The development of an immobilized lactate oxidase system for lactic acid analysis.


Journal Article

An enzyme designated as lactate oxidase was purified from Acetobacter peroxydans by using the partition methods of separation. A DE-52 cellulose column was used for the primary purification of lactate oxidase, and the purified enzyme was covalently bound to a porous cellulose bead matrix in which benzoquinone was used as the coupling reagent. The physicochemical properties of the native and immobilized enzymes were determined including molecular weight, cofactor requirements, and optimal reaction conditions. Lactate oxidase was shown not to be subject to product inhibition, and to require Mg(2+) as a metal cofactor. Analysis of an immobilized lactate oxidase packed-bed reactor indicated that this system may not be subject to internal diffusional limitations. Molecular oxygen appeared to be a cosubstrate of the enzyme, and a reaction mechanism was postulated to predict the kinetic behavior of the immobilized reactor system. Applications of the immobilized lactate oxidase reactor for the pulse-flow analysis of lactic acid in whole milk and in a yeast fermentation system were considered.

Full Text

Cited Authors

  • Cannon, JJ; Chen, LF; Flickinger, MC; Tsao, GT

Published Date

  • February 1, 1984

Published In

Volume / Issue

  • 26 / 2

Start / End Page

  • 167 - 173

PubMed ID

  • 18551703

Pubmed Central ID

  • 18551703

Electronic International Standard Serial Number (EISSN)

  • 1097-0290

International Standard Serial Number (ISSN)

  • 0006-3592

Digital Object Identifier (DOI)

  • 10.1002/bit.260260209


  • eng