Identification of a classical mutant in the industrial host Aspergillus niger by systems genetics: LaeA is required for citric acid production and regulates the formation of some secondary metabolites

G3: Genes, Genomes, Genetics

Volumn 6, Issue 1, 2016, Pages 193-204

  Niu, Jing ^a   Arentshorst, Mark ^a   Nair, P Deepa S ^a   Dai, Ziyu ^b   Baker, Scott E ^c   Frisvad, Jens C ^d   Nielsen, Kristian F ^d   Punt, Peter J ^a,e   Ram, Arthur F J ^a  

^a LEIDEN UNIVERSITY   (Netherlands)

^b PACIFIC NORTHWEST NATIONAL LABORATORY   (United States)

^c PACIFIC NORTHWEST NATIONAL LABORATORY   (United States)

^d TECHNICAL UNIVERSITY OF DENMARK   (Denmark)

^e Dutch DNA Biotech   (Netherlands)

Author keywords

Bulk segregant analysis; Filamentous fungi; Genome sequencing; Organic acids; Parasexual cycle

Indexed keywords

ASPERNIGRIN A; ASPERNIGRIN B; ASPERRUBROL; ATROMENTIN; AURASPERONE B; CITRIC ACID; DEMETHYLKOTANIN; FUMONISIN B2; FUMONISIN B4; FUNALENONE; FUNGAL PROTEIN; FUNGISPORIN A; GENOMIC DNA; KOTANIN; NIGERAZINE; NIGRAGILIN; ORLANDIN; OXALIC ACID; PESTALAMIDE C; PYRANONIGRIN S; PYRANOPYRROL A; PYROPHEN; TENSIDOL B; UNCLASSIFIED DRUG;

ARTICLE; ASPERGILLUS NIGER; AUXOTROPHY; BULK SEGREGANT ANALYSIS; CONTROLLED STUDY; DIPLOIDY; DNA SEQUENCE; ENZYME ACTIVITY; ENZYME SYNTHESIS; FUNGAL GENE; FUNGAL STRAIN; FUNGUS GROWTH; FUNGUS MUTANT; GENE EXPRESSION; GLUCOSE INTAKE; HIGH PERFORMANCE LIQUID CHROMATOGRAPHY; HIGH THROUGHPUT SEQUENCING; LAEA GENE; NEXT GENERATION SEQUENCING; NONHUMAN; PHENOTYPE; POINT MUTATION; SEGREGATION ANALYSIS; ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY; ALLELE; GENETIC COMPLEMENTATION; GENETICS; GENOTYPE; ISOLATION AND PURIFICATION; METABOLISM; MUTATION; SECONDARY METABOLISM; SINGLE NUCLEOTIDE POLYMORPHISM;

ALLELES; ASPERGILLUS NIGER; CITRIC ACID; GENES, FUNGAL; GENETIC COMPLEMENTATION TEST; GENOTYPE; MUTATION; PHENOTYPE; POLYMORPHISM, SINGLE NUCLEOTIDE; SECONDARY METABOLISM;

EID: 84953791222     PISSN: None     EISSN: 21601836     Source Type: Journal    
DOI: 10.1534/g3.115.024067     Document Type: Article

References (86)

