Abstract
Aspergillus niger is a genetically tractable model organism for scientific discovery and a platform organism used for the production of heterologous enzymes. Academic strategies to increase production include the use of strong promoters, multiple gene copies, gene knock out and knock in strains, secretion pathway engineering, and multi omics-based approaches. However, yields of heterologous proteins are still lower than desired. Industrial approaches are more effective and straightforward: optimizing bioprocess conditions, strains engineering through multiple rounds of mutagenesis and reverse genetics, and new expression constructs design and testing, which are based on high-throughput screening in automated bioreactor/fermenter. High-level production of heterologous enzymes remains elusive in Aspergillus niger but more promising than ever.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Adav SS, Ravindran A et al (2013) Proteomic analysis of temperature dependent extracellular proteins from Aspergillus fumigatus grown under solid-state culture condition. J Proteome Res 12(6):2715–2731
Al-Sheikh H, Watson AJ et al (2004) Endoplasmic reticulum stress leads to the selective transcriptional downregulation of the glucoamylase gene in Aspergillus niger. Mol Microbiol 53(6):1731
Beckham GT, Dai Z et al (2012) Harnessing glycosylation to improve cellulase activity. Curr Opin Biotechnol 23(3):338–345
Bhat MK (2000) Cellulases and related enzymes in biotechnology. Biotechnol Adv 18(5):355
Blumhoff M, Steiger MG et al (2013) Six novel constitutive promoters for metabolic engineering of Aspergillus niger. Appl Microbiol Biotechnol 97(1):259–267
Broekhuijsen MP, Mattern IE et al (1993) Secretion of heterologous proteins by Aspergillus niger : production of active human interleukin-6 in a protease-deficient mutant by KEX2-like processing of a glucoamylase-hIL6 fusion protein. J Biotechnol 31(2):135–145
Conesa A, Punt PJ et al (2001) The secretion pathway in filamentous fungi: a biotechnological view. Fungal Genet Biol 33(3):155–171
Davies J, Jiang L et al (2001) Expression of GnTIII in a recombinant anti-CD20 CHO production cell line: expression of antibodies with altered glycoforms leads to an increase in ADCC through higher affinity for FC gamma RIII. Biotechnol Bioeng 74(4):288
Deshpande N, Wilkins MR et al (2008) Protein glycosylation pathways in filamentous fungi. Glycobiology 18(8):626
Egrie JC, Browne JK (2001) Development and characterization of novel erythropoiesis stimulating protein (NESP). Br J Cancer Suppl 1:3–10
Enshasy E (2007) Filamentous fungal cultures-process characteristics, products, and applications. In: Bioprocessing for value-added products from renewable resources. Elsevier, London
Fleissner A, Dersch P (2010) Expression and export: recombinant protein production systems for Aspergillus. Appl Microbiol Biotechnol 87(4):1255–1270
Gao L, Gao F et al (2012) N-Glycoform diversity of cellobiohydrolase I from Penicillium decumbens and synergism of nonhydrolytic glycoform in cellulose degradation. J Biol Chem 287(19):15906–15915
Gerngross TU (2004) Advances in the production of human therapeutic proteins in yeasts and filamentous fungi. Nat Biotechnol 22(11):1409
Gilbert JA, Dupont CL (2011) Microbial metagenomics: beyond the genome. Annu Rev Mar Sci 3(1):347–371
Gladden JM, Park JI et al (2014) Discovery and characterization of ionic liquid-tolerant thermophilic cellulases from a switchgrass-adapted microbial community. Biotechnol Biofuels 7(1):15
Gouka RJ, Punt PJ et al (1997a) Efficient production of secreted proteins by Aspergillus: progress, limitations and prospects. Appl Microbiol Biotechnol 47(1):1–11
Gouka RJ, Punt PJ et al (1997b) Glucoamylase gene fusions alleviate limitations for protein production in Aspergillus awamori at the transcriptional and (post) translational levels. Appl Environ Microbiol 63(2):488
Guillemette T, Peij NNV et al (2007) Genomic analysis of the secretion stress response in the enzyme-producing cell factory Aspergillus niger. BMC Genomics 8(1):158
Gupta VK (2016) Fungal enzymes for bio-products from sustainable and waste biomass. Trends Biochem Sci 41(7):633–645
Hijarrubia MJ, Casqueiro J et al (1997) Characterization of the bip gene of Aspergillus awamori encoding a protein with an HDEL retention signal homologous to the mammalian BiP involved in polypeptide secretion. Curr Genet 32(2):139
