Enzymatic Cyclization of 22,23-Dihydro-2,3-oxidosqualene into Euph-7-en-3β-ol and Bacchar-12-en-3β-ol by Recombinant β-Amyrin Synthase

Journal of the American Chemical Society

Volumn 126, Issue 11, 2004, Pages 3426-3427

  Abe, Ikuro ^a   Sakano, Yuichi ^a   Tanaka, Hideya ^a   Lou, Weiwei ^a   Noguchi, Hiroshi ^a   Shibuya, Masaaki ^b   Ebizuka, Yutaka ^b  

^a UNIVERSITY OF SHIZUOKA   (Japan)

^b UNIVERSITY OF TOKYO   (Japan)

Author keywords

[No Author keywords available]

Indexed keywords

22,23 DIHYDRO 2,3 OXIDOSQUALENE; BACCHAR 12 EN 3BETA OL; BETA AMYRIN SYNTHASE; EUPH 7 EN 3BETA OL; RECOMBINANT ENZYME; SPERMINE DERIVATIVE; SQUALENE DERIVATIVE; SYNTHETASE; UNCLASSIFIED DRUG;

ARTICLE; CARBON NUCLEAR MAGNETIC RESONANCE; CRYSTAL STRUCTURE; CYCLIZATION; ENZYME ACTIVE SITE; ENZYME ANALYSIS; ENZYME MECHANISM; ENZYME STRUCTURE; MASS SPECTROMETRY; NONHUMAN; PROTON NUCLEAR MAGNETIC RESONANCE; REACTION ANALYSIS; SACCHAROMYCES CEREVISIAE; STRUCTURE ANALYSIS;

CYCLIZATION; INTRAMOLECULAR TRANSFERASES; LANOSTEROL; PEAS; RECOMBINANT PROTEINS; SQUALENE; TRITERPENES;

EID: 1642340664     PISSN: 00027863     EISSN: None     Source Type: Journal    
DOI: 10.1021/ja031955v     Document Type: Article

References (29)

1. Eschenmoser, A.; Ruzicka, L.; Jeger, O.; Arigoni, D. Helv. Chim. Acta 1955, 38, 1890-1904.
2. For recent reviews, see: (a) Abe, I.; Rohmer, M.; Prestwich, G. D. Chem. Rev. 1993, 93, 2189-2206.
3. (b) Wendt, K. U.; Schulz, G. E.; Corey, E. J.; Liu, D. R. Angew. Chem., Int. Ed. 2000, 39, 2812-2833.
4. (c) Segura, M. J. R.; Jackson, B. E.; Matsuda, S. P. T. Nat. Prod. Rep. 2003, 20, 304-317.
5. (a) Rees, H. H.; Britton, G.; Goodwin, T. W. Biochem. J. 1968, 106, 659-665.
6. (b) Barton, D. H. R.; Mellows, G.; Widdowson, D. A.; Wright, J. J. J. Chem. Soc. (C) 1971, 1142-1148.
7. (c) Seo, S.; Tomita, Y.; Tori, K. J. Am. Chem. Soc. 1981, 103, 2075-2080.
8. (d) Seo, S.; Yoshimura, Y.; Uomori, A.; Takeda, K.; Seto, H.; Ebizuka, Y.; Sankawa, U. J. Am. Chem. Soc. 1988, 110, 1740-1745.
9. For enzyme purification, see: (a) Abe, I.; Sankawa, U.; Ebizuka, Y. Chem. Pharm. Bull. 1989, 37, 536-538.
10. (b) Abe, I.; Ebizuka, Y.; Seo, S.; Sankawa, U. FEBS Lett. 1989, 249, 100-104.
11. (c) Abe, I.; Sankawa, U.; Ebizuka, Y. Chem. Pharm. Bull. 1992, 40, 1755-1760.
12. For cloning and expression, see: (a) Kushiro, T.; Shibuya, M.; Ebizuka, Y. Eur. J. Biochem. 1998, 256, 238-244.
13. (b) Morita, M.; Shibuya, M.; Kushiro, T.; Masuda, K.; Ebizuka, Y. Eur. J. Biochem. 2000, 267, 3453-3460.
14. (a) Wendt, K. U.; Poralla, K.; Schulz, G. E. Science 1997, 277, 1811-1815.
15. (b) Wendt, K. U.; Lenhart, A.; Schulz, G. E. J. Mol. Biol. 1999, 286, 175-187.
16. (a) Kushiro, T.; Shibuya, M.; Ebizuka, Y. J. Am. Chem. Soc. 1999, 121, 1208-1216.
17. (b) Kushiro, T.; Shibuya, M.; Masuda, K.; Ebizuka, Y. J. Am. Chem. Soc. 2000, 122, 6816-6824.
18. 3H-labeled substrate was incubated with a crude cell-free extract from germinating pea seeds. Recrystallizations of the enzyme reaction product with cold bacchar-12-en-3β-ol were repeated to constant specific activity.
19. (a) Corey, E. J.; Gross, S. K. J. Am. Chem. Soc. 1967, 89, 4561-4562.
20. (b) Corey, E. J.; Gross, S. K. J. Am. Chem. Soc. 1968, 90, 5045-5046.
21. (a) Abe, I.; Rohmer, M. J. Chem. Soc., Chem. Commun. 1991, 902-903.
22. (b) Abe, I.; Rohmer, M. J. Chem. Soc., Perkin Trans. 1 1994, 783-791.
23. + 428.3990; calcd 428.4018.
24. 3CN; 1.0 mL/min; 40 °C) to give 3.0 mg of 4 and 0.7 mg of 5, along with 10.5 mg of β-amyrin derived from 2,3-oxidosqualene accumulated in the mutant yeast cells. No other cyclization product was obtained in the reaction mixture, which was confirmed by GC-MS analysis.
25. 3).
26. 1H NMR, 20R-Me of euph-7-en-3β-ol, and 20S-Me of its (20S)-epimer, tirucall-7-en-3β-ol, gave slightly different chemical shifts (δ 0.83 and 0.86, respectively).
27. 3).
28. Akihisa, T.; Kimura, Y.; Tamura, T. Phytochemistry 1994, 37, 1413-1415.
29. Tachibana, K.; Tori, M.; Moriyama, Y.; Tsuyuki, T.; Takahashi, T. Bull. Chem. Soc. Jpn. 1977, 50, 1522-1557.