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The role of the Aspergillus niger furin-type protease gene in processing of fungal proproteins and fusion proteins: Evidence for alternative processing of recombinant (fusion-) proteins

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Author: Punt, P.J. · Drint-Kuijvenhoven, A. · Lokman, B.C. · Spencer, J.A. · Jeenes, D. · Archer, D.A. · Hondel, C.A.M.J.J. van den
Type:article
Date:2003
Source:Journal of Biotechnology, 1, 106, 23-32
Identifier: 237524
doi: doi:10.1016/j.jbiotec.2003.09.005
Keywords: Biology · Biotechnology · Filamentous fungi · Fusion-proteins · Protease · Protein processing · Protein secretion · Secretion carrier · Biotechnology · Genes · Growth kinetics · Mutagenesis · Proteins · Cleavage sites · Genetic engineering · arginine · aspergillopepsin · fungal protein · furin · glucan 1,4 alpha glucosidase · hybrid protein · lysine · protein precursor · proteinase · recombinant protein · unclassified drug · allele · amino acid composition · article · Aspergillus niger · controlled study · fungal gene · fungal strain · fungus mutant · gene deletion · gene disruption · gene function · genetic code · genetic recombination · molecular cloning · nonhuman · nucleotide sequence · priority journal · protein degradation · protein processing · wild type · Aspergillus niger · Endopeptidases · Fungal Proteins · Furin · Gene Expression Regulation, Enzymologic · Gene Expression Regulation, Fungal · Protein Processing, Post-Translational · Recombinant Fusion Proteins · Trans-Activation (Genetics) · Aspergillus · Aspergillus niger · Fungi

Abstract

We have characterized growth and protein processing characteristics of Aspergillus niger strains carrying a disrupted allele of the previously cloned and characterized kexB gene [Appl. Environ. Microbiol. 66 (2000) 363] encoding a furin-type endoprotease. Deletion of the single-copy gene confirms it to be non-essential but disruptant strains exhibit a morphologically distinct phenotype characterized by hyperbranching. Processing of homologous pro-proteins and fusion proteins comprised of a heterologous protein fused down-stream of glucoamylase and separated at the fusion junction by an endoproteolytic cleavage site was compared in wildtype and mutant strains of A. niger. We show that maturation of the native glucoamylase requires KexB, whereas maturation of aspergillopepsin does not. The processing of fusion proteins carrying Lys-Arg requires KexB, although alternative endoproteases are capable of cleaving protein fusions at sites adjacent to Lys-Arg. © 2003 Elsevier B.V. All rights reserved.