Hombergh JPTW, Gelpke MDS et al (1997) Disruption of three acid proteases in Aspergillus niger – effects on protease spectrum, intracellular proteolysis, and degradation of target proteins. Eur J Biochem 247(2):605
Inoue H, Kimura T et al (1991) The gene and deduced protein sequences of the zymogen of Aspergillus niger acid proteinase A. J Biol Chem 266(29):19484
Jeenes DJ, Pfaller R et al (1997) Isolation and characterisation of a novel stress-inducible PDI-family gene from Aspergillus niger. Gene 193(2):151–156
Jin C (2012) Protein glycosylation in Aspergillus fumigatus is essential for cell wall synthesis and serves as a promising model of multicellular eukaryotic development. Int J Microbiol 2012(1687-918X):654251
Joosten V, Lokman C et al (2003) The production of antibody fragments and antibody fusion proteins by yeasts and filamentous fungi. Microb Cell Factories 2(1):1–15
Kleizen B, Braakman I (2004) Protein folding and quality control in the endoplasmic reticulum. Curr Opin Cell Biol 16(4):343
Krishnan S, Vijayalakshmi MA (1985) Purification of an acid protease and a serine carboxypeptidase from Aspergillus niger using metal-chelate affinity chromatography. J Chromatogr 329(1):165
Lam PVN, Goldman R et al (2013) Structure-based comparative analysis and prediction of N-linked glycosylation sites in evolutionarily distant eukaryotes. Genomics Proteomics Bioinformatics 11(2):96–104
Linden H, Kaushansky K (2000) The glycan domain of thrombopoietin enhances its secretion. Biochemistry 39(11):3044
Lotfy WA, Ghanem KM et al (2007) Citric acid production by a novel Aspergillus niger isolate: I. Mutagenesis and cost reduction studies. Bioresour Technol 98(18):3464
Lubertozzi D, Keasling JD (2009) Developing Aspergillus as a host for heterologous expression. Biotechnol Adv 27(1):53
Mattern IE, Noort JMV et al (1992) Isolation and characterization of mutants of Aspergillus niger deficient in extracellular proteases. Mol Gen Genet 234(2):332–336
Meyer V (2008) Genetic engineering of filamentous fungi – progress, obstacles and future trends. Biotechnol Adv 26(2):177–185
Meyer V, Bo W et al (2011) Aspergillus as a multi-purpose cell factory: current status and perspectives. Biotechnol Lett 33(3):469
Meyer V, Fiedler M et al (2015) The cell factory Aspergillus enters the big data era: opportunities and challenges for optimising product formation. Adv Biochem Eng Biotechnol 149:91
Mikko A, Tiina P et al (2006) Common features and interesting differences in transcriptional responses to secretion stress in the fungi Trichoderma reesei and Saccharomyces cerevisiae. BMC Genomics 7(1):32
Mistry PK, Wraight EP et al (1996) Therapeutic delivery of proteins to macrophages: implications for treatment of Gaucher’s disease. Lancet 348(9041):1555–1559
Mizutani O, Masaki K et al (2012) Modified Cre-loxP recombination in Aspergillus oryzae by direct introduction of Cre recombinase for marker gene rescue. Appl Environ Microbiol 78(12):4126–4133
Mulder HJ, Saloheimo M et al (2004) The transcription factor HACA mediates the unfolded protein response in Aspergillus niger, and up-regulates its own transcription. Mol Gen Genomics 271(2):130–140
Nevalainen H, Peterson R (2014) Making recombinant proteins in filamentous fungi- are we expecting too much? Front Microbiol 5(1):75
Nevalainen KMH, Te’O VSJ et al (2005) Heterologous protein expression in filamentous fungi. Trends Biotechnol 23(9):468–474
Nødvig CS, Nielsen JB et al (2015) A CRISPR-Cas9 system for genetic engineering of filamentous fungi. PLoS One 10(7):e0133085
Ogundero VW (1987) Temperature and aflatoxin production by Aspergillus flavus and A. parasiticus strains from Nigerian groundnuts. J Basic Microbiol 27(9):511–514
Park JI, Steen EJ et al (2012) A thermophilic ionic liquid-tolerant cellulase cocktail for the production of cellulosic biofuels. PLoS One 7(5):e37010
Pel HJ, Winde JHD et al (2007) Genome sequencing and analysis of the versatile cell factory Aspergillus niger CBS 513.88. Nat Biotechnol 25(2):221
Pérez J, Muñozdorado J et al (2002) Biodegradation and biological treatments of cellulose, hemicellulose and lignin: an overview. Int Microbiol 5(2):53–63
