Key R&D Program of
and selectivities of carbene transfer reactions.6,52 The G4–Fe-
China (2017YFB0702800) and the Strategic Priority Research
porphyrin-catalyzed cyclopropanation has been characterized
Program of the Chinese Academy of Sciences (Grant No.
to proceed through a catalytic IPC intermediate.34 Diazoester
XDB17000000). We thank Dr Ruotian Chen for his help with
reagents attack the active [Fe] center to form an IPC interme-
revising the manuscript. We thank Xianghui Liu and Kai Wang
diate and release one molecule of N2. An appropriate substit-
for helpful discussion. We thank A/Prof. Xuanjun Ai and Dr
uent on the diazoester reagent can coordinate with the
Ting Yang for their help with the NMR and ITC experiments.
deoxynucleotide residues (especially dA9) through hydrophobic
interactions to directly determine the conformation and prop-
erties of the IPC intermediate. Therefore, a –CCH3(i-Pr)2
substituent promotes IPC convergence to a single well-dened
Notes and references
orientation, and the steric hindrance of the deoxynucleotide
residues allows one face of the IPC to be exposed to the olen,
while keeping the other inaccessible, resulting in high enan-
tioselectivity. For mA9A-FeTMPyP2, variation in the substitu-
ents on the phenyl group of the olen increases the
enantioselectivity to ꢀ72% eetrans (entry 6), but the catalytic
cyclopropanation activities are generally lower than those of the
other two biocatalysts. Although the three mA9A-FeTMPyPn (n
¼ 4, 3, 2) catalysts show different responses to substrate varia-
tion, they are all trans product-selective with trans/cis ratios of
more than 86 : 14, and almost nonselective toward the R1-
substituted olen substrate (entry 7).
1 H. Lebel, J. F. Marcoux, C. Molinaro and A. B. Charette,
Chem. Rev., 2003, 103, 977.
2 C. Ebner and E. M. Carreira, Chem. Rev., 2017, 117, 11651.
3 P. S. Coelho, Z. J. Wang, M. E. Ener, S. A. Baril, A. Kannan,
F. H. Arnold and E. M. Brustad, Nat. Chem. Biol., 2013, 9, 485.
4 P. S. Coelho, E. M. Brustad, A. Kannan and F. H. Arnold,
Science, 2013, 339, 307.
5 M. Bordeaux, V. Tyagi and R. Fasan, Angew. Chem., Int. Ed.,
2015, 54, 1744.
6 V. Tyagi and R. Fasan, Angew. Chem., Int. Ed., 2016, 55, 2512.
7 P. Bajaj, G. Sreenilayam, V. Tyagi and R. Fasan, Angew.
Chem., Int. Ed., 2016, 55, 16110.
8 A. Iffland, S. Gendreizig, P. Tafelmeyer and K. Johnsson,
Biochem. Biophys. Res. Commun., 2001, 286, 126.
9 R. Patel, Adv. Synth. Catal., 2001, 343, 527.
Conclusions
In conclusion, stereo-divergence of G4 biocatalyst-catalyzed 10 R. E. Cobb, N. Sun and H. Zhao, Methods, 2013, 60, 81.
olen cyclopropanation was achieved via cofactor modica- 11 A. Tinoco, V. Steck, V. Tyagi and R. Fasan, J. Am. Chem. Soc.,
tion. By tuning the N-methyl substituent of the porphyrin ligand
2017, 139, 5293.
in the cofactor from the para- to the ortho-position, the self- 12 A. L. Chandgude, X. K. Ren and R. Fasan, J. Am. Chem. Soc.,
assembled G4–Fe-porphyrin biocatalysts are able to switch the
enantioselectivity of the reaction from +91% to ꢀ72% eetrans
CD, NMR, ITC, and other characterization studies reveal that
2019, 141, 9145.
13 A. M. Knight, S. B. J. Kan, R. D. Lewis, O. F. Brandenberg,
K. Chen and F. H. Arnold, ACS Cent. Sci., 2018, 4, 372.
.
the porphyrin ligand cooperating with the deoxynucleotide 14 X. K. Ren, N. Y. Liu, A. L. Chandgude and R. Fasan, Angew.
residues gives the IPC intermediate a single well-dened
Chem., Int. Ed., 2020, 59, 21634.
conguration and results in a specic enantiopreference. This 15 K. Huber, B. Hamad and P. Kirkpatrick, Nat. Rev. Drug
nding is down to the rational design of DNA-based biocatalysts Discovery, 2011, 10, 255.
through cofactor modication, a method which serves as an 16 K. E. Hernandez, H. Renata, R. D. Lewis, S. B. Jennifer Kan,
effective way to regulate the catalytic performance of DNA-based
biocatalysts.
C. Zhang, J. Forte, D. Rozzell, J. A. McIntosh and
F. H. Arnold, ACS Catal., 2016, 6, 7810.
17 R. M. Atkinson and K. S. Ditman, Clin. Pharmacol. Ther.,
1965, 6, 631.
Author contributions
´
18 M. J. S. ReneCsuk and Y. von Scholz, Tetrahedron: Asymmetry,
J. Y. Hao conceived of the presented idea. J. Y. Hao, W. H. Miao,
1996, 7, 3505.
and S. M. Lu planned the experiments. J. Y. Hao and W. H. Miao 19 A. Z. Khan, M. Bilal, T. Rasheed and H. M. N. Iqbal, Chin. J.
carried out the experiments. J. Y. Hao, W. H. Miao, S. M. Lu, Y. Catal., 2018, 39, 1861.
Cheng, and C. Li contributed to the interpretation of the results. 20 S. K. Silverman, Trends Biochem. Sci., 2016, 41, 595.
J. Y. Hao, S. M. Lu, and C. Li wrote the manuscript. C. Li 21 G. F. J. Ronald and R. Breaker, Chem. Biol., 1994, 1, 223.
7922 | Chem. Sci., 2021, 12, 7918–7923
© 2021 The Author(s). Published by the Royal Society of Chemistry

View Article Online
Edge Article
Chemical Science
22 P. Fournier, R. Fiammengo and A. Jaschke, Angew. Chem., 37 S. N. Natoli and J. F. Hartwig, Acc. Chem. Res., 2019, 52, 326.
Int. Ed., 2009, 48, 4426.
23 D. Coquiere, B. L. Feringa and G. Roelfes, Angew. Chem., Int.
Ed., 2007, 46, 9308.
38 E. J. Moore, V. Steck, P. Bajaj and R. Fasan, J. Org. Chem.,
2018, 83, 7480.
39 G. Sreenilayam, E. J. Moore, V. Steck and R. Fasan, ACS
Catal., 2017, 7, 7629.
24 R. P. Megens and G. Roelfes, Chem. Commun., 2012, 48, 6366.
25 A. J. Boersma, B. L. Feringa and G. Roelfes, Angew. Chem., Int. 40 G. Sreenilayam, E. J. Moore, V. Steck and R. Fasan, Adv.
Ed., 2009, 48, 3346. Synth. Catal., 2017, 359, 2076.
26 Y. Li, M. Cheng, J. Hao, C. Wang, G. Jia and C. Li, Chem. Sci., 41 K. A. Powell, S. W. Ramer, S. B. Del Cardayre, W. P. Stemmer,
2015, 6, 5578.
M. B. Tobin, P. F. Longchamp and G. W. Huisman, Angew.
Chem., Int. Ed., 2001, 40, 3948.
42 F. H. Arnold, Acc. Chem. Res., 1998, 31, 125.
27 C. Wang, G. Jia, J. Zhou, Y. Li, Y. Liu, S. Lu and C. Li, Angew.
Chem., Int. Ed., 2012, 51, 9352.
28 M. Cheng, Y. Li, J. Zhou, G. Jia, S. M. Lu, Y. Yang and C. Li, 43 N. J. Turner, Trends Biotechnol., 2003, 21, 474.
Chem. Commun., 2016, 52, 9644. 44 G. F. Joyce, Annu. Rev. Biochem., 2004, 73, 791.
29 A. J. Boersma, R. P. Megens, B. L. Feringa and G. Roelfes, 45 C. Wei, G. Jia, J. Yuan, Z. Feng and C. Li, Biochemistry, 2006,
Chem. Soc. Rev., 2010, 39, 2083. 45, 6681.
30 A. Rioz-Martinez and G. Roelfes, Curr. Opin. Chem. Biol., 46 C. Wei, G. Jia, J. Zhou, G. Han and C. Li, Phys. Chem. Chem.
2015, 25, 80.
Phys., 2009, 11, 4025.
31 G. Roelfes, Mol. BioSyst., 2007, 3, 126.
47 C. Wei, J. Wang and M. Zhang, Biophys. Chem., 2010, 148, 51.
32 A. Rioz-Martinez, J. Oelerich, N. Segaud and G. Roelfes, 48 R. F. Macaya, P. Schultze, F. W. Smith, J. A. Roe and J. Feigon,
Angew. Chem., Int. Ed., 2016, 55, 14136. Proc. Natl. Acad. Sci. U. S. A., 1993, 90, 3745.
33 H. Ibrahim, P. Mulyk and D. Sen, ACS Omega, 2019, 4, 15280. 49 P. Schultze, R. F. Macaya and J. Feigon, J. Mol. Biol., 1994,
34 J. Y. Hao, W. H. Miao, Y. Cheng, S. M. Lu, G. Q. Jia and C. Li,
ACS Catal., 2020, 10, 6561.
35 K. Ohora, H. Meichin, L. M. Zhao, M. W. Wolf, A. Nakayama,
235, 1532.
50 C. Lin, J. Dickerhoff and D. Yang, Methods Mol. Biol., 2019,
2035, 157.
J. Hasegawa, N. Lehnert and T. Hayashi, J. Am. Chem. Soc., 51 M. P. Cheng, J. Y. Hao, Y. H. Li, Y. Cheng, G. Q. Jia, J. Zhou
2017, 139, 17265. and C. Li, Biochimie, 2018, 146, 20.
36 K. Oohora, A. Onoda and T. Hayashi, Acc. Chem. Res., 2019, 52 V. Steck, D. M. Carminati, N. R. Johnson and R. Fasan, ACS
52, 945.
Catal., 2020, 10, 10967.
© 2021 The Author(s). Published by the Royal Society of Chemistry
Chem. Sci., 2021, 12, 7918–7923 | 7923