ChemCatChem
10.1002/cctc.201801613
FULL PAPER
o
Am. Chem. Soc 2013, 135, 12480-12496; c) J. Dong, E. Fernández-
Fueyo, F. Hollmann, E. Paul Caroline, M. Pesic, S. Schmidt, Y.
Wang, S. Younes, W. Zhang, Angew. Chem., Int. Ed. 2018, 57,
mixture. The reaction was performed at 30 C and 250 rpm for 6 h. At the
end of the reaction, XAD4 resins were separated and washed with 2×10
mL of 99% ethanol for 2 h. 200 µL of aqueous phase was diluted with 500
µL ACN containing 2mM benzyl alcohol and 300 µL ultrapure water, while
9238-9261; d) J. H. Schrittwieser, S. Velikogne, M. Hall, W. Kroutil,
Chem. Rev. 2018, 118, 270-348; e) M. Winkler, M. Geier, P. Hanlon
Steven, B. Nidetzky, A. Glieder, Angew. Chem., Int. Ed. 2018, 57,
5
0 µL of organic phase was added to 950 µL ethyl acetate containing 2mM
1
3406-13423; f) G. Hughes, J. C. Lewis, Chem. Rev. 2018, 118, 1-
g) S. Gandomkar, A. Żądło-Dobrowolska, W. Kroutil,
benzyl alcohol. The aqueous samples were analyzed by HPLC to quantify
the concentration of L-Phe 1, trans-cinnamic acid 2, and 2-PE 6, while the
non-aqueous samples were analyzed by GC to quantify the concentration
of styrene 3, (S)-styrene oxide 4, phenylacetaldehyde 5, and 2-PE 6. The
results are given in Figure 4c.
3;
ChemCatChem 2018, 10, 1-20; h) S. Wu, Z. Li, ChemCatChem
2017, 10, 2164-2178; i) Y. Zhou, S. Wu, J. Mao, Z. Li,
ChemSusChem 2018, 11, 2221-2228.
[
2]
a) J. Schrader, M. M. W. Etschmann, D. Sell, J.-M. Hilmer, J.
Rabenhorst, Biotechnol. Lett. 2004, 26, 463-472; b) S. Serra, C.
Fuganti, E. Brenna, Trends Biotechnol. 2005, 23, 193-198.
a) D. L. Hua, X. H. Liang, C. C. Che, X. D. Zhang, J. Zhang, Y. Li,
P. Xu, Asian J. Chem 2013, 25, 5951-5954; b) P. Okuniewska, U.
Domańska-Żelazna, A. Pobudkowska-Mirecka, J. Mierzejewska,
CHEMIK 2016, 70, 491-496; c) E. Fabre Cathy, J. Blanc Philippe,
G. Goma, Biotechnol. Prog. 2008, 14, 270-274.
Biotransformation of L-Phe 1 to 2-PE 6 with Recycling of E. coli (RE)
Cells in a Biodiesel-Aqueous Two-Phase System in the Presence of
XAD4 Resins
[3]
[
[
4]
5]
M. Suástegui, Z. Shao, J. Ind. Microbiol. Biotechnol. 2016, 43, 1611-
E. coli (RE) cells (15 g cdw/L) in 10 mL KP buffer (100 mM, pH 8.0, 2%
glucose) containing 100 mM L-Phe 1 were mixed with 10 mL of biodiesel
and 1.8 g XAD4 resins. The reaction was performed at 30 C and 250 rpm
1624.
X. Tian, R. Ye, J. Wang, Y. Chen, B. Cai, S. Guan, S. Rong, Q. Li,
Electron. J. Biotechnol. 2015, 18, 286-290.
o
for 6 h. At the end of the reaction, XAD4 resins were separated and
washed with 2×10 mL of 99% ethanol for 2 h, while the cells were
harvested by centrifugation (14000 g, 5 min) and resuspended back in
fresh KP buffer. New XAD4 resins and fresh biodiesel were added into the
new mixture to perform another batch of the same biotransformation.
Sample preparation and analysis were performed in the same way as
described above.
[6]
7]
M. Etschmann, W. Bluemke, D. Sell, J. Schrader, Appl. Microbiol.
Biotechnol. 2002, 59, 1-8.
a) Z. Wang, X. Bai, X. Guo, X. He, J. Ind. Microbiol. Biotechnol.
[
2017, 44, 129-139; b) D. Hua, P. Xu, Biotechnol. Adv. 2011, 29, 654-
660.
[8]
a) D. Stark, D. Zala, T. Münch, B. Sonnleitner, I. W. Marison, U. von
Stockar, Enzyme Microb. Technol. 2003, 32, 212-223; b) M. M. W.
Etschmann, D. Sell, J. Schrader, J. Mol. Catal. B Enzym. 2004, 29,
1
87-193.
a) Z. Kang, C. Zhang, G. Du, J. Chen, Appl. Biochem. Biotechnol.
014, 172, 2012-2021; b) D. Koma, H. Yamanaka, K. Moriyoshi, T.
[
9]
2
Preparative Biotransformation of L-Phe 1 with E. coli (RE) Cells for
the Production of 2-PE 6
Ohmoto, K. Sakai, Appl. Environ. Microbiol. 2012, 78, 6203-6216; c)
S. Atsumi, T. Hanai, J. C. Liao, Nature 2008, 451, 86-89; d) D. Guo,
L. Zhang, S. Kong, Z. Liu, X. Li, H. Pan, J. Agric. Food Chem. 2018,
6
6, 5886-5891.
S. Machas Michael, R. McKenna, R. Nielsen David, Biotechnol. J.
017, 12, 1700310.
