Discovery and molecular and biocatalytic properties of hydroxynitrile lyase from an invasive millipede, Chamberlinius hualienensis

Mohammad Dadashipour, Yuko Ishida, Kazunori Yamamoto, Yasuhisa Asano

Research output: Contribution to journalArticlepeer-review

42 Citations (Scopus)


Hydroxynitrile lyase (HNL) catalyzes the degradation of cyanohydrins and causes the release of hydrogen cyanide (cyanogenesis). HNL can enantioselectively produce cyanohydrins, which are valuable building blocks for the synthesis of fine chemicals and pharmaceuticals, and is used as an important biocatalyst in industrial biotechnology. Currently, HNLs are isolated from plants and bacteria. Because industrial biotechnology requires more efficient and stable enzymes for sustainable development, we must continuously explore other potential enzyme sources for the desired HNLs. Despite the abundance of cyanogenic millipedes in the world, there has been no precise study of the HNLs from these arthropods. Here we report the isolation of HNL from the cyanide-emitting invasive millipede Chamberlinius hualienensis, along with its molecular properties and application in biocatalysis. The purified enzyme displays a very high specific activity in the synthesis of mandelonitrile. It is a glycosylated homodimer protein and shows no apparent sequence identity or homology with proteins in the known databases. It shows biocatalytic activity for the condensation of various aromatic aldehydes with potassium cyanide to produce cyanohydrins and has high stability over a wide range of temperatures and pH values. It catalyzes the synthesis of (R)-mandelonitrile from benzaldehyde with a 99% enantiomeric excess, without using any organic solvents. Arthropod fauna comprise 80% of terrestrial animals. We propose that these animals can be valuable resources for exploring not only HNLs but also diverse, efficient, and stable biocatalysts in industrial biotechnology.

Original languageEnglish
Pages (from-to)10605-10610
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number34
Publication statusPublished - 25 Aug 2015
Externally publishedYes


  • Acetonitriles/metabolism
  • Aldehyde-Lyases/chemistry
  • Aldehydes/metabolism
  • Animals
  • Arthropods/enzymology
  • Base Sequence
  • Benzaldehydes/metabolism
  • Biocatalysis
  • DNA, Complementary/genetics
  • Glycosylation
  • Hydrogen-Ion Concentration
  • Molecular Sequence Data
  • Molecular Structure
  • Nitriles/metabolism
  • Organ Specificity
  • Protein Processing, Post-Translational
  • Protein Structure, Secondary
  • Sequence Homology, Amino Acid
  • Stereoisomerism


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