Biocatalytical Asymmetric Sulfoxidation by Identifying Cytochrome P450 from Parvibaculum Lavamentivorans DS-1

Article abstract of DOI:10.1002/cctc.201801139

Cytochrome P450 monooxygenases (P450s) catalyzed asymmetric sulfoxidation represents a green route for the synthesis of valuable enantiopure sulfoxides, which are potentially interesting synthons in synthetic and pharmaceutical chemistry. Here the potential P450 and redox partner genes from Parvibaculum lavamentivorans DS-1 are screened and co-expressed in Escherichia coli host to construct twenty recombinant P450 strains. By testing the whole-cell biooxidation of thioanisole, P450_PL2 (CYP278A4) and P450_PL7 (CYP108G3) are identified with excellent S enantioselectivity while P450_PL1 (CYP111B1) and P450_PL9 (CYP153A26) exhibit the complementary R enantioselectivity. Asymmetric sulfoxidation of sulfides 1 a–1 m is further investigated using the recombinant E. coli strain P450_PL2-2 based on the optimal conditions, producing the corresponding enantioenriched sulfoxides with up to 82 % isolated yield and 99 % ee.

Full text of DOI:10.1002/cctc.201801139

Accepted Article
Title: Biocatalytical asymmetric sulfoxidation by identifying cytochrome
P450 from Parvibaculum lavamentivorans DS-1
Authors: Kailin Wu, Linchao Tang, Haibo Cui, Nanwei Wan, Ziyan Liu,
Zhongqiang Wang, Shimin Zhang, Baodong Cui, Wenyong
Han, and Yongzheng Chen
This manuscript has been accepted after peer review and appears as an
Accepted Article online prior to editing, proofing, and formal publication
of the final Version of Record (VoR). This work is currently citable by
using the Digital Object Identifier (DOI) given below. The VoR will be
published online in Early View as soon as possible and may be different
to this Accepted Article as a result of editing. Readers should obtain
the VoR from the journal website shown below when it is published
to ensure accuracy of information. The authors are responsible for the
content of this Accepted Article.
To be cited as: ChemCatChem 10.1002/cctc.201801139
Link to VoR: http://dx.doi.org/10.1002/cctc.201801139
A Journal of
www.chemcatchem.org

ChemCatChem
10.1002/cctc.201801139
COMMUNICATION
Biocatalytical asymmetric sulfoxidation by identifying
cytochrome P450 from Parvibaculum lavamentivorans DS-1
[
a]
[a]
[a]
[a]
[a]
[a]
Kailin Wu, ‡ Linchao Tang, ‡ Haibo Cui, Nanwei Wan, * Ziyan Liu, Zhongqiang Wang, Shimin
[
a]
[a]
[a]
[a]
Zhang, Baodong Cui, Wenyong Han, Yongzheng Chen *
Abstract: Cytochrome P450 monooxygenases (P450s) catalyzed
asymmetric sulfoxidation represents a green route for the synthesis
of valuable enantiopure sulfoxides, which are potentially interesting
synthons in synthetic and pharmaceutical chemistry. Here the
potential P450 and redox partner genes from Parvibaculum
lavamentivorans DS-1 are screened and co-expressed in
Escherichia coli host to construct twenty recombinant P450 strains.
By testing the whole-cell biooxidation of thioanisole, P450PL2
challenge to seek promising P450s for asymmetric sulfoxidation
and expand the substrate scope.
Our research focused on the enzymatic oxidation
reactions,[15] including high regio- and stereo-selective
biocatalytic asymmetric sulfoxidation, and have developed a
stereoselective oxidation method to synthesize optically active
sulfoxides with resting cells of Pseudomonas monteilii CCTCC
M2013683.[16] In this study, we tried to identify and
heterogeneously express nine potential P450s and five redox
partners genes from P. lavamentivorans DS-1. After evaluating
the activity and enantioselectivity of twenty recombinant P450
strains toward thioanisole, strain P450PL2-2 was selected and
optimized for asymmetric sulfoxidation of sulfides 1a-1m,
resulting in the formation of the corresponding enantiomerically
pure sulfoxide derivatives with up to 82% isolated yield and 99%
ee.