1. Andersen, M.R., L. Lehmann, and J. Nielsen, 2009 Systemic analysis of the response of Aspergillus niger to ambient pH. Genome Biol. 10: R47.
2. Andersen, M.R., M.P. Salazar, P.J. Schaap, P.J. I. van de Vondervoort, D. Culley et al. 2011 Comparative genomics of citric-acid-producing Aspergillus niger ATCC 1015 vs. enzyme-producing CBS 513.88. Genome Res. 21: 885-897.
3. Andersen, M.R., J.B. Nielsen, A. Klitgaard, L.M. Petersen, M. Zachariasen et al. 2013 Accurate prediction of secondary metabolite gene clusters in filamentous fungi. Proc. Natl. Acad. Sci. USA 110: E99-E107.
4. Arentshorst, M., A.F. J. Ram, and V. Meyer, 2012 Using non-homologous end-joining-deficient strains for functional gene analyses in filamentous fungi. Methods Mol. Biol. 835: 133-150.
5. Arentshorst, M., J. Niu, and A. F. Ram, 2015 Efficient generation of Aspergillus niger knock out strains by combining NHEJ mutants and a split marker approach, pp. 263-272 in Genetic Transformation Systems in Fungi, Vol. 1, edited by van den Berg, M.A., and K. Maruthachalam. Springer International Publishing, Switzerland.
6. Baker, S.E., 2006 Aspergillus niger genomics: past present and into the future. Med. Mycol. 44(Suppl 1): S17-S21.
7. Bayram, O., and G. H. Braus, 2012 Coordination of secondary metabolism and development in fungi: the velvet family of regulatory proteins. FEMS Microbiol. Rev. 36: 1-24.
8. Benghazi, L., E. Record, A. Suárez, J.A. Gomez-Vidal, J. Martínez et al. 2014 Production of the Phanerochaete flavido-alba laccase in Aspergillus niger for synthetic dyes decolorization and biotransformation. World J. Microbiol. Biotechnol. 30: 201-211.
9. Benoit, I., M. Asther, Y. Bourne, D. Navarro, S. Canaan et al. 2007 Gene overexpression and biochemical characterization of the biotechnologically relevant chlorogenic acid hydrolase from Aspergillus niger. Appl. Environ. Microbiol. 73: 5624-5632.
10. Bennett, J.W., and L. L. Lasure, 1991 Growth media, pp. 441-458 in More Gene Manipulations in Fungi, edited by Bennett, J.W., and L. L. Lasure. Academic Press, San Diego.
11. Bergmeyer, H.U., 1985 Metabolites 2: tri- and dicarboxylic acids, purines, pyrimidines and derivatives, coenzymes, inorganic compounds, pp. 5-10 in Citric Acid, edited by Bergmeyer, H.U., and J. Bergmeyer, and Grassl, M. VCH.
12. Bok, J.W., and N. P. Keller, 2004 2006, LaeA, a regulator of secondary metabolism in Aspergillus spp. Eukaryot. Cell 3: 527-535.
13. Bok, J.W., D. Noordermeer, S.P. Kale, and N. P. Keller, 2006 Secondary metabolic gene cluster silencing in Aspergillus nidulans. Mol. Microbiol. 61: 1636-1645.
14. Bok, J.W., Y.-M. Chiang, E. Szewczyk, Y. Reyes-Dominguez, A.D. Davidson et al. 2009 Chromatin-level regulation of biosynthetic gene clusters. Nat. Chem. Biol. 5: 462-464.
15. Bok, J.W., P. Wiemann, G.S. Garvey, F.Y. Lim, B. Haas et al. 2014 Illumina identification of RsrA, a conserved C2H2 transcription factor coordinating the NapA mediated oxidative stress signaling pathway in Aspergillus. BMC Genomics 15: 1011.
16. Bos, C.J., A.J. Debets, K. Swart, A. Huybers, G. Kobus et al. 1988 Genetic analysis and the construction of master strains for assignment of genes to six linkage groups in Aspergillus niger. Curr. Genet. 14: 437-443.
17. Bos, C.J., S.M. Slakhorst, and A. J. M. Debets, and Swart, K., 1993 Linkage group analysis in Aspergillus niger. Appl. Microbiol. Biotechnol. 38: 742-745.
18. Braaksma, M., and P. J. Punt, 2008 Aspergillus as a cell factory for protein production: controlling protease activity in fungal production, pp. 441-455 in The Aspergilli: Genomics, Medical Aspects, Biotechnology, and Research Methods, edited by Goldman, G.H., and S. A. Osmani. CRC Press, Boca Raton.
19. Braaksma, M., A.K. Smilde, M.J. van der Werf, and P. J. Punt, 2009 The effect of environmental conditions on extracellular protease activity in controlled fermentations of Aspergillus niger. Microbiology 155: 3430-3439.
20. Brown, D.W., R.A. E. Butchko, M. Busman, and R. H. Proctor, 2007 The Fusarium verticillioides FUM gene cluster encodes a Zn(II)2Cys6 protein that affects FUM gene expression and fumonisin production. Eukaryot. Cell 6: 1210-1218.
21. Butchko, R.A. E., D.W. Brown, M. Busman, B. Tudzynski, and P. Wiemann, 2012 Lae1 regulates expression of multiple secondary metabolite gene clusters in Fusarium verticillioides. Fungal Genet. Biol. 49: 602-612.
22. Carvalho, N.D. S. P., M. Arentshorst, M.-J. Kwon, V. Meyer, and A. F. J. Ram, 2010 Expanding the ku70 toolbox for filamentous fungi: establishment of complementation vectors and recipient strains for advanced gene analyses. Appl. Microbiol. Biotechnol. 87: 1463-1473.
23. Chiang, Y.-M., K.M. Meyer, M. Praseuth, S.E. Baker, K.S. Bruno et al. 2011 Characterization of a polyketide synthase in Aspergillus niger whose product is a precursor for both dihydroxynaphthalene (DHN) melanin and naphtho-g-pyrone. Fungal Genet. Biol. 48: 430-437.
24. Chimphango, A.F. A., S.H. Rose, W.H. van Zyl, and J. F. Görgens, 2012 Production and characterisation of recombinant a-Larabinofuranosidase for production of xylan hydrogels. Appl. Microbiol. Biotechnol. 95: 101-112.
25. Dai, Z., and S. Baker, 2015 Enhanced citric acid production in Aspergillus with inactivated asparagine-linked glycosylation protein 3 (Alg3), and/or increased laeA expression. US 9,023,637 B2.
26. Dai, Z., X. Mao, J.K. Magnuson, and L. L. Lasure, 2004 Identification of genes associated with morphology in Aspergillus niger by using suppression subtractive hybridization. Appl. Environ. Microbiol. 70: 2474-2485.
27. Damveld, R.A., A. Franken, M. Arentshorst, P.J. Punt, F.M. Klis et al. 2008 A novel screening method for cell wall mutants in Aspergillus niger identifies UDP-galactopyranose mutase as an important protein in fungal cell wall biosynthesis. Genetics 178: 873-881.
28. de Ruiter-Jacobs, Y.M., M. Broekhuijsen, S.E. Unkles, E.I. Campbell, J.R. Kinghorn et al. 1989 A gene transfer system based on the homologous pyrG gene and efficient expression of bacterial genes in Aspergillus oryzae. Curr. Genet. 16: 159-163.
29. Dunham, M.J., 2012 Two flavors of bulk segregant analysis in yeast. Methods Mol. Biol. 871: 41-54.
30. Fraissinet-Tachet, L., J.P. T. W. van den Hombergh, P.J. van de Vondervoort, G. Jarai, and J. Visser, 1996 Complex regulation of extracelluar proteases in Aspergillus niger; an analysis of wide domain regulatory mutants demonstrates CreA, AreA and pacC control. Thesis, van den. Hombergh, J.P. T. W. Wageningen University. Available at: http:// edepot.wur.nl/200496.
31. Gordon, C.L., V. Khalaj, A.F. Ram, D.B. Archer, J.L. Brookman et al. 2000 Glucoamylase:green fluorescent protein fusions to monitor protein secretion in Aspergillus niger. Microbiology 146: 415-426.
32. Inoue, H., H. Nojima, and H. Okayama, 1990 High efficiency transformation of Escherichia coli with plasmids. Gene 96: 23-28.
33. Jarai, G., and F. Buxton, 1994 Nitrogen, carbon, and pH regulation of extracellular acidic proteases of Aspergillus niger. Curr. Genet. 26: 238-244.
34. Jørgensen, T.R., J. Park, M. Arentshorst, A.M. van Welzen, G. Lamers et al. 2011 The molecular and genetic basis of conidial pigmentation in Aspergillus niger. Fungal Genet. Biol. 48: 544-553.
