Characterization of a cryptic, pyrroloquinoline quinone-dependent dehydrogenase of Gluconobacter sp. strain CHM43

Biosci Biotechnol Biochem. 2021 Mar 24;85(4):998-1004. doi: 10.1093/bbb/zbab005.

Abstract

We characterized the pyrroloquinoline quinone (PQQ)-dependent dehydrogenase 9 (PQQ-DH9) of Gluconobacter sp. strain CHM43, which is a homolog of PQQ-dependent glycerol dehydrogenase (GLDH). We used a plasmid construct to express PQQ-DH9. The expression host was a derivative strain of CHM43, which lacked the genes for GLDH and the membrane-bound alcohol dehydrogenase and consequently had minimal ability to oxidize primary and secondary alcohols. The membranes of the transformant exhibited considerable d-arabitol dehydrogenase activity, whereas the reference strain did not, even if it had PQQ-DH9-encoding genes in the chromosome and harbored the empty vector. This suggests that PQQ-DH9 is not expressed in the genome. The activities of the membranes containing PQQ-DH9 and GLDH suggested that similar to GLDH, PQQ-DH9 oxidized a wide variety of secondary alcohols but had higher Michaelis constants than GLDH with regard to linear substrates such as glycerol. Cyclic substrates such as cis-1,2-cyclohexanediol were readily oxidized by PQQ-DH9.

Keywords: PQQ-dependent dehydrogenase; acetic acid bacteria; orphan enzyme; pyrroloquinoline quinone.

MeSH terms

  • Alcohol Dehydrogenase / metabolism
  • Genome, Bacterial
  • Gluconobacter / metabolism*
  • Oxidoreductases / metabolism*
  • PQQ Cofactor / metabolism*
  • Plasmids
  • Sugar Alcohols / metabolism

Substances

  • Sugar Alcohols
  • PQQ Cofactor
  • Oxidoreductases
  • Alcohol Dehydrogenase
  • arabitol