(
CYP274A4) and P450PL7 (CYP108G3) are indentified with excellent
enantioselectivity while P450PL1 (CYP111B) and P450PL9
CYP153A26) exhibit the complementary enantioselectivity.
S
(
R
Asymmetric sulfoxidation of sulfides 1a-1m is further investigated
using the recombinant E. coli strain P450PL2-2 based on the optimal
conditions, producing the corresponding enantioenriched sulfoxides
with up to 82% isolated yield and 99% ee.
We first searched the genome sequence of the strain P.
lavamentivorans DS-1, and identified a total of nine potential
P450 gene sequences (P450PL1-P450PL9). All the P450s belong
to Class I family. According to the P450 classification proposed
by Nelson,[17] P450PL1(CYP111B1) is the only member of the
CYP111B family, while there are more than 100 sequences with
The cytochrome P450 monooxygenases (P450s or CYPs) are
versatile biocatalysts that ubiquitously exist in almost all living
systems,[1] and involved in a diverse range of oxidation reactions
[
2]
[3]
[4]
such as hydroxylation, epoxidation, sulfoxidation, oxidative
phenolic coupling,[5] N- and O-dealkylation, carbon–carbon
bond cleavage[7] and cyclopropanation.[8] The bacterial P450s
can be divided into ten classes according to their topologies.[9]
The Class I P450s consist of a ferredoxin reductase (Fdr) and a
ferredoxin (Fdx) that shuttle electrons from the cofactor
NAD(P)H to the monooxygenase component, such as P450cam
[6]
>40% identity to P450PL1 being identified to date.
P450PL2(CYP278A4) belongs to the CYP278A family. The other
three closet homologs are present in Mycobacterium marinum
(
CYP278A1), Mycobacterium vanbaalenii PYR-1 (CYP278A2)
and Ectocarpus bacterium (CYP278A3), but none of them has
been characterized. P450PL7(CYP108G3) is a member of the
CYP108G family. P450PL8(CYP1010A1) is classified in
CYP1010A family, and blast search reveals another P450
(
CYP101A1)[10] and P450pyr (CYP153A7)[11]
Chiral sulfoxides widely exist in many biologically active
.
molecules and play an important role in asymmetric
transformations.[12] Hence, practical synthetic methods for the
chiral sulfoxides have constantly been an interesting subject
over the past two decades. The most direct approach is
asymmetric oxidation of the prochiral sulfides, which is
undoubtedly a real breakthrough in this area and has been well
documented.[13] Among them, the P450 catalyzed sulfoxidation
represents a green and attractive route in the synthesis of chiral
sulfoxide due to its high stereoselectivity and mild condition.
However, only several P450s has been studied on asymmetric
(
WP_052343946.1) from strain Kallotenue papyrolyticum JKG1
that shares 41% similarity to P450PL8. The other five P450s
including P450PL3(CYP153A31), P450PL4(CYP153A30),
P450PL5(CYP153A29), P450PL6(CYP153A27) and
P450PL9/CYP153A26) originate from the CYP153 family. For this
P450 family, several members have been isolated for
hydroxylation of alkanes and fatty acids such as n-alkane and
(
S)-limonene.[18]
All the above P450 genes were successfully amplified,
sulfoxidation with limited substrates.[14] Thus, it is still
a
except for P450PL4
.
The amplified P450 genes were
heterologously expressed in E. coli BL21(DE3) strain with
pET28a-(+) vector. SDS-PAGE analysis showed production of
soluble P450PL1, P450PL2 and P450PL7 was achieved. SDS-
PAGE analysis also showed that P450PL3 and P450PL9
generated inclusion body. Further work achieved the expression
of soluble P450PL9 by using another vector, the pET32a-(+).
