title: Calnexin overexpression increases manganese peroxidase production in Aspergillus niger.
abstract: Heme-containing peroxidases from white rot basidiomycetes, in contrast to most proteins of fungal origin, are poorly produced in industrial filamentous fungal strains. Factors limiting peroxidase production are believed to operate at the posttranslational level. In particular, insufficient availability of the prosthetic group which is required for peroxidase biosynthesis has been proposed to be an important bottleneck. In this work, we analyzed the role of two components of the secretion pathway, the chaperones calnexin and binding protein (BiP), in the production of a fungal peroxidase. Expression of the Phanerochaete chrysosporium manganese peroxidase (MnP) in Aspergillus niger resulted in an increase in the expression level of the clxA and bipA genes. In a heme-supplemented medium, where MnP was shown to be overproduced to higher levels, induction of clxA and bipA was also higher. Overexpression of these two chaperones in an MnP-producing strain was analyzed for its effect on MnP production. Whereas bipA overexpression seriously reduced MnP production, overexpression of calnexin resulted in a four- to fivefold increase in the extracellular MnP levels. However, when additional heme was provided in the culture medium, calnexin overexpression had no synergistic effect on MnP production. The possible function of these two chaperones in MnP maturation and production is discussed.
authors.affiliation: Department of Applied Microbiology and Gene Technology, TNO Food and Nutrition Research Institute, 3700 AJ Zeist, The Netherlands.
authors.forename: Ana
authors.initials: A
authors.lastname: Conesa
authors.forename: David
authors.initials: D
authors.lastname: Jeenes
authors.forename: David B
authors.initials: DB
authors.lastname: Archer
authors.forename: Cees A M J J
authors.initials: CA
authors.lastname: van den Hondel
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Punt
mesh_terms: D001234:Aspergillus niger / Q000201:enzymology* / Q000235:genetics; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D037281:Calnexin; D003470:Culture Media; D005656:Fungal Proteins; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D006418:Heme / Q000378:metabolism; D010544:Peroxidases / Q000096:biosynthesis* / Q000235:genetics; D020075:Phanerochaete / Q000201:enzymology / Q000235:genetics; D014170:Transformation, Genetic
chemical_list: D002135:Calcium-Binding Proteins; D003470:Culture Media; D005656:Fungal Proteins; D018840:HSP70 Heat-Shock Proteins; D037281:Calnexin; D006418:Heme; D010544:Peroxidases; C051129:manganese peroxidase