Affiliation:
1. Department of Molecular Genetics and Gene Technology, TNO Nutrition and Food Research Institute, 3704 HE Zeist, The Netherlands
Abstract
ABSTRACT
To get insight into the limiting factors existing for the efficient production of fungal peroxidase in filamentous fungi, the expression of the
Phanerochaete chrysosporium
lignin peroxidase H8 (
lipA
) and manganese peroxidase (MnP) H4 (
mnp1
) genes in
Aspergillus niger
has been studied. For this purpose, a protease-deficient
A. 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 the
lipA
and
mnp1
genes fused to the
A. niger
glucoamylase gene did not result in improved production yields.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
115 articles.
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