However, application of this method to the P450PL3 was
unsuccessful. Besides, no obvious expression band
corresponding to P450PL5, P450PL6 and P450PL8 was found. The
protein expression and carbon monoxide (CO) difference
spectra results of the P450PL1, P450PL2, P450PL7 and P450PL9
[
a]
K. Wu, L. Tang, H. Cui, Dr. N. Wan, Z. Liu, Dr. Z. Wang, Dr. S.
Zhang, Dr. B. Cui, Dr. W. Han, Prof. Dr. Y. Chen
Generic Drug Research Center of Guizhou Province, Green
Pharmaceuticals Engineering Research Center of Guizhou
Province, School of Pharmacy, Zunyi Medical University, Zunyi,
563000, China.
Dr. Nanwei Wan*, E-mail: nanweiwan@zmc.edu.cn.
Prof. Dr. Yongzheng Chen*, E-mail: yzchen@zmc.edu.cn.
These authors contributed equally
[‡]
Supporting information for this article is given via a link at the end of
the document.((Please delete this text if not appropriate))
This article is protected by copyright. All rights reserved.

ChemCatChem
10.1002/cctc.201801139
COMMUNICATION
were shown in Figure S1 and S2, which indicated P450PL2 and
P450PL7 were properly folded in the recombinant cells. However,
the CO difference spectra at 450 nm of P450PL1 and P450PL9
were not distinct. Subsequently, one Fdr gene and five Fdx
genes (Fdx1, Fdx2, Fdx3, Fdx4 and Fdx5) were successfully
amplified and inserted into the CDFDuet-1 plasmid to construct
five co-expressed plasmids CDFDuet-Fdr-Fdx. The co-
expression of Fdr and Fdx genes shown in Figure S3 indicated
the successful expression of the five different electron transport
systems. Subsequently, the recombinant plasmids pET28-
P450PL1, pET28-P450PL2, pET28-P450PL7 and pET32-P450PL9
were transformed to E. coli BL21(DE3) together with five
recombinant redox partner plasmids (CDFDuet-Fdr-Fdx),
respectively. Co-expression of P450 component and redox
partners were confirmed by SDS-PAGE analysis. The results
displayed in Figure 1 revealed that co-expression of Fdr-Fdxs
with the four P450s were achieved, and twenty recombinant
P450 strains containing the three complete components were
constructed.
the activities of the class I P450s were alterable by changing the
redox partner system.
Table 1. Asymmtric sulfoxidation of 1a using the recombinant P450 strains.
Redox
partner
P450
strains
Entry[a]
Yield (%)[b]
ee (%)[b]
Config.[b]
N.D.[c]
1
2
3
4
5
6
7
8
9
Fdr-Fdx1
Fdr-Fdx2
Fdr-Fdx3
Fdr-Fdx4
Fdr-Fdx5
Fdr-Fdx1
Fdr-Fdx2
Fdr-Fdx3
Fdr-Fdx4
Fdr-Fdx5
Fdr-Fdx1
Fdr-Fdx2
Fdr-Fdx3
Fdr-Fdx4
Fdr-Fdx5
Fdr-Fdx1
Fdr-Fdx2
Fdr-Fdx3
Fdr-Fdx4
Fdr-Fdx5
P450PL1-1
P450PL1-2
P450PL1-3
P450PL1-4
P450PL1-5
P450PL2-1
P450PL2-2
P450PL2-3
P450PL2-4
P450PL2-5
P450PL7-1
P450PL7-2
P450PL7-3
P450PL7-4
P450PL7-5
P450PL9-1
P450PL9-2
P450PL9-3
P450PL9-4
P450PL9-5
<5
14
6
5
8
N.D.
R
R
R
R
S
S
S
S
S
89
70
83
86
95
96
95
95
85
96
95
97
93
95
84
91
76
92
70
46
48
35
43
21
43
44
40
45
31
20
21
20
38
7
1
1
0
1
S
S
S
S
12
13
14
15
16
17
18
19
20
S
R
R
R
R
R
[
a] Unless otherwise noted, all reactions were performed in 5 mL KH
2 4
PO -
Na HPO
2
4
buffer (50 mM, pH 7.0) at 30 oC, 250 rpm for 24 h. All reactions were
repeated for three times. [b] The yield, ee and absolute configuration of 2a
were determined by chiral HPLC analysis and calculated by comparing with
the standard compounds. [c] N.D. = Not Determined.
