Global to push GA events into
skip to main content

Title: In vivo and in vitro olefin cyclopropanation catalyzed by heme enzymes

The present invention provides methods for catalyzing the conversion of an olefin to any compound containing one or more cyclopropane functional groups using heme enzymes. In certain aspects, the present invention provides a method for producing a cyclopropanation product comprising providing an olefinic substrate, a diazo reagent, and a heme enzyme; and admixing the components in a reaction for a time sufficient to produce a cyclopropanation product. In other aspects, the present invention provides heme enzymes including variants and fragments thereof that are capable of carrying out in vivo and in vitro olefin cyclopropanation reactions. Expression vectors and host cells expressing the heme enzymes are also provided by the present invention.
Inventors:
; ; ; ;
Issue Date:
OSTI Identifier:
1332273
Assignee:
CALIFORNIA INSTITUTE OF TECHNOLOGY (Pasadena, CA) CHO
Patent Number(s):
9,493,799
Application Number:
14/625,449
Contract Number:
FG02-06ER15762
Resource Relation:
Patent File Date: 2015 Feb 18
Research Org:
CALIFORNIA INSTITUTE OF TECHNOLOGY, Pasadena, CA (United States)
Sponsoring Org:
USDOE
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY

Other works cited in this record:

PHENIX: a comprehensive Python-based system for macromolecular structure solution
journal, January 2010
  • Adams, Paul D.; Afonine, Pavel V.; Bunkóczi, Gábor
  • Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2, p. 213-221
  • DOI: 10.1107/S0907444909052925

Basic local alignment search tool
journal, October 1990
  • Altschul, Stephen F.; Gish, Warren; Miller, Webb
  • Journal of Molecular Biology, Vol. 215, Issue 3, p. 403-410
  • DOI: 10.1016/S0022-2836(05)80360-2

The CCP4 suite programs for protein crystallography
journal, September 1994

Organometallic alkane CH activation
journal, November 2004

Protein stability promotes evolvability
journal, March 2006
  • Bloom, J. D.; Labthavikul, S. T.; Otey, C. R.
  • Proceedings of the National Academy of Sciences, Vol. 103, Issue 15, p. 5869-5874
  • DOI: 10.1073/pnas.0510098103

Catalytic Promiscuity in Biocatalysis: Using Old Enzymes to Form New Bonds and Follow New Pathways
journal, November 2004
  • Bornscheuer, Uwe T.; Kazlauskas, Romas J.
  • Angewandte Chemie International Edition, Vol. 43, Issue 45, p. 6032-6040
  • DOI: 10.1002/anie.200460416

The role of Thr268 and Phe393 in cytochrome P450 BM3
journal, May 2006
  • Clark, Jonathan P.; Miles, Caroline S.; Mowat, Christopher G.
  • Journal of Inorganic Biochemistry, Vol. 100, Issue 5-6, p. 1075-1090
  • DOI: 10.1016/j.jinorgbio.2005.11.020

Synthesis of cyclopropane containing natural products
journal, October 2001

Probing the molecular determinants of coenzyme selectivity in the P450 BM3 FAD/NADPH domain
journal, August 2009
  • Dunford, Adrian J.; Girvan, Hazel M.; Scrutton, Nigel S.
  • Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics, Vol. 1794, Issue 8, p. 1181-1189
  • DOI: 10.1016/j.bbapap.2009.03.014

Coot model-building tools for molecular graphics
journal, November 2004
  • Emsley, Paul; Cowtan, Kevin
  • Acta Crystallographica Section D Biological Crystallography, Vol. 60, Issue 12, p. 2126-2132
  • DOI: 10.1107/S0907444904019158

Bis(oxazolines) as chiral ligands in metal-catalyzed asymmetric reactions. Catalytic, asymmetric cyclopropanation of olefins
journal, January 1991
  • Evans, David A.; Woerpel, Keith A.; Hinman, Mira M.
  • Journal of the American Chemical Society, Vol. 113, Issue 2, p. 726-728
  • DOI: 10.1021/ja00002a080

Scaling and assessment of data quality
journal, December 2005
  • Evans, Philip
  • Acta Crystallographica Section D Biological Crystallography, Vol. 62, Issue 1, p. 72-82
  • DOI: 10.1107/S0907444905036693

C-H bond activation in heme proteins: the role of thiolate ligation in cytochrome P450
journal, February 2009

The bioinorganic chemistry of iron in oxygenases and supramolecular assemblies
journal, March 2003
  • Groves, J. T.
  • Proceedings of the National Academy of Sciences, Vol. 100, Issue 7, p. 3569-3574
  • DOI: 10.1073/pnas.0830019100

General Method for Sequence-independent Site-directed Chimeragenesis
journal, July 2003

Complex reactions catalyzed by cytochrome P450 enzymes
journal, March 2007
  • Isin, Emre M.; Guengerich, F. Peter
  • Biochimica et Biophysica Acta (BBA) - General Subjects, Vol. 1770, Issue 3, p. 314-329
  • DOI: 10.1016/j.bbagen.2006.07.003

Integration, scaling, space-group assignment and post-refinement
journal, January 2010
  • Kabsch, Wolfgang
  • Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2, p. 133-144
  • DOI: 10.1107/S0907444909047374

Chemoenzymatic elaboration of monosaccharides using engineered cytochrome P450BM3 demethylases
journal, September 2009
  • Lewis, J. C.; Bastian, S.; Bennett, C. S.
  • Proceedings of the National Academy of Sciences, Vol. 106, Issue 39, p. 16550-16555
  • DOI: 10.1073/pnas.0908954106

Combinatorial Alanine Substitution Enables Rapid Optimization of Cytochrome P450BM3 for Selective Hydroxylation of Large Substrates
journal, November 2010
  • Lewis, Jared C.; Mantovani, Simone M.; Fu, Yu
  • ChemBioChem, Vol. 11, Issue 18, p. 2502-2505
  • DOI: 10.1002/cbic.201000565

Engineering Cytochrome P450 BM3 for Terminal Alkane Hydroxylation
journal, April 2006
  • Meinhold, Peter; Peters, Matthew W.; Hartwick, Adam
  • Advanced Synthesis & Catalysis, Vol. 348, Issue 6, p. 763-772
  • DOI: 10.1002/adsc.200505465

Recent developments in asymmetric cyclopropanation
journal, July 2008

Molecular basis for the inability of an oxygen atom donor ligand to replace the natural sulfur donor heme axial ligand in cytochrome P450 catalysis. Spectroscopic characterization of the Cys436Ser CYP2B4 mutant
journal, March 2011
  • Perera, Roshan; Sono, Masanori; Voegtle, Heather L.
  • Archives of Biochemistry and Biophysics, Vol. 507, Issue 1, p. 119-125
  • DOI: 10.1016/j.abb.2010.12.010

Semisynthesis of axial-ligand (position 80) mutants of cytochrome c
journal, January 1991
  • Raphael, Adrienne L.; Gray, Harry B.
  • Journal of the American Chemical Society, Vol. 113, Issue 3, p. 1038-1040
  • DOI: 10.1021/ja00003a045

Catalyst structure and the enantioselective cyclopropanation of alkenes by copper complexes of biaryldiimines: the importance of ligand acceleration
journal, May 2001
  • Sanders, Christopher J.; Gillespie, Kevin M.; Scott, Peter
  • Tetrahedron: Asymmetry, Vol. 12, Issue 7, p. 1055-1061
  • DOI: 10.1016/S0957-4166(01)00157-4

MOLREP an Automated Program for Molecular Replacement
journal, December 1997

Replacement of active-site cysteine-436 by serine converts cytochrome P450 2B4 into an NADPH oxidase with negligible monooxygenase activity
journal, September 2002
  • Vatsis, Kostas P.; Peng, Hwei-Ming; Coon, Minor J.
  • Journal of Inorganic Biochemistry, Vol. 91, Issue 4, p. 542-553
  • DOI: 10.1016/S0162-0134(02)00438-5

Production of amorphadiene in yeast, and its conversion to dihydroartemisinic acid, precursor to the antimalarial agent artemisinin
journal, January 2012
  • Westfall, P. J.; Pitera, D. J.; Lenihan, J. R.
  • Proceedings of the National Academy of Sciences, Vol. 109, Issue 3, p. E111-E118
  • DOI: 10.1073/pnas.1110740109

Protein engineering as a tool for understanding electron transfer
journal, August 1993

BuMgNiPr2: A New Base for Stoichiometric, Position-Selective Deprotonation of Cyclopropane Carboxamides and Other Weak CH Acids
journal, June 2002

Similar records in DOepatents and OSTI.GOV collections: