Title
Studies on the production of fungal peroxidases in Aspergillus niger
Author
Conesa, A.
van den Hondel, C.A.M.J.J.
Punt, P.J.
Centraal Instituut voor Voedingsonderzoek TNO
Publication year
2000
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. Chemicals/CAS: Ferrous Compounds; Heme, 14875-96-8; lignin peroxidase, EC 1.11.1.-; manganese peroxidase, EC 1.11.1.13; Peroxidases, EC 1.11.1.-; Recombinant Proteins
Subject
Nutrition
Botany
Fungi
Fungal enzyme
Glucan 1,4 alpha glucosidase
Hemoglobin
Lignin peroxidase
Manganese peroxidase
Messenger RNA
Peroxidase
Recombinant protein
Aspergillus niger
Enzyme activity
Enzyme purification
Enzyme release
Enzyme synthesis
Gene overexpression
Nonhuman
Nucleotide sequence
Protein expression
Protein processing
Aspergillus niger
Blotting, Northern
Blotting, Western
Ferrous Compounds
Heme
Peroxidases
Phanerochaete
Recombinant Proteins
Transformation, Genetic
Aspergillus niger
Chrysosporium
Fungi
Phanerochaete chrysosporium
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DOI
https://doi.org/10.1128/aem.66.7.3016-3023.2000
TNO identifier
56789
ISSN
0099-2240
Source
Applied and Environmental Microbiology, 66 (7), 3016-3023
Document type
article