Enzyme-catalysed [4+2] cycloaddition is a key step in the biosynthesis of spinosyn A

Hak Joong Kim, Mark W. Ruszczycky, Sei Hyun Choi, Yung Nan Liu, Hung Wen Liu

Research output: Contribution to journalArticle

171 Citations (Scopus)

Abstract

The Diels-Alder reaction is a [4+2] cycloaddition reaction in which a cyclohexene ring is formed between a 1,3-diene and an electron-deficient alkene via a single pericyclic transition state. This reaction has been proposed as a key transformation in the biosynthesis of many cyclohexene-containing secondary metabolites. However, only four purified enzymes have thus far been implicated in biotransformations that are consistent with a Diels-Alder reaction, namely solanapyrone synthase, LovB, macrophomate synthase, and riboflavin synthase. Although the stereochemical outcomes of these reactions indicate that the product formation could be enzyme-guided in each case, these enzymes typically demonstrate more than one catalytic activity, leaving their specific influence on the cycloaddition step uncertain. In our studies of the biosynthesis of spinosyn A, a tetracyclic polyketide-derived insecticide from Saccharopolyspora spinosa, we identified a cyclase, SpnF, that catalyses a transannular [4+2] cycloaddition to form the cyclohexene ring in spinosyn A. Kinetic analysis demonstrates that SpnF specifically accelerates the ring formation reaction with an estimated 500-fold rate enhancement. A second enzyme, SpnL, was also identified as responsible for the final cross-bridging step that completes the tetracyclic core of spinosyn A in a manner consistent with a Rauhut-Currier reaction. This work is significant because SpnF represents the first example characterized in vitro of a stand-alone enzyme solely committed to the catalysis of a [4+2] cycloaddition reaction. In addition, the mode of formation of the complex perhydro-as-indacene moiety in spinosyn A is now fully established.

Original languageEnglish
Pages (from-to)109-112
Number of pages4
JournalNature
Volume473
Issue number7345
DOIs
Publication statusPublished - 2011 May 5
Externally publishedYes

Fingerprint

Cycloaddition Reaction
Enzymes
Riboflavin Synthase
Saccharopolyspora
Polyketides
Alkenes
Insecticides
Biotransformation
Catalysis
spinosyn A
Electrons
cyclohexene

ASJC Scopus subject areas

  • General

Cite this

Kim, H. J., Ruszczycky, M. W., Choi, S. H., Liu, Y. N., & Liu, H. W. (2011). Enzyme-catalysed [4+2] cycloaddition is a key step in the biosynthesis of spinosyn A. Nature, 473(7345), 109-112. https://doi.org/10.1038/nature09981

Enzyme-catalysed [4+2] cycloaddition is a key step in the biosynthesis of spinosyn A. / Kim, Hak Joong; Ruszczycky, Mark W.; Choi, Sei Hyun; Liu, Yung Nan; Liu, Hung Wen.

In: Nature, Vol. 473, No. 7345, 05.05.2011, p. 109-112.

Research output: Contribution to journalArticle

Kim, HJ, Ruszczycky, MW, Choi, SH, Liu, YN & Liu, HW 2011, 'Enzyme-catalysed [4+2] cycloaddition is a key step in the biosynthesis of spinosyn A', Nature, vol. 473, no. 7345, pp. 109-112. https://doi.org/10.1038/nature09981
Kim, Hak Joong ; Ruszczycky, Mark W. ; Choi, Sei Hyun ; Liu, Yung Nan ; Liu, Hung Wen. / Enzyme-catalysed [4+2] cycloaddition is a key step in the biosynthesis of spinosyn A. In: Nature. 2011 ; Vol. 473, No. 7345. pp. 109-112.
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