cient genetic engineering tools will allow for the
development of cell factories with minimal genomes, where all undesirable cell functions that
do not significantly impact fitness during fermentation have been eliminated. For this purpose, multiplex gene editing has already been
implemented in fungi (Foster et al. 2018; Liu
et al. 2015; Nødvig et al. 2018; Pohl et al. 2016;
Shi et al. 2019). Deletion of a number of BGCs
in A. nidulans with the aim of increasing
the pool of available SM precursors and to
simplify metabolite analysis (Chiang et al.
2013, 2016) already serve as a small scale example of this strategy. More ambitiously, one may
envision that entirely synthetic filamentous
fungi, inspired by the synthetic yeast 2.0 project
(Pretorius and Boeke 2018), may be developed.
Altogether, we conclude that the rapidly
expanding synthetic biology toolbox in combination with the large number of fully sequenced
genomes will set the stage for a very exciting
future of fungal heterologous product production.
Acknowledgments We are grateful to Michael Lynge
Nielsen for carefully reading and editing this book
chapter. This work has been supported by Innovation
Fund Denmark, grant no.: 6150-00031B.
References
Adav SS, Ravindran A, Chao LT, Tan L, Singh S, Sze SK
(2011) Proteomic analysis of pH and strains
dependent protein secretion of Trichoderma reesei. J Proteome Res 10:4579–4596
Adeniran A, Sherer M, Tyo KEJ (2015) Yeast-based
biosensors: design and applications. FEMS Yeast
Res 15:1–15
Adney WS, Chou YC, Decker SR, Ding SY, Baker JO,
Kunkel G, Vinzant TB, Himmel ME (2003) Heterologous expression of Trichoderma reesei 1,4-b-Dglucan cellobiohydrolase (Cel 7A). In: Mansfield
SD, Saddler JN (eds) Applications of enzymes to
lignocellulosics, vol 855. American Chemical Society, Washington, DC, pp 403–437
Adney WS, Jeoh T, Beckham GT, Chou YC, Baker JO,
Michener W, Brunecky R, Himmel ME (2009)
Probing the role of N-linked glycans in the stability and activity of fungal cellobiohydrolases by
mutational analysis. Cellulose 16:699–709
Ahmed SM, Nishida-Fukuda H, Li Y, McDonald WH,
Gradinaru CC, Macara IG (2018) Exocyst dynamics during vesicle tethering and fusion. Nat Commun 9:1–17
Alberti A, Ferrero I, Lodi T (2003) LYS2 gene and
its mutation in Kluyveromyces lactis. Yeast
20:1171–1175
Aleksenko A, Clutterbuck AJ (1997) Autonomous plasmid replication in Aspergillus nidulans: AMA1 and
MATE elements. Fungal Genet Biol 21:373–387
An X, Lu J, Huang J, Zhang B, Liu D, Zhang X, Chen J,
Zhou Y, Tong Y (2007) Rapid assembly of
multiple-exon cDNA directly from genomic DNA.
PLoS One 2:1–7
Andersen MR, Vongsangnak W, Panagiotou G, Salazar
MP, Lehmann L, Nielsen J (2008) A trispecies
Aspergillus microarray: comparative transcriptomics of three Aspergillus species. Proc Natl
Acad Sci USA 105:4387–4392
Andersen MR, Salazar MP, Schaap PJ, Van De Vondervoort PJI, Culley D, Thykaer J, Frisvad JC, Nielsen
KF, Albang R, Albermann K, Berka RM, Braus GH,
Braus-Stromeyer SA, Corrochano LM, Dai Z, Van
Dijck PWM, Hofmann G, Lasure LL, Magnuson JK,
Menke H, Meijer M, Meijer SL, Nielsen JB, Nielsen
ML, Van Ooyen AJJ, Pel HJ, Poulsen L, Samson
RA, Stam H, Tsang A, Van Den Brink JM, Atkins
A, Aerts A, Shapiro H, Pangilinan J, Salamov A,
Lou Y, Lindquist E, Lucas S, Grimwood J, Grigoriev IV, Kubicek CP, Martinez D, van Peij NNME,
Roubos JA, Nielsen J, Baker SE (2011) Comparative genomics of citric-acid-producing Aspergillus
niger ATCC 1015 versus enzyme-producing CBS
513.88. Genome Res 21:885–897
Anyaogu DC, Mortensen UH (2015) Manipulating the
glycosylation pathway in bacterial and lower
eukaryotes for production of therapeutic proteins.