Subsequently, the strain P450PL2-2 (co-expression of P450PL2
with Fdr-Fdx2) with highest activity and enantioselectiviy
towards the conversion of 1a was selected for further study, by
optimizing the reaction conditions used for the production of 2a
from 1a. The reaction results were summarized in Table 2.
Increase of the cell concentration gave no significant
improvement on the yield and enantioselectivity (entries1-3).
However, the yield reduced dramatically by increasing the
substrate concentration (entry 4), suggesting that 1a inhibits the
activity of P450PL2-2. This substrate inhibition behavior has been
found in other P450-catalyzed sulfoxidation reactions, which can
Figure 1. SDS-PAGE analysis of the co-expression of P450PL1(A), P450PL2(B),
P450PL7(C), and P450PL9(D) together with five Fdr-Fdx redox partner systems,
respectively. Lane M: standard protein marker; Lane 1: P450 co-expressed
with Fdr-Fdx1; Lane 2: P450 co-expressed with Fdr-Fdx2; Lane 3: P450 co-
expressed with Fdr-Fdx3; Lane 4: P450 co-expressed with Fdr-Fdx4; Lane 5:
P450 co-expressed with Fdr-Fdx5, Lane 6: E. coli BL21(DE3) containing
CDFDuet-1 plasmid (negative control).
Thioanisole (1a) was used as a template substrate to
determine the sulfoxidation activity of the twenty recombinant
P450 strains. The whole-cell biocatalytic reactions revealed that
all the four P450s catalyzed the conversion of 1a to 2a, as
depicted in Table 1. Strains P450PL2 (entries 6-10) and P450PL7
be remedied slightly by using aqueous/ionic liquid biphasic
system.[4] Excitingly, the yield of the compound 2a was improved
to 59% with the enantioselectivity remaining unchanged when
using ethanol as a cosolvent, which might attribute to the
increase of the solubility of 1a (entry 5). Comparing with the
weak alkaline and weak acid environment, the strain P450PL2-2
seemingly preferred neutral reaction conditions in view of the
output of 2a (entries 6 and 7 vs entry 5). Finally, change of the
reaction temperature reduced the production of 2a (entry 7).
(
entries 11-15) generated the (S)-2a, while P450PL1 (entries 1-5)
and P450PL9 (entries 16-20) prefer to produce 2a with R
enatiomer. By combing with the same redox partner, P450PL2
and P450PL7 exhibited the higher catalytic activity than P450PL1
and P450PL9 (entries 7 and 12 vs entries 2 and 17), which
accorded with the results of carbon monoxide difference spectra.
It was also found that, in some cases, different catalytic activities
were observed to the same P450 when combining with different
redox partners (entry 7 vs 10, entry 16 vs 20). The sulfoxidation
reaction did not occur when using the recombinant strains that
without coupling with the Fdr-Fdx redox partner (data not shown).
Taken together, these results brought us to the speculation that
Table 2. Optimizing reaction conditions of the strain P450PL2-2 catalysed
asymmetric sulfoxidation of 1a.
Entry[a]
pH
Substrate
Cell density
Yield
ee
This article is protected by copyright. All rights reserved.

ChemCatChem
10.1002/cctc.201801139
COMMUNICATION
-
1
(%)[b]
48
48
42
15
59
10
17
(%)[b]
96
98
99
97
98
99
97
Conc.(mM)
(dcw g L )
and gave the chiral sulfoxide 2i with 90% ee (entry 1). The good
activity was also found for 3-ketobutyl and 2-chloroethyl
substituted sulfides 1j and 1k, generating the enantiomerically
pure sulfoxides 2j and 2k with >99% ee (entries 2 and 3).
