Open AccessStudies on the Production of Fungal Peroxidases in Aspergillus niger
Author(s) -
Ana Conesa,
Cees A. M. J. J. van den Hondel,
Peter J. Punt
Publication year - 2000
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.66.7.3016-3023.2000
Subject(s) - aspergillus niger , phanerochaete , manganese peroxidase , peroxidase , biochemistry , aspergillus oryzae , biology , recombinant dna , extracellular , sodium dodecyl sulfate , microbiology and biotechnology , lignin peroxidase , gel electrophoresis , enzyme , chemistry , gene
To get insight into the limiting factors existing for the efficient production of fungal peroxidase in filamentous fungi, the expression of thePhanerochaete chrysosporium lignin peroxidase H8 (lipA ) and manganese peroxidase (MnP) H4 (mnp1 ) genes inAspergillus niger has been studied. For this purpose, a protease-deficientA. niger strain and different expression cassettes have been used. Northern blotting experiments indicated high steady-state mRNA levels for the recombinant genes. Manganese peroxidase was secreted into the culture medium as an active protein. The recombinant protein showed specific activity and a spectrum profile similar to those of the native enzyme, was correctly processed at its N terminus, and had a slightly lower mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Recombinant MnP production could be increased up to 100 mg/liter upon hemoglobin supplementation of the culture medium. Lignin peroxidase was also secreted into the extracellular medium, although the protein was not active, presumably due to incorrect processing of the secreted enzyme. Expression of thelipA andmnp1 genes fused to theA. niger glucoamylase gene did not result in improved production yields.