Curr Opin Biotechnol 36:122–128
Archer D, Jeenes D, MacKenzie D, Brightwell G, Lambert N, Lowe G, Radford S, Dobson C (1990) Hen
egg white lysozyme expressed in, and secreted
from, Aspergillus niger is correctly processed and
folded. Biotechnology 8:741–745
Austin B, Hail RM, Tyler BM (1990) Optimized vectors
and selection for transformation of Neurospora
crassa and Aspergillus nidulans to bleomycin and
phleomycin. Gene 93:157–162
Avalos J, Geever RF, Case ME (1989) Bialaphos resistance as a dominant selectable marker in Neurospora crassa. Curr Genet 16:369–372
Barlowe C, Miller EA (2013) Secretory protein biogenesis and traffic in the early secretory pathway.
Genetics 193:383–410
Barredo JL, Cantoral JM, Alvarez E, Dı ´ez B, Martı ´n JF
(1989) Cloning, sequence analysis and transcriptional study of the isopenicillin N synthase of
Penicillium chrysogenum AS-P-78. Mol Gen
Genet 216:91–98
Beck J, Ripka S, Siegner A, Schiltz E, Schweizer E (1990)
The multifunctional 6-methylsalicylic acid
synthase gene of Penicillium patulum. Its gene
structure relative to that of other polyketide
synthases. Eur J Biochem 192:487–498
10 Filamentous Fungi as Hosts for Heterologous Production of Proteins and Secondary. . .
255
development of cell factories with minimal genomes, where all undesirable cell functions that
do not significantly impact fitness during fermentation have been eliminated. For this purpose, multiplex gene editing has already been
implemented in fungi (Foster et al. 2018; Liu
et al. 2015; Nødvig et al. 2018; Pohl et al. 2016;
Shi et al. 2019). Deletion of a number of BGCs
in A. nidulans with the aim of increasing
the pool of available SM precursors and to
simplify metabolite analysis (Chiang et al.
2013, 2016) already serve as a small scale example of this strategy. More ambitiously, one may
envision that entirely synthetic filamentous
fungi, inspired by the synthetic yeast 2.0 project
(Pretorius and Boeke 2018), may be developed.
Altogether, we conclude that the rapidly
expanding synthetic biology toolbox in combination with the large number of fully sequenced
genomes will set the stage for a very exciting
future of fungal heterologous product production.
Acknowledgments We are grateful to Michael Lynge
Nielsen for carefully reading and editing this book
chapter. This work has been supported by Innovation
Fund Denmark, grant no.: 6150-00031B.