Replacing the ethyl group of sulfide 1i with an allyl group
produced nearly racemic sulfoxide 2l, with the yield being even
higher (entry 4). Furthermore, the β-ketosulfide 1m was also well
behaved in this enantioselective sulfoxidation process,
producing the corresponding chiral β-ketosulfoxide 2m with 64%
yield and 95% ee (entry 5).
1
2
3
4
7
7
7
7
7
5
9
7
2
2
2
4
2
2
2
2
10
20
30
10
10
10
10
10
[
c]
5
[
d]
d]
6
7
[
8[
d][e]
44
97
[
(
a] Unless otherwise noted, all reactions were performed in 5 mL PBS buffer
50 mM, pH 7.0) containing 2-4 mM 1a (bold 1a) at 25 oC, 250 rpm for 24 h. [b]
Determined by chiral HPLC analysis. [c] 0.4% (v/v) EtOH was used as
cosolvent. [d] The reaction was performed for 8 h. [e] Reaction was performed
o
at 30 C.
Table 4. Substrate scope of sulfide derivatives 1i-1m.
After the establishment of the reaction conditions (Table 2,
entry 5), the substrate scope of the recombinant strain P450PL2-2
mediated asymmetric sulfoxidation of aromatic sulfides 1 was
further explored. It was found that the steric property of the
substituent on the phenyl ring of the thioanisoles 1 has a
remarkable influence on the stereoselectivity of the oxidation
reaction. As indicated in Table 3, the thioanisoles 1b-c
containing a methyl substituent on the phenyl ring are readily
applied to the asymmetric sulfoxidation process, leading to the
production of the corresponding chiral sulfoxides 2b-c with
moderate degree of enantioselectivity (entries 2 and 3). By
comparison, a higher degree of enantioselectivity was obtained
by employing the thioanisoles 1d-g with a fluorine or chlorine
substituent on the phenyl ring (entries 2-7, for 2d-g, up to 72%
yield and 95% ee). The substituent at meta-position always
exhibited higher enantioselectivity than para-position (entry 2 vs
Substrate[a]
Yield (%)[b]
ee (%)[c]
Entry
1
Product
73
90
(1i)
(
2i)
2j)
2
3
4
5
60
70
82
64
99
99
17
95
(1j)
(
(1k)
(2k)
(1l)
(2l)
3, entry 5 vs 6). Notably, the 2-thienyl-substituted sulfide 1h was
also applicable to this biocatalytic oxidation process under the
optimal reaction conditions and provided the corresponding
chiral sulfoxide 2h with 49% yield and 85% ee (entry 8). It was
emphasized that the absolute configuration of the synthetic
chiral sulfoxides 2 was determined by comparison with the chiral
HPLC analysis of commercially available 2c.
(1m)
(2m)
[
(
a] Unless otherwise noted, all reactions were performed in 5 mL PBS buffer
50 mM, pH 7.0) containing 2 mM 1i-1m and 10 dcw g L cells at 25 C, 250
rpm for 8 h. 0.4% (v/v) EtOH was used as cosolvent. [b] Isolated yields. [c]
Determined by the chiral HPLC analysis.
-
1
o
Table 3. Substrate scope of sulfide derivatives 1a-1h.
Due to a number of limitations such as instability and low
activity, the application of the P450s in academic and industrial
arena has been challenging, despite it having displayed great
synthetic potential.[19] Many efforts have been made to generate
valuable non-natural activities, regioselectivity, stereoselectivity
and other properties of P450s.[20] Though the generation of
Yield
ee
Entry
Substrate[a]
Product
%)[
59
66
59
53
59
72
67
49
b]
(%)
98
77
62
80
95
82
95
85
[c]
(
1
2
3
4
5
6
7
8
Ar = Ph (1a)
2a
2b
2c
2d
2e
2f
enzyme variants by directed evolution or rational design has
been particularly popular and widely used to improve the
enzyme properties,[21] it is still a challenge to engineer the P450s,
because it is difficult to develop an effective and sensitive high
throughput screening method for the P450-catalyzed reactions,
as well as the uncoupling of the electron transport and formation
of product.
