Abstract
Despite the naturally high capacity for protein secretion by many species of filamentous fungi, secteted yields of many heterologous proteins have been comparatively low. The strategies for yield improvement have included the use of strong homologous promoters, increased gene copy number, gene fusions with a gene encoding a naturally well-secreted protein, protease-deficient host strains and screening for high yields following random mutagenesis. Such approaches have been effective with some target heterologous proteins but not others.
Approaches used in heterologous protein production from filamentous fungi are discussed and a perspective on emerging strategies is presented.
Similar content being viewed by others
References
Archer DB, Jeenes DJ, MacKenzie DA, Brightwell G, Lambert N, Lowe G, Radford SE & Dobson CM (1990) Hen egg white lysozyme expressed in, and secreted from,Aspergillus niger is correctly processed and folded. Biotechnol. 8: 741–745
Archer DB, MacKenzie DA, Jeenes DJ & Roberts IN (1992) Proteolytic degradation of heterologous proteins expressed inAspergillus niger. Biotechnol. Lett. 14: 357–362
Berka RM, Ward M, Wilson LJ, Hayenga KJ, Kodama KH, Carlomagno LP & Thompson SA (1990) Molecular cloning and deletion of the aspergillopepsin A gene fromAspergillus awamori. Gene 86: 153–162
Calmels TPG, Martin F, Durand H & Tiraby G (1991) Proteolytic events in the processing of secreted proteins in fungi. J. Biotechnol. 17: 51–66
Contreras R, Carrez D, Kinghorn JR, Van den Hondel CAMJJ & Fiers W (1991) Efficient KEX2-like processing of a glucoamylase-interleukin-6 fusion protein byAspergillus nidulans and secretion of mature interleukin-6. Biotechnol. 9: 378–381
Cullen D, Gray GL, Wilson LJ, Hayenga KJ, Lamsa MH, Rey MW, Norton S & Berka RM (1987) Controlled expression and secretion of bovine chymosin inAspergillus nidulans. Biotechnol. 5: 369–376
Davies RW (1991) Molecular biology of a high-level recombinant protein production system inAspergillus. In: Leong SA & Berka RM (Eds) Molecular Industrial Mycology (pp 45–81). Marcel Dekker, New York
Dunn-Coleman NS, Bloebaum P, Berka RM, Bodie E, Robinson N, Armstrong G, Ward M, Przetak M, Carter G, LaCost R, Wilson LJ, Kodama KH, Baliu EF, Bower B, Lamsa M & Heinsohn H (1991) Commercially viable levels of chymosin production byAspergillus. Biotechnol. 9: 976–981
Fowler T, Berka RM & Ward M (1990) Regulation of theglaA gene ofAspergillus niger. Curr. Genet. 18: 537–545
Gwynne DI, Buxton FP, Williams SA, Garven S & Davies RW (1987) Genetically engineered secretion of active human interferon and a bacterial endoglucanase fromAspergillus nidulans. Biotechnol. 5: 713–719
Harkki A, Uusitalo J, Bailey M, Penttila M & Knowles JKC (1989) A novel fungal expression system: Secretion of active calf chymosin from the filamentous fungusTrichoderma reesei. Biotechnol. 7: 596–603
Hodgson J (1993) Expression systems: A user's guide. Bio/technology 11: 887–893
Jeenes DJ, MacKenzie DA, Roberts IN & Archer DB (1991) Heterologous protein production by filamentous fungi. Biotechnol. Genet. Eng. Rev. 9: 327–367
Jeenes DJ, Marczinke B, MacKenzie DA & Archer DB (1993) A truncated glucoamylase gene fusion for heterologous protein secretion fromAspergillus niger. FEMS Microbiol. Lett. 107: 267–272
Lumb KJ, Aplin RT, Radford SE, Archer DB, Jeenes DJ, Lambert N, MacKenzie DA, Dobson CM & Lowe G (1992) A study of D52S hen lysozyme-GlcNAc oligosaccharide complexes by NMR spectroscopy and electrospray mass spectrometry. FEBS Lett. 296: 153–157
