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

article






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
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.
Topics























































BiotechnologyFilamentous fungiFusion-proteinsProteaseProtein processingProtein secretionSecretion carrierBiotechnologyGenesGrowth kineticsMutagenesisProteinsCleavage sitesGenetic engineeringArginineAspergillopepsinFungal proteinFurinGlucan 1,4 alpha glucosidaseHybrid proteinLysineProtein precursorProteinaseRecombinant proteinUnclassified drugAlleleAmino acid compositionAspergillus nigerControlled studyFungal geneFungal strainFungus mutantGene deletionGene disruptionGene functionGenetic codeGenetic recombinationMolecular cloningNonhumanNucleotide sequencePriority journalProtein degradationProtein processingWild typeAspergillus nigerEndopeptidasesFungal ProteinsFurinGene Expression Regulation, EnzymologicGene Expression Regulation, FungalProtein Processing, Post-TranslationalRecombinant Fusion ProteinsTrans-Activation (Genetics)AspergillusAspergillus nigerFungi
TNO Identifier
237524
Repository link
https://resolver.tno.nl/uuid:e8947f3d-9fcf-4d87-8e0f-731229196132
ISSN
01681656
Source
Journal of Biotechnology, 106(1), pp. 23-32.
Pages
23-32
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