Baeyer-Villiger Oxidation of Bridged endo-Tricyclic Ketones with Engineered Escherichia coli Expressing Monooxygenases of Bacterial Origin

Abstract

Whole-cell biotransformations using engineered strains of Escherichia coli expressing cyclopentanone (CPMO) and cyclohexanone monooxygenases (CHMO) of various bacterial origins have been tested for substrate acceptance on tricyclic ketones. Based on the stereopreference of the biocatalytic Baeyer-Villiger oxidation, our recent clustering of this library of enzymes into two distinct groups based on protein sequence was confirmed. Together with short and facile reaction sequences for the production of the substrate ketones, microbial biooxidation enables access to antipodal product lactones as versatile building blocks in natural product and bioactive compound synthesis.

biocatalysis - biooxidation - recombinant whole-cell biotransformation - Baeyer-Villiger oxidation - monooxygenase - stereoselectivity

References

1a Mihovilovic MD. Rudroff F. Grötzl B. Curr. Org. Chem.  2004,  8:  1057 

1b Brink G.-J. Arends IWCE. Sheldon RA. Chem. Rev.  2004,  104:  4105 

2a Bolm C. In Peroxide Chemistry   Adam W. Wiley-VCH; Weinheim: 2000.  p.494-510  

2b Strukul G. Angew. Chem. Int. Ed.  1998,  37:  1198 

3a Kamerbeek NM. Janssen DB. van Berkel WJH. Fraaije MW. Adv. Synth. Catal.  2003,  345:  667 

3b Mihovilovic MD. Müller B. Stanetty P. Eur. J. Org. Chem.  2002,  3711 

3c Stewart JD. Curr. Org. Chem.  1998,  2:  195 

3d Roberts SM. Wan PWH. J. Mol. Catal. B: Enzym.  1998,  4:  111 

3e Willetts A. Trends Biotechnol.  1997,  15:  55 

3f Walsh CT. Chen Y.-CJ. Angew. Chem.  1988,  100:  342 

4 Mihovilovic MD. Rudroff F. Grötzl B. Kapitan P. Snajdrova R. Rydz J. Mach R. Angew. Chem. Int. Ed.  2005,  44:  3609 

5 For another evaluation of some BVMOs of this library see: Kyte BG. Rouviere P. Cheng Q. Stewart JD. J. Org. Chem.  2004,  69:  12 

6a Mihovilovic MD. Rudroff F. Müller B. Stanetty P. Bioorg. Med. Chem. Lett.  2003,  13:  1479 

6b Wang S. Kayser MM. Iwaki H. Lau PCK. J. Mol. Catal. B: Enzym.  2003,  22:  211 

6c Mihovilovic MD. Müller B. Kayser MM. Stanetty P. Synlett  2002,  700 

7a Mihovilovic MD. Kapitan P. Tetrahedron Lett.  2004,  45:  2751 

7b Kelly DR. Knowles CJ. Mahdi JG. Taylor IN. Wright MA. J. Chem. Soc., Chem. Commun.  1995,  729 

7c Alphand V. Furstoss R. J. Org. Chem.  1992,  57:  1306 

8 Stewart JD. Curr. Opin. Biotechnol.  2000,  363 

9 Van Beilen JB. Duetz WA. Schmid A. Witholt B. Trends Biotechnol.  2003,  21:  170 

10 For a S. cerevisiae based whole-cell expression system of BVMOs see: Kayser M. Chen G. Stewart J. Synlett  1999,  153 ; and references therein

11 For an excellent review in biocatalytic desymmetrization reactions see: Garcia-Urdiales E. Alfonso I. Gotor V. Chem. Rev.  2005,  105:  313 

12 Gassman PG. Marshall JL. Org. Synth.  1968,  48:  68 

13 Setzer WN. Whitaker KW. Thompson MA. Yang X.-J. Brown ML. J. Org. Chem.  1992,  57:  2812 

14 Fujita E. Inoue T. Nagao Y. Tetrahedron  1984,  40:  1215 

15a Taschner MJ. Peddada L. J. Chem. Soc., Chem. Commun.  1992,  1384 

15b Taschner MJ. Black DJ. J. Am. Chem. Soc.  1988,  110:  6892 

16 Donoghue NA. Norris DB. Trudgill PW. Eur. J. Biochem.  1976,  63:  175 

17 Brzostowicz P. Walters DM. Thomas SM. Nagarajan V. Rouviere PE. Appl. Environ. Microbiol.  2003,  69:  334 

18 Bramucci MG, Brzostowicz PC, Kostichka KN, Nagarajan V, Rouviere PE, and Thomas SM. inventors; PCT Int. Appl., WO  2003020890.  ; Chem. Abstr. 2003, 138, 233997

19 Brzostowicz PC. Gibson KL. Thomas SM. Blasko MS. Rouviere PE. J. Bacteriol.  2000,  182:  4241 

20 Griffin M. Trudgill PW. Eur. J. Biochem.  1976,  63:  199 

21

endo -Tricyclo[6.2.1.0 ^²,7 ]undecan-11-one ( 1a): yellow oil. ¹H NMR (200 MHz, CDCl₃): δ = 0.81-1.89 (m, 14 H), 2.06-2.14 (m, 2 H). ¹³C NMR (50 MHz, CDCl₃): δ = 17.1 (t), 18.1 (t), 19.5 (t), 32.9 (d), 43.4 (d), 216.7 (s).
endo -Tricyclo[5.2.1.0 ^²,6 ]decan-10-one ( 1b): yellow oil. ¹H NMR (200 MHz, CDCl₃): δ = 1.63-1.75 (m, 12 H), 2.46 (m, 2 H). ¹³C NMR (50 MHz, CDCl₃): δ = 27.7 (t), 29.2 (t), 29.9 (t), 38.2 (d), 43.6 (d), 214.9 (s).
endo -Tricyclo[5.2.1.0 ^²,6 ]dec-8-en-4-one ( 1c): beige amorphous solid; mp = 97-99 °C. ¹H NMR (200 MHz, CDCl₃): δ = 1.30-1.55 (m, 2 H), 1.70-1.90 (m, 2 H), 2.05-2.35 (m, 2 H), 2.70-2.95 (m, 4 H), 6.00-6.15 (m, 2 H). ¹³C NMR (50 MHz, CDCl₃): δ = 39.7 (d), 41.0 (t), 47.0 (d), 49.7 (t), 136.1 (d), 219.9 (s).
endo -Tricyclo[5.2.1.0 ^²,6 ]decan-4-one ( 1d): colorless amorphous solid; mp = 97-100 °C. ¹H NMR (200 MHz, CDCl₃): δ = 1.16-1.59 (m, 6 H), 2.03-2.40 (m, 6 H), 2.51-2.74 (m, 2 H). ¹³C NMR (50 MHz, CDCl₃): δ = 22.1 (t), 38.6 (d), 39.3 (t), 40.6 (t), 41.3 (d), 221.0 (s).

22a Mihovilovic MD. Rudroff F. Grötzl B. Stanetty P. Eur. J. Org. Chem.  2005,  809 

22b Mihovilovic MD. Müller B. Schulze A. Stanetty P. Kayser MM. Eur. J. Org. Chem.  2003,  2243 

23

Typical Procedure for Screening Experiment in Multi-Well Dishes (12 or 24 Wells).
Each well was charged with LB-amp medium (2 mL/12-well format or 1 mL/24-well format) and inoculated with 1% of an overnight preculture of recombinant E. coli strains. A plate was incubated at 120 rpm at 37 °C on an orbital shaker for 2 h. IPTG was added (final concentration of 0.025 mM) together with substrate (1 mg or 0.5 mg) and β-cyclodextrin (1 equiv). The plate was shaken at r.t. for 24 h and then analyzed by chiral phase GC after extraction of the sample with EtOAc.

24a Doig SD. Pickering SCR. Lye GJ. Woodley JM. Biotechnol. Bioeng.  2002,  42 

24b Lye GJ. Dalby PA. Woodley JM. Org. Process Res. Dev.  2002,  6:  434 

25

Physical and Spectroscopic Data of Lactones 2a-d.
endo -9-Oxatricyclo[6.2.2.0 ^²,7 ]dodecan-10-one ( 2a): beige crystals, mp 76-78 °C. ¹H NMR (200 MHz, CDCl₃): δ = 1.09-2.06 (m, 14 H), 2.39-2.41 (q, J = 2.7 Hz, 1 H), 4.36-4.41 (q, J = 3.3 Hz, 2 H). ¹³C NMR (50 MHz, CDCl₃): δ = 16.9 (t), 19.3 (t), 19.5 (t), 20.0 (t), 20.5 (t), 20.8 (t), 32.3 (d), 36.2 (d), 39.8 (d), 78.9 (d), 177.3 (s). Specific optical rotation (CHMO _Brachy ): [α]_D ²⁰ -31.8 (c 0.95, CHCl₃); 99% ee.
endo -8-Oxatricyclo[5.2.2.0 ^²,6 ]undecan-9-one ( 2b): colorless crystals, mp 78-80 °C. ¹H NMR (200 MHz, CDCl₃): δ = 1.66-1.90 (m, 12 H), 2.55 (q, J = 3.1 Hz, 1 H), 4.55 (q, J = 3.7 Hz, 1 H). ¹³C NMR (50 MHz, CDCl₃): δ = 16.7 (t), 20.6 (t), 27.7 (t), 27.9 (t), 28.1 (t), 37.7 (d), 39.2 (d), 41.2 (d), 78.9 (d), 177.3 (s). Specific optical rotation (CHMO _Rhodo2 ): [α]_D ²⁰ -17.3 (c 1.70, CHCl₃); 99% ee.
endo -5-Oxatricyclo[6.2.1.0 ^²,7 ]undec-9-en-4-one ( 2c): colorless oil. ¹H NMR (200 MHz, CDCl₃): δ = 1.45 (d, J = 8.0 Hz, 1 H), 1.64 (d, J = 8.0 Hz, 1 H), 1.93-2.98 (m, 6 H), 3.70 (m, 1 H), 4.30 (m, 1 H), 6.05-6.28 (m, 2 H). ¹³C NMR (50 MHz, CDCl₃): δ = 33.4 (t), 35.9 (d), 38.6 (d), 44.2 (d), 46.0 (d), 50.6 (t), 69.9 (t), 134.9 (d), 136.3 (d), 173.7 (s). Specific optical rotation (CHMO _Brevi2 ): [α]_D ²⁰ -12.7 (c 1.72, CHCl₃); 74% ee.
endo -5-Oxatricyclo[6.2.1.0 ^²,7 ]undecan-4-one ( 2d): colorless solid, mp 78-80 °C. ¹H NMR (200 MHz, CDCl₃): δ = 1.30-1.59 (m, 6 H), 2.12-2.55 (m, 6 H), 4.02-4.32 (m, 2 H). ¹³C NMR (50 MHz, CDCl₃): δ = 21.9 (t), 23.3 (t), 30.4 (t), 35.9 (d), 37.0 (d), 39.0 (d), 40.7 (d), 41.2 (t), 68.2 (t), 174.3 (s). Specific optical rotation (CHMO _Rhodo1 ): [α]_D ²⁰ +32.2 (c 3.53, CHCl₃); 95% ee.

26 Yamamoto H. Katsube J. Sugie A. Shimomura H. Tetrahedron Lett.  1976,  45:  4099 

27 Reetz MT. Brunner B. Schneider T. Schulz F. Clouthier CM. Kayser MM. Angew. Chem. Int. Ed.  2004,  43:  4078 

28 Bocola M. Schulz F. Leca F. Vogel A. Fraaije MW. Reetz MT. Adv. Synth. Catal.  2005,  347:  979 

29 Malito E. Alfieri A. Fraaije MW. Mattevi A. Proc. Natl. Acad. Sci. U.S.A.  2004,  101:  13157