Peterson R, Nevalainen H (2012) Trichoderma reesei RUT-C30 – thirty years of strain improvement. Microbiology 158(1):58–68
Quinlan RJ, Sweeney MD et al (2011) Insights into the oxidative degradation of cellulose by a copper metalloenzyme that exploits biomass components. Proc Natl Acad Sci U S A 108(37):15079–15084
Rab MA, Freeman TW et al (2013) Integrated approaches for assessment of cellular performance in industrially relevant filamentous fungi. Ind Biotechnol 9(6):337–344
Ries LN, Beattie SR et al (2016) Diverse regulation of the CreA carbon catabolite repressor in Aspergillus nidulans. Genetics 203(1):335–352
Ruijter GJ, Vanhanen SA et al (1997) Isolation of Aspergillus niger creA mutants and effects of the mutations on expression of arabinases and L-arabinose catabolic enzymes. Microbiology 143(Pt 9):2991
Ruiz-DÃez B (2002) A review: strategies for the transformation of filamentous fungi. J Appl Microbiol 92(2):189
Saloheimo M, Valkonen M et al (2003) Activation mechanisms of the HAC1-mediated unfolded protein response in filamentous fungi. Mol Microbiol 47(4):1149–1161
Schubert C (2006) Can biofuels finally take center stage? Nat Biotechnol 24(7):777
Sims AH, Gent ME et al (2005) Transcriptome analysis of recombinant protein secretion by Aspergillus nidulans and the unfolded-protein response in vivo. Appl Environ Microbiol 71(5):2737–2747
Stals I, Sandra K et al (2004) Factors influencing glycosylation of Trichoderma reesei cellulases. I: postsecretorial changes of the O- and N-glycosylation pattern of Cel7A. Glycobiology 14(8):713–724
Tadesse H, Luque R (2011) Advances on biomass pretreatment using ionic liquids: an overview. Energy Environ Sci 4(10):3913–3929
Thompson SA (1990) Molecular cloning and deletion of the gene encoding aspergillopepsin A from Aspergillus awamori. Gene 86(2):153–162
Travers KJ, Patil CK et al (2000) Functional and genomic analyses reveal an essential coordination between the unfolded protein response and ER-associated degradation. Cell 101(3):249–258
Valkonen M, Ward M et al (2003) Improvement of foreign-protein production in Aspergillus niger var. awamori by constitutive induction of the unfolded-protein response. Appl Environ Microbiol 69(12):6979–6986
van den Berg BA, Reinders MJ et al (2012) Exploring sequence characteristics related to high-level production of secreted proteins in Aspergillus niger. PLoS One 7(10):e45869
van den Berg BA, Reinders MJ et al (2014) Protein redesign by learning from data. Protein Eng Des Sel 27(9):281–288
van den Hombergh JP, Jarai G et al (1994) Cloning, characterization and expression of pepF, a gene encoding a serine carboxypeptidase from Aspergillus niger. Gene 151(1–2):73–79
van den Hombergh JP, van de Vondervoort PJ et al (1997) Aspergillus as a host for heterologous protein production: the problem of proteases. Trends Biotechnol 15(7):256
Van Dyk JS, Pletschke BI (2012) A review of lignocellulose bioconversion using enzymatic hydrolysis and synergistic cooperation between enzymes-Factors affecting enzymes, conversion and synergy. Biotechnol Adv 30(6):1458–1480
Varki A, Cummings RD et al (2009) Part I, general principles, essentials of glycobiology, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
Verdoes JC, Punt PJ et al (1994) The effect of multiple copies of the upstream region on expression of the Aspergillus niger glucoamylase-encoding gene. Gene 145(2):179–187
Ward M, Wilson LJ et al (1990) Improved production of chymosin in Aspergillus by expression as a glucoamylase-chymosin fusion. Nat Biotechnol 8(5):435–440
Wilson DB (2009) Cellulases and biofuels. Curr Opin Biotechnol 20(3):295–299
Zhang J, Mao Z et al (2011) Ku80 gene is related to non-homologous end-joining and genome stability in Aspergillus niger. Curr Microbiol 62(4):1342–1346
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Nature Singapore Pte Ltd.
About this chapter
Cite this chapter
Zhang, J., Huang, Y., Wang, H. (2018). Heterologous Expression of Lignocellulolytic Enzymes in Aspergillus niger . In: Fang, X., Qu, Y. (eds) Fungal Cellulolytic Enzymes. Springer, Singapore. https://doi.org/10.1007/978-981-13-0749-2_8
Download citation
DOI: https://doi.org/10.1007/978-981-13-0749-2_8
Published:
Publisher Name: Springer, Singapore
Print ISBN: 978-981-13-0748-5
Online ISBN: 978-981-13-0749-2
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)