MacKenzie DA, Jeenes DJ, Belshaw NJ & Archer DB (1993a) Regulation of secreted protein production by filamentous fungi: Recent developments and perspectives. J. Gen. Microbiol. 139: 2295–2307
MacKenzie DA, Gendron LCG, Jeenes DJ & Archer DB (1993b) Physiological optimisation of secreted protein production byAspergillus niger. Enz. Microb. Technol., in press
Mattern IE, Van Noort JM, Van den Berg P, Archer DB, Roberts IN & Van den Hondel CAMJJ (1992) Isolation and characterization of mutants ofAspergillus niger deficient in extracellular proteases. Mol. Gen. Genet. 234: 332–336
Nakano ET, Fox RD, Clements DE, Woo K, Stuart WD & Ivy JM (1993) Expression vectors forNeurospora crassa and expression of a bovine preprochymosin cDNA. Fung. Genet. Newslett. 40: 54–56
Nyyssönen E, Penttilä M, Harkki A, Saloheimo A, Knowles JKC & Keränen S (1993) Efficient production of antibody fragments by the filamentous fungusTrichoderma reesei. Biotechnol. 11: 591–595
Roberts IN, Jeenes DJ, MacKenzie DA, Wilkinson AP, Sumner IG & Archer DB (1992a) Heterologous gene expression inAspergillus niger: A glucoamylase-porcine pancreatic prophospholipase A2 fusion protein is secreted and processed to yield mature enzyme. Gene 122: 155–161
Roberts IN, MacKenzie DA, Jeenes DJ, Archer DB, Radford SE, Robinson CV, Aplin RT & Dobson CM (1992b) Production of15N-labelled hen egg white lysozyme usingAspergillus niger. Biotechnol. Lett. 14: 897–902
Van den Hondel CAMJJ, Punt PJ & Van Gorcom RFM (1991) Heterologous gene expression in filamentous fungi. In: Bennett JW & Lasure LL (Eds) More Genetic Manipulation of Filamentous Fungi (pp 396–428). Academic Press, Orlando
Van Hartingsveldt W, Van Zeijl CMJ, Veenstra AE, Van den Berg JA, Pouwels PH, Van Gorcom RFM & Van den Hondel CAMJJ (1990) Heterologous gene expression inAspergillus: analysis of chymosin production in single-copy transformants ofAspergillus niger. In: Heslot H, Davies J, Florent J, Bibichon L, Durant G & Penasse L (Eds) Proceedings of 6th International Symposium on Genetics of Industrial Microorganisms (pp. 107–116) (Société Française de Microbiologie), Strassbourg
Verdoes JC, Punt PJ, Schrickx JM, Van Verseveld HW, Stouthamer AH & Van den Hondel CAMJJ (1993) Glucoamylase overexpression inAspergillus niger: Molecular genetic analysis of strains containing multiple copies of theglaA gene. Transgenic Res. 2: 84–92
Ward M (1991) Chymosin production inAspergillus. In: Leong SA & Berka RM (Eds) Molecular Industrial Mycology (pp 83–105). Marcel Dekker, New York
Ward M, Wilson LJ & Kodama KH (1993) Use of Aspergillus over-producing mutants, cured for integrated plasmid, to overproduce heterologous proteins. Appl. Microbiol. Biotechnol. 39: 738–743
Ward M, Wilson LJ, Kodama KH, Rey MW & Berka RM (1990) Improved production of chymosin inAspergillus by expression as a glucoamylase-chymosin fusion. Biotechnol. 8: 435–440
Williamson G, Belshaw NJ & Williamson MP (1992) O-glycosylation in Aspergillus glucoamylase: conformation and role in binding. Biochem. J. 282: 423–428
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Archer, D.B., Jeenes, D.J. & Mackenzie, D.A. Strategies for improving heterologous protein production from filamentous fungi. Antonie van Leeuwenhoek 65, 245–250 (1994). https://doi.org/10.1007/BF00871952
Issue Date:
DOI: https://doi.org/10.1007/BF00871952