References
Adav SS, Ravindran A, Chao LT, Tan L, Singh S, Sze SK
(2011) Proteomic analysis of pH and strains
dependent protein secretion of Trichoderma reesei. J Proteome Res 10:4579–4596
Adeniran A, Sherer M, Tyo KEJ (2015) Yeast-based
biosensors: design and applications. FEMS Yeast
Res 15:1–15
Adney WS, Chou YC, Decker SR, Ding SY, Baker JO,
Kunkel G, Vinzant TB, Himmel ME (2003) Heterologous expression of Trichoderma reesei 1,4-b-Dglucan cellobiohydrolase (Cel 7A). In: Mansfield
SD, Saddler JN (eds) Applications of enzymes to
lignocellulosics, vol 855. American Chemical Society, Washington, DC, pp 403–437
Adney WS, Jeoh T, Beckham GT, Chou YC, Baker JO,
Michener W, Brunecky R, Himmel ME (2009)
Probing the role of N-linked glycans in the stability and activity of fungal cellobiohydrolases by
mutational analysis. Cellulose 16:699–709
Ahmed SM, Nishida-Fukuda H, Li Y, McDonald WH,
Gradinaru CC, Macara IG (2018) Exocyst dynamics during vesicle tethering and fusion. Nat Commun 9:1–17
Alberti A, Ferrero I, Lodi T (2003) LYS2 gene and
its mutation in Kluyveromyces lactis. Yeast
20:1171–1175
Aleksenko A, Clutterbuck AJ (1997) Autonomous plasmid replication in Aspergillus nidulans: AMA1 and
MATE elements. Fungal Genet Biol 21:373–387
An X, Lu J, Huang J, Zhang B, Liu D, Zhang X, Chen J,
Zhou Y, Tong Y (2007) Rapid assembly of
multiple-exon cDNA directly from genomic DNA.
PLoS One 2:1–7
Andersen MR, Vongsangnak W, Panagiotou G, Salazar
MP, Lehmann L, Nielsen J (2008) A trispecies
Aspergillus microarray: comparative transcriptomics of three Aspergillus species. Proc Natl
Acad Sci USA 105:4387–4392
Andersen MR, Salazar MP, Schaap PJ, Van De Vondervoort PJI, Culley D, Thykaer J, Frisvad JC, Nielsen
KF, Albang R, Albermann K, Berka RM, Braus GH,
Braus-Stromeyer SA, Corrochano LM, Dai Z, Van
Dijck PWM, Hofmann G, Lasure LL, Magnuson JK,
Menke H, Meijer M, Meijer SL, Nielsen JB, Nielsen
ML, Van Ooyen AJJ, Pel HJ, Poulsen L, Samson
RA, Stam H, Tsang A, Van Den Brink JM, Atkins
A, Aerts A, Shapiro H, Pangilinan J, Salamov A,
Lou Y, Lindquist E, Lucas S, Grimwood J, Grigoriev IV, Kubicek CP, Martinez D, van Peij NNME,
Roubos JA, Nielsen J, Baker SE (2011) Comparative genomics of citric-acid-producing Aspergillus
niger ATCC 1015 versus enzyme-producing CBS
513.88. Genome Res 21:885–897
Anyaogu DC, Mortensen UH (2015) Manipulating the
glycosylation pathway in bacterial and lower
eukaryotes for production of therapeutic proteins.
Curr Opin Biotechnol 36:122–128
Archer D, Jeenes D, MacKenzie D, Brightwell G, Lambert N, Lowe G, Radford S, Dobson C (1990) Hen
egg white lysozyme expressed in, and secreted
from, Aspergillus niger is correctly processed and
folded. Biotechnology 8:741–745
Austin B, Hail RM, Tyler BM (1990) Optimized vectors
and selection for transformation of Neurospora
crassa and Aspergillus nidulans to bleomycin and
phleomycin. Gene 93:157–162
Avalos J, Geever RF, Case ME (1989) Bialaphos resistance as a dominant selectable marker in Neurospora crassa. Curr Genet 16:369–372
Barlowe C, Miller EA (2013) Secretory protein biogenesis and traffic in the early secretory pathway.
Genetics 193:383–410
Barredo JL, Cantoral JM, Alvarez E, Dı ´ez B, Martı ´n JF
(1989) Cloning, sequence analysis and transcriptional study of the isopenicillin N synthase of
Penicillium chrysogenum AS-P-78. Mol Gen
Genet 216:91–98
Beck J, Ripka S, Siegner A, Schiltz E, Schweizer E (1990)
The multifunctional 6-methylsalicylic acid
synthase gene of Penicillium patulum. Its gene
structure relative to that of other polyketide
synthases. Eur J Biochem 192:487–498
10 Filamentous Fungi as Hosts for Heterologous Production of Proteins and Secondary. . .
255