E. coli (RE) cells (10 g cdw/L) in 30 mL KP buffer (100 mM, pH 8.0, 2%
glucose) containing 1.5 mmol of L-Phe 1 were mixed with 30 mL n-
hexadecane and 2.7 g XAD4 resins. The reaction was performed at 30 C
and 250 rpm for 6 h. At the end of the biotransformation, resins were
separated and washed with 2×30 mL of 99% ethanol for 2 h. Cells were
discarded by centrifugation (4000 g, 10 min). The organic phase was
treated with 2×30 mL of ACN, while the aqueous phase was saturated with
NaCl and subsequently extracted with 2×30 mL of ethyl acetate. The
collected ethyl acetate, ethanol, and ACN were mixed together and dried
[
[
10]
11]
2
o
a) K. Chreptowicz, M. K. Sternicka, P. D. Kowalska, J.
Mierzejewska, Lett. Appl. Microbiol. 2018, 66, 153-160; b) K.
Chreptowicz, M. Wielechowska, J. G. Zubek, E. Rybak, J.
Mierzejewska, Food Bioprod. Process. 2016, 100, 275-281.
D. Vollbrecht, F. Radler, Arch. Microbiol. 1973, 94, 351-358.
a) J.-Y. Hwang, J. Park, J.-H. Seo, M. Cha, B.-K. Cho, J. Kim, B.-G.
Kim, Biotechnol. Bioeng. 2008, 102, 1323-1329; b) J. Liu, J. Jiang,
Y. Bai, T.-p. Fan, Y. Zhao, X. Zheng, Y. Cai, J. Agric. Food Chem.
[
[
12]
13]
2018, 66, 3498-3504; c) Y. Achmon, Z. Ben-Barak Zelas, A.
2 4
with anhydrous Na SO for 12 h. After filtration, the solvents were then
Fishman, Appl. Microbiol. Biotechnol. 2014, 98, 3603-3611.
a) S. Panke, B. Witholt, A. Schmid, M. G. Wubbolts, Appl. Environ.
Microbiol. 1998, 64, 2032-2043; b) S. Wu, Y. Chen, Y. Xu, A. Li, Q.
Xu, A. Glieder, Z. Li, ACS Catal. 2014, 4, 409-420.
D. M. Tieman, H. M. Loucas, J. Y. Kim, D. G. Clark, H. J. Klee, J.
Phytochemistry 2007, 68, 2660-2669.
S. Wu, J. Liu, Z. Li, ACS Catal. 2017, 7, 5225-5233.
F. C. Cochrane, L. B. Davin, N. G. Lewis, J. Phytochemistry 2004,
65, 1557-1564.
evaporated by using a rotary evaporator under reduced pressure to give
the crude product, which was subsequently purified by flash
chromatography on a silica gel column. Ethyl acetate : n-hexane of 1:5
[
14]
[
[
15]
16]
f
was used as the eluent (R ≈ 0.3). 2-PE 6 was obtained in colorless oil (131
mg, 72% yield) and its purity was determined as ≥98% by using GC
analysis. The structure was characterized by 1H-NMR and 13C-NMR. 1H-
[17]
NMR (CDCl
3
, 400 MHz)30 : δ = 7.34-7.23 (5H), 3.87-3.84 (2H), 2.89-2.86
2H) ppm. 13C-NMR (CDCl , 400 MHz)30 : δ = 138.5, 129.1, 128.6, 126.5,
[18]
K. A. P. Payne, M. D. White, K. Fisher, B. Khara, S. S. Bailey, D.
Parker, N. J. W. Rattray, D. K. Trivedi, R. Goodacre, R. Beveridge,
P. Barran, S. E. J. Rigby, N. S. Scrutton, S. Hay, D. Leys, Nature
(
3
63.7, 39.2 ppm
2
015, 522, 497-514.
a) Y. Zhou, S. Wu, Z. Li, Angew. Chem., Int. Ed. 2016, 55, 11647-
1650; b) Y. Zhou, S. Wu, Z. Li, Adv. Synth. Catal. 2017, 359, 4305-
[
19]
20]
1
Acknowledgements
4316.
[
T. Vogl, T. Kickenweiz, J. Pitzer, L. Sturmberger, A. Weninger, B.
W. Biggs, E.-M. Köhler, A. Baumschlager, J. E. Fischer, P. Hyden,
M. Wagner, M. Baumann, N. Borth, M. Geier, P. K. Ajikumar, A.
Glieder, Nature Communications 2018, 9, 3589-3601.
M. Jeschek, D. Gerngross, S. Panke, Nature Communications
2016, 7, 11163-11172.
a) D. J. O'Sullivan, T. R. Klaenhammer, Gene 1993, 137, 227-231;
b) E. Leonard, J. Chemler, K. H. Lim, M. A. G. Koffas, Appl.
Microbiol. Biotechnol. 2006, 70, 85-91; c) D. K. Summers, D. J.
Sherratt, Cell 1984, 36, 1097-1103.
This research was financially supported by Ministry of Education
of Singapore through AcRF Tier 1 Grant (Project No. 279-000-
[
[
21]
22]
477-112).
Keywords: enzyme catalysis • cascade reactions •
biotransformation • L-phenylalanine • 2-phenylethanol • aroma
compounds
[
23]
a) G. Lester, J. Bacteriol 1965, 90, 29-37; b) R. Seward, C. Willetts
Jennifer, M. G. Dinsdale, D. Lloyd, J. Inst. Brew. 2013, 102, 439-
443; c) J. J. Lucchini, J. Corre, A. Cremieux, Res. Microbiol. 1990,
141, 499-510.
[
1]
a) U. T. Bornscheuer, G. W. Huisman, R. J. Kazlauskas, S. Lutz, J.
C. Moore, K. Robins, Nature 2012, 485, 185-194; b) M. T. Reetz, J.
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