35. Kale, S.P., L. Milde, M.K. Trapp, J.C. Frisvad, N.P. Keller et al. 2008 Requirement of LaeA for secondary metabolism and sclerotial production in Aspergillus flavus. Fungal Genet. Biol. 45: 1422-1429.
36. Kamerewerd, J., I. Zadra, H. Kürnsteiner, and U. Kück, 2011 PcchiB1, encoding a class V chitinase, is affected by PcVelA and PcLaeA, and is responsible for cell wall integrity in Penicillium chrysogenum. Microbiology 157: 3036-3048.
37. Karaffa, L., and C. P. Kubicek, 2003 Aspergillus niger citric acid accumulation: do we understand this well working black box? Appl. Microbiol. Biotechnol. 61: 189-196.
38. Karimi-Aghcheh, R., J.W. Bok, P.A. Phatale, K.M. Smith, S.E. Baker et al. 2013 Functional analyses of Trichoderma reesei LAE1 reveal conserved and contrasting roles of this regulator. G3 (Bethesda) 3: 369-378.
39. Kildgaard, S., M. Mansson, I. Dosen, A. Klitgaard, J.C. Frisvad et al. 2014 Accurate dereplication of bioactive secondary metabolites from marine-derived fungi by UHPLC-DAD-QTOFMS and a MS/HRMS library. Mar. Drugs 12: 3681-3705.
40. Kosalková, K., C. García-Estrada, R.V. Ullán, R.P. Godio, R. Feltrer et al. 2009 The global regulator LaeA controls penicillin biosynthesis, pigmentation and sporulation, but not roquefortine C synthesis in Penicillium chrysogenum. Biochimie 91: 214-225.
41. Kubicek, C.P., and L. Karaffa, 2010 Bioprocess, Bioseparation and Cell Technology: Encyclopedia of Industrial Biotechnology, Ed. 7. Wiley-Blackwell, Hoboken.
42. Kubicek, C.P., G. Schreferl-Kunar, W. Wöhrer, and M. Röhr, 1988 Evidence for a cytoplasmic pathway of oxalate biosynthesis in Aspergillus niger. Appl. Environ. Microbiol. 54: 633-637.
43. Lenouvel, F., L. Fraissinet-Tachet, P.J. van de Vondervoort, and J. Visser, 2001 Isolation of UV-induced mutations in the areA nitrogen regulatory gene of Aspergillus niger, and construction of a disruption mutant. Mol. Genet. Genomics 266: 42-47.
44. Li, A., N. van Luijk, M. ter Beek, M. Caspers, P.J. Punt, M. van der Werf, 2011 A clone-based transcriptomics approach for the identification of genes relevant for itaconic acid production in Aspergillus. Fungal Genet. Biol. 48: 602-611.
45. Li, A., and P. J. Punt, 2013 Industrial production of organic acids by fungi: state of the art and opportunities, pp. 52-74 in Applications of Microbial Engineering, Vol. 1, edited by V. K. Gupta, M. Schmoll, and M. Maki. CRC Press/Taylor & Francis Group, Boca Raton.
46. Li, A., N. Pfelzer, R. Zuijderwijk, A. Brickwedde, C. van Zeijl et al. 2013 Reduced by-product formation and modified oxygen availability improve itaconic acid production in Aspergillus niger. Appl. Microbiol. Biotechnol. 97: 3901-3911.
47. MacCabe, A.P., S. Vanhanen, M.D. Sollewign Gelpke, P.J. van de Vondervoort, H.N. Arst et al. 1998 Identification, cloning and sequence of the Aspergillus niger areA wide domain regulatory gene controlling nitrogen utilisation. Biochim. Biophys. Acta 1396: 163-168.
48. Magnuson, J.K., and L. L. Lasure, 2004 Organic acid production by filamentous fungi, pp. 307-340 in Advances in Fungal Biotechnology for Industry, Agriculture, and Medicine, edited by Tkacz, J.S., and L. Lange. Kluwer Academic/Plenum Publishers, New York.
49. Mattern, I.E., J.M. van Noort, P. van den Berg, D.B. Archer, I.N. Roberts et al. 1992 Isolation and characterization of mutants of Aspergillus niger deficient in extracellular proteases. Mol. Gen. Genet. 234: 332-336.
50. Meyer, V., M. Arentshorst, S.J. Flitter, B.M. Nitsche, M.J. Kwon et al. 2009 Reconstruction of signaling networks regulating fