Does nature click? Theoretical prediction of an enzyme-catalyzed transannular 1,3-dipolar cycloaddition in the biosynthesis of lycojaponicumins A and B

J Am Chem Soc. 2013 Nov 20;135(46):17638-42. doi: 10.1021/ja409928z. Epub 2013 Nov 6.

Abstract

Biosynthetic 1,3-dipolar cycloadditions are rare. No enzymes have yet been identified whose function is to catalyze this class of reactions. Recently, however, a 1,3-dipolar cycloaddition was proposed as a key step in the biosynthesis of two Lycopodium alkaloids, lycojaponicumins A and B. The lycojaponicumins' fused bicyclic tetrahydroisoxazole ring system was proposed to originate from a transannular 1,3-dipolar cycloaddition between a nitrone and an enone in a nine-membered macrocycle. We have used quantum mechanical calculations to predict whether this cycloaddition could constitute a feasible step in a biosynthetic pathway. Our calculations define a general computational approach for analyzing whether a putative biosynthetic reaction is likely to be enzyme-catalyzed. The quantum mechanically predicted rate of the uncatalyzed reaction in water is compared with the rate enhancement theoretically achievable when the reaction is catalyzed by a theozyme (theoretical enzyme). Density functional theory calculations (M06-2X) predict that the uncatalyzed transannular 1,3-dipolar cycloaddition of the putative lycojaponicumin precursor in water is moderately facile (ΔG(++) = 21.5 kcal/mol, k = 10(-3) s(-1)) and that an enzyme could accelerate the cycloaddition by placing hydrogen bond donors around the enone while maintaining an otherwise nonpolar active site. The theoretical enzyme-catalyzed process has ΔG(++) ≈ 17 kcal/mol, corresponding to a 2000-fold rate enhancement, and the predicted kcat (2 s(-1)) is similar to those of known enzymes involved in secondary metabolic pathways. Thus, theory predicts that the proposed transannular 1,3-dipolar cycloaddition is a plausible step in a biosynthetic pathway leading to the lycojaponicumins and suggests that dipolar cycloadditions can be accelerated by enzyme catalysis.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Alkaloids / biosynthesis*
  • Alkaloids / chemistry
  • Azabicyclo Compounds / chemistry
  • Biocatalysis
  • Cyclization
  • Enzymes / metabolism*
  • Molecular Structure
  • Quantum Theory*
  • Thermodynamics

Substances

  • Alkaloids
  • Azabicyclo Compounds
  • Enzymes
  • lycojaponicumin A
  • lycojaponicumin B