Skip to Content
Merck
  • The antibrowning agent sulfite inactivates Agaricus bisporus tyrosinase through covalent modification of the copper-B site.

The antibrowning agent sulfite inactivates Agaricus bisporus tyrosinase through covalent modification of the copper-B site.

The FEBS journal (2013-10-12)
Tomas F M Kuijpers, Harry Gruppen, Stefano Sforza, Willem J H van Berkel, Jean-Paul Vincken
ABSTRACT

Sulfite salts are widely used as antibrowning agents in food processing. Nevertheless, the exact mechanism by which sulfite prevents enzymatic browning has remained unknown. Here, we show that sodium hydrogen sulfite (NaHSO3) irreversibly blocks the active site of tyrosinase from the edible mushroom Agaricus bisporus, and that the competitive inhibitors tropolone and kojic acid protect the enzyme from NaHSO3 inactivation. LC-MS analysis of pepsin digests of NaHSO3 -treated tyrosinase revealed two peptides showing a neutral loss corresponding to the mass of SO3 upon MS(2) fragmentation. These peptides were found to be homologous peptides containing two of the three histidine residues that form the copper-B-binding site of mushroom tyrosinase isoform PPO3 and mushroom tyrosinase isoform PPO4, which were both present in the tyrosinase preparation used. Peptides showing this neutral loss behavior were not found in the untreated control. Comparison of the effects of NaHSO3 on apo-tyrosinase and holo-tyrosinase indicated that inactivation is facilitated by the active site copper ions. These data provide compelling evidence that inactivation of mushroom tyrosinase by NaHSO3 occurs through covalent modification of a single amino-acid residue, probably via addition of HSO3(-) to one of the copper-coordinating histidines in the copper-B site of the enzyme.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-Histidine, BioUltra, ≥99.5% (NT)
Sigma-Aldrich
L-Histidine, suitable for cell culture, meets EP, USP testing specifications, from non-animal source
Sigma-Aldrich
L-Histidine, ReagentPlus®, ≥99% (TLC)
Sigma-Aldrich
Tyrosinase from mushroom, lyophilized powder, ≥1000 unit/mg solid
Supelco
L-Histidine, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Supelco
L-Histidine, Pharmaceutical Secondary Standard; Certified Reference Material
SAFC
L-Histidine
Sigma-Aldrich
Tropolone, 98%
Sigma-Aldrich
Kojic acid