Evaluation of Aspergillus niger as host for virus-like particle production, using the hepatitis B surface antigen as a model
van Zyl W.H.
The filamentous fungus Aspergillus niger was transformed with the hepatitis B virus S gene encoding the major viral envelope protein under control of the constitutive A. nidulans glyceraldehyde-3-phosphate dehydrogenase (gpdA) promoter. Approximately seven copies of the expression cassette were integrated on the genome, resulting in high-level transcription of the S gene. Production of the 24-kDa S protein and a 48-kDa S protein dimer in the membrane-associated protein fraction of the recombinant A. niger strain was shown through Western analysis. Electron microscopy of partially purified recombinant S protein revealed the formation of spherical pseudoviral particles with a diameter of 22 nm. The production level of hepatitis B pseudoviral particles was estimated to be 0.4 mg/1 culture, which compares favourably with the reported levels initially obtained in yeast, indicating the potential of the Aspergillus expression system as an alternative, cost-effective vaccine production system.
dimer, glyceraldehyde 3 phosphate dehydrogenase, hepatitis B surface antigen, recombinant protein, virus envelope protein, vitronectin, article, Aspergillus nidulans, Aspergillus niger, cost effectiveness analysis, electron microscopy, fungal strain, fungus culture, gene cassette, gene expression, genetic transcription, genetic transformation, genome, Hepatitis B virus, host, nonhuman, particle size, priority journal, promoter region, protein purification, protein synthesis, vaccine production, virus gene, virus particle, Western blotting, Aspergillus niger, Bioreactors, Biotechnology, Gene Dosage, Genetic Engineering, Hepatitis B Surface Antigens, Hepatitis B virus, Humans, Models, Genetic, Promoter Regions (Genetics), Transformation, Genetic, Transgenes, Viral Proteins, Virion, Aspergillus, Aspergillus niger, Emericella nidulans, Fungi, Hepatitis B virus