A simple assay for 6-aminohexanoate-oligomer-hydrolase using N-(4-nitrophenyl)-6-aminohexanamide

J Biosci Bioeng. 2012 Dec;114(6):586-8. doi: 10.1016/j.jbiosc.2012.07.006. Epub 2012 Aug 4.

Abstract

We synthesized N-(4-nitrophenyl)-6-aminohexanamide (AHpNA) and used it as a substrate in a kinetic study of 6-aminohexanoate-hydrolase (NylB), a nylon oligomer-hydrolyzing enzyme. NylBs derived from Arthrobacter sp. KI72 and Pseudomonas sp. NK87 hydrolyzed AHpNA as well as a 6-aminohexanoic acid dimer, a known substrate for NylB. The K(m) values of the NylB from Arthrobacter sp. KI72 and Pseudomonas sp. NK87 for AHpNA were 0.5 mM and 2.0 mM, respectively.

MeSH terms

  • Amidohydrolases / analysis
  • Amidohydrolases / metabolism*
  • Aminocaproates
  • Aminocaproic Acid / chemistry
  • Aminocaproic Acid / metabolism
  • Arthrobacter / enzymology
  • Enzyme Assays / methods*
  • Hydrolysis
  • Kinetics
  • Nitrophenols / chemical synthesis
  • Nitrophenols / chemistry
  • Nitrophenols / metabolism*
  • Pseudomonas / enzymology
  • Substrate Specificity

Substances

  • 6-aminohexanoic acid cyclic dimer
  • Aminocaproates
  • N-(4-nitrophenyl)-6-aminohexanamide
  • Nitrophenols
  • Amidohydrolases
  • 6-aminohexanoate-dimer hydrolase
  • Aminocaproic Acid