Ar = 3-MeC
Ar = 4-MeC
6
H
4
(1b)
(1c)
(1d)
(1e)
(1f)
6 4
H
Ar = 2-ClC
6 4
H
Ar = 3-ClC
6
H
4
Ar = 4-ClC
Ar = 4-FC
Ar = 2-thienyl (1h)
6 4
H
6
H
4
(1g)
2g
2h
[
(
a] Unless otherwise noted, all reactions were performed in 5 mL PBS buffer
50 mM, pH 7.0) containing 2 mM 1a-1h and 10 dcw g L strain P450PL2-2
cells at 25 C, 250 rpm for 8 h. 0.4% (v/v) EtOH was used as cosolvent. [b]
Isolated yields. [c] Determined by chiral HPLC analysis.
To date, P450 mediated asymmetric sulfoxidation has been
limited for wide application in organic synthesis. Only the
P450cam, CYP154H1, P450SMO, P450RhF, P450BM3 and
P450pyr have been studied for asymmetric sulfoxidation.[
Usually, these reactions were restricted to peform in low
substrate concentration due to the toxic and inhibitory cauased
by sulfide and sulfoxide. For instance, the P450SMO,[14b]
P450RhF[14d] and P450BM3 variants [14e] were carried out with 1-
-
1
o
14]
In order to further explore the substrate scope of the strain
P450PL2-2, some other structurally modified aromatic sulfides
were also investigated in this work (Table 4). Firstly, catalysis of
the ethyl substituted sulfides 1i showed good reaction activity
This article is protected by copyright. All rights reserved.

ChemCatChem
10.1002/cctc.201801139
COMMUNICATION
2
mM sulfides. Though P450cam[14a] and CYP154H1[14c] were
[7]
M. J. Cryle, J. J. De Voss, Chem.Commun. 2004, 35, 86-87.
P. S. Coelho, E. M. Brustad, A. Kannan, F. H. Arnold, Science 2013,
39, 307-310.
[
[
[
8]
reported to act on 4-5 mM sulfides, both gave relatively low
conversion and stereoselectivity unfortunately. By introduing
aqueous/ionic liquid biphasic system and co-expression of GDH
for co-factor regeneration, P450pyr could tolerate 30-70 mM
sulfides, however, generated low anlytical yields (23-61%) of
sulfoxide.[ More importantly, all these P450s were studied with
a narraw substrate scope (5-6 sulfides).[14] In this study, we
identified four novel P450s for asymmetric sulfoxidation with R
3
9]
F. Hannemann, A. Bichet, K. M. Ewen, R. Bernhardt, (BBA)-General
Subjects. 2007, 1770, 330-344.
10] I. Schlichting, J. Berendzen, K. Chu, A. M. Stock, S. A. Maves, D. E.
Benson, R. M. Sweet, D. Ringe, G. A. Petsko, S. G. Sligar, Science
4]
2000, 287, 1615-1622.
[11] W.-L. Tang, Z. Li, H.-M. Zhao, Chem. Commun. 2010, 46, 5461-5463.
[12] a) R. Kubec, M. Svobodová, J. Velíšek, J. Agric. Food. Chem. 2000, 48,
4
5
4
28-433; b) W. Komatsu, Y. Miura, K. Yagasaki, Lipids. 1998, 33, 499-
03; c) K. H. Kyung, D. C. Han, H. P. Fleming, J. Food Sci. 1997, 62,
06-409; d) M. J. Ashton, A. W. Bridge, R. C. Bush, D. I. Dron, N. V.
or
S enantioselectivity. In addition, we investigated the
asymmetric sulfoxidation on a more broad substrate scope and
evaluated these reactions with isolated yields of the
corresponding sulfoxides.
In conclusion, four enantioselective P450PL1 (CYP111B1),
P450PL2 (CYP278A4), P450PL7 (CYP108G3) and P450PL9
Harris, G. D. Jones, D. J. Lythgoe, Ridell, D. Smith, C. Bioorg. Med.
Chem. Lett. 1992, 2, 375–380; e) M. P. Ortega, M. C. Garcia, M. A.
Gijon, M. F. D. Casa-Juana, J. G. Priego, M. S. Crespo, C. Sunkel, J.
Pharmacol. Exp. Ther. 1990, 255, 28–33; f) M. C. Carreno, Chem. Rev.
1995, 95, 1717-1760.
(
CYP153A26) were identified and co-expressed with five
different Fdr-Fdx electron transport systems in E. coli BL21(DE3) [13] a) G. Jindal, R. B. Sunoj, Angew. Chem. Int. Ed. 2014, 53, 4432-4436;
b) Z.-G. Zhang, R. Lonsdale, J. Sanchis, M. T. Reetz, J. Am. Chem.
host. The recombinant E. coli strain P450PL2-2 was investigated
to catalyze the asymmetric sulfoxidation of the structurally
Soc. 2014, 136, 17262-17272; c) R. Feingersch, J. Shainsky, T. K.
Wood, A. Fishman, Appl. Environ. Microbiol. 2008, 74, 1555-1566; d) H.
diverse prochiral sulfides, affording a range of enantiomerically
Srour, P. Le Maux, S. Chevance, G. Simonneaux, Coord. Chem. Rev.
pure sulfoxide derivatives with up to 82% isolated yield and 99%
ee. Studies focusing on the further biocatalytic applications of
these P450s are in progress.
2013, 257, 3030-3050; e) C.-F. Zhu, X. Chen, Z.-W. Yang, X. Du, Y. Liu,
Y. Cui, Chem. Commun. 2013, 49, 7120-7122; f) T. Matsui, Y.
Dekishima, M. Ueda, Appl. Microbiol. Biotechnol. 2014, 98, 7699-7706;
g) E. Wojaczyńska, J. Wojaczyński, Chem. Rev. 2010, 110, 4303-4356;
h) J. Han, V. A. Soloshonok, K. D. Klika, J. Drabowiczef, A. Wzorek,
Chem. Soc. Rev. 2018, 47, 1307-1350.
Acknowledgements
[
14] a) J. Fruetel, Y.-T. Chang, J. Collins, G. Loew, P. R. O. D. Montellano,
J. Am. Chem. Soc. 1994, 116, 11643-11648; b) J.-D. Zhang, A.-T. Li, Y.
Yang, J.-H. Xu, Appl. Microbiol. Biotechnol. 2010, 85, 615-624; c) A.
Schallmey, G. den Besten, I. G. P. Teune, R. F. Kembaren, D. B.
Janssen, Appl. Microbiol. Biotechnol. 2011, 89, 1475-1485.; d) E.
O'Reilly, M. Corbett, S. Hussain, P. P. Kelly, D. Richardson, S. L.
Flitsch, N. J. Turner, Catal. Sci. Technol. 2013, 3, 1490-1492.; e) J.-B.
Wang, A. I, M. T. Reetz, Adv. Synth. Catal. 2017, 359, 2056-2060.
15] a) D.-J. Zheng, M. Yang, J.-R. Zhuo, K. Li, H.-Y. Zhang, J.-W. Yang, B.-
D. Cui, Y.-Z. Chen, J. Mol. Catal. B: Enzym. 2014, 110, 87-91; b) J.
Zhao, S.-C. Guan, X.-J. Zhou, W.-Y. Han, B.-D. Cui, Y.-Z. Chen,
Tetrahedron 2016, 72, 3098-3104; c) D.-J. Zheng, X.-J. Zhou, B.-D. Cui,
W.-Y. Han, N.-W. Wan, Y.-Z. Chen, ChemCatChem. 2017, 9, 937-940.
16] Y.-Z. Chen, J.-R. Zhuo, D.-J. Zheng, S. Tian, Z. Li, J. Mol. Catal. B:
Enzym. 2014, 106, 100-104.
We are grateful for financial support from National Natural
Science Foundation of China (No. 21562054 and No. 21662050),
Guizhou Science and Technology Department (QKHRC-2016-
4
029, QKHPTRC-2016-5801, and QKHRCTD-2014-4002),
Guizhou Education Department (QSHZZ-2012-169, GNYL-
017-006), The Fifth Batch of Talent Base in Guizhou Province
2016), Program for Outstanding Youth of Zunyi Medical
2
(
[
[
University (17zy-001).
Keywords: Biocatalysis • asymmetric sulfoxidation • P450
monooxygenases • enantiopure sulfoxide
[17] D. R. Nelson, Hum. Genomics 2009, 4, 59-65.
[
1]
For reviews see: a) V. B.Urlacher, S. Eiben, Trends. Biotechnol. 2006,
[
18] a) E. G. Funhoff, U. Bauer, I. García-Rubio, B. Witholt, J. B. van Beilen,
J. Bacteriol. 2006, 188, 5220-5227; b) E. G. Funhoff, J. Salzmann, U.
Bauer, B. Witholt, J. B. van Beilen, Enzyme Microb. Technol. 2007, 40,
24, 324-330; b) S. T. Jung,R. Lauchli, F. H. Arnold, Curr. Opin. Biotech.
2011, 22, 809-817; c)V. B. Urlacher, M. Girhard, Trends. Biotechnol.
2012, 30, 26-36.
8
06-812.
19] E. O'Reilly, V. Kohler, S. L. Flitsch, N. J. Turner, Chem. Commun. 2011,
7, 2490-2501.
20] a) S. T. Jung, R. Lauchli, F. H. Arnold, Curr. Opin. Biotechnol. 2011, 22,
[
2]
a) Y. Yang, Y. T. Chi, H. H. Toh, Z. Li, Chem. Commun. 2015, 51, 914-
[
[
917; b) S. Yang, Y. Wo, M. E. Meyerhoff, Anal. Chim. Acta. 2014, 843,
4
89-96.
[
[
[
[
3]
4]
5]
6]
A.-T. Li, S.-K. Wu, J. P. Adams, R. Snajdrova, Z. Li, Chem. Commun.
014, 50, 8771-8774.
P.-F. Gao, A.-T. Li, H. H. Lee, D. I. C. Wang, Z. Li, ACS Catal. 2014, 4,
763-3771.
8
09-817; b) V. B. Urlacher, M. Girhard, Trends. Biotechnol. 2012, 30,
6-36; c) J. A. McIntosh, C. C. Farwell, F. H. Arnold, Curr. Opin. Chem.
2
2
Biol. 2014, 19, 126-134.
3
[
21] U. T. Bornscheuer, M. Pohl, Curr. Opin. Chem. Biol. 2001, 5, 137-143.
K. Zerbe, K. Woithe, D.-B. Li, F. Vitali, L. Bigler, J. A. Robinson, Angew.
Chem. Int. Ed. 2004, 43, 6709-6713.
a) M. N. Bhakta, K. Wimalasena, J. Am. Chem. Soc. 2002, 124, 1844-
1845; b) S. H. Lee, Y. C. Kwon, D. M. Kim, C. B. Park, Biotechnol.
Bioeng. 2013, 110, 383-390.
This article is protected by copyright. All rights reserved.

ChemCatChem
10.1002/cctc.201801139
COMMUNICATION
Entry for the Table of Contents (Please choose one layout)
Layout 1:
COMMUNICATION
Four enantioselective P450s from
Parvibaculum lavamentivorans DS-1
were identified and co-expressed with
five redox partners to construct twenty
recombinant P450 strains. P450PL2-2
strain were optimized to develop a
green process for the synthesis of
enantiopure sulfoxides via asymmetric
sulfoxidation of prochiral sulfides,
giving the corresponding
Kailin Wu, Linchao Tang, Haibo Cui,
Nanwei Wan,* Ziyan Liu, Zhongqiang
Wang, Shimin Zhang, Baodong Cui,
Wenyong Han, Yongzheng Chen*
Page No. – Page No.
Biocatalytical asymmetric
sulfoxidation by identifying
cytochrome P450 from Parvibaculum
lavamentivorans DS-1
enantioenriched sulfoxides with up to
82% isolated yield and 99% ee.
This article is protected by copyright. All rights reserved.