|Title||Expression of the Caldariomyces fumago chloroperoxidase in Aspergillus niger and characterization of the recombinant enzyme|
|Publication Type||Journal Article|
|Year of Publication||2001|
|Authors||Conesa, A, van De Velde, F, van Rantwijk, F, Sheldon, RA, van den Hondel, CA, Punt, PJ|
|Journal||J Biol Chem|
|Keywords||Aspergillus niger/enzymology/genetics Catalysis Chloride Peroxidase/biosynthesis/*genetics Fungal Proteins/biosynthesis/*genetics Recombinant Proteins/biosynthesis/genetics Substrate Specificity|
The Caldariomyces fumago chloroperoxidase was successfully expressed in Aspergillus niger. The recombinant enzyme was produced in the culture medium as an active protein and could be purified by a three-step purification procedure. The catalytic behavior of recombinant chloroperoxidase (rCPO) was studied and compared with that of native CPO. The specific chlorination activity (47 units/nmol) of rCPO and its pH optimum (pH 2.75) were very similar to those of native CPO. rCPO catalyzes the oxidation of various substrates in comparable yields and selectivities to native CPO. Indole was oxidized to 2-oxindole with 99% selectivity and thioanisole to the corresponding R-sulfoxide (enantiomeric excess >98%). Incorporation of (18)O from labeled H(2)18O(2) into the oxidized products was 100% in both cases.
Conesa, A van De Velde, F van Rantwijk, F Sheldon, R A van Den Hondel, C A Punt, P J Research Support, Non-U.S. Gov’t United States The Journal of biological chemistry J Biol Chem. 2001 May 25;276(21):17635-40. Epub 2001 Feb 22.