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Oxidation of Alkenes: Syn Dihydroxylation with Osmium Tetraoxide02:44

Oxidation of Alkenes: Syn Dihydroxylation with Osmium Tetraoxide

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Introduction
One of the convenient methods for the preparation of aldehydes and ketones is via hydration of alkynes. Hydroboration-oxidation of alkynes is an indirect hydration reaction in which an alkyne is treated with borane followed by oxidation with alkaline peroxide to form an enol that rapidly converts into an aldehyde or a ketone. Terminal alkynes form aldehydes, whereas internal alkynes give ketones as the final product.
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Epoxides that are three-membered ring systems are more reactive than other cyclic and acyclic ethers. The high reactivity of epoxides originates from the strain present in the ring. This ring strain acts as a driving force for epoxides to undergo ring-opening reactions either with halogen acids or weak nucleophiles in the presence of mild acid. The acid catalyst converts the epoxide oxygen, a poor leaving group, into an oxonium ion, a better leaving group, making the reaction feasible. The...
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New bioorganic reagents: evolved cyclohexanone monooxygenase--why is it more selective?

Margaret M Kayser1, Christopher M Clouthier

  • 1Department of Physical Sciences, University of New Brunswick, Saint John, New Brunswick, E2L 4L5 Canada. kayser@unbsj.ca

The Journal of Organic Chemistry
|October 27, 2006
PubMed
Summary

Researchers engineered cyclohexanone monooxygenase (CHMO) mutants for Baeyer-Villiger oxidation. A single mutation (Phe432Ser) yielded a robust and selective catalyst, outperforming the wild-type enzyme for various substrates.

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Area of Science:

  • Biocatalysis and Enzyme Engineering
  • Organic Chemistry
  • Stereoselective Synthesis

Background:

  • Cyclohexanone monooxygenase (CHMO) is a key enzyme for Baeyer-Villiger oxidation.
  • Enzyme evolution has generated CHMO variants with altered catalytic properties.
  • Understanding structure-activity relationships is crucial for optimizing biocatalysts.

Purpose of the Study:

  • To evaluate four evolved CHMO mutants as catalysts for Baeyer-Villiger oxidation of diverse cyclohexanone derivatives.
  • To identify optimal mutant/substrate combinations for efficient biotransformations.
  • To investigate the impact of specific mutations on catalyst robustness and selectivity.

Main Methods:

  • Directed evolution of cyclohexanone monooxygenase (CHMO).
  • Baeyer-Villiger oxidation of 4-hydroxycyclohexanone and its derivatives.
  • Laboratory-scale biotransformations with full product characterization.
  • Determination of absolute configurations for selected products.

Main Results:

  • Several excellent catalytic matches between CHMO mutants and various 4-substituted/4,4-disubstituted cyclohexanones were identified.
  • A single-point mutant (Phe432Ser) demonstrated comparable robustness and superior selectivity to wild-type CHMO in several cases.
  • Product characterization confirmed the efficiency and specificity of the engineered biocatalysts.

Conclusions:

  • Engineered CHMO mutants offer improved catalytic performance for Baeyer-Villiger oxidations.
  • The Phe432Ser mutation represents a significant advancement in CHMO-based biocatalysis, enhancing selectivity.
  • A proposed model highlights the role of the 432 serine residue in controlling enantioselectivity, paving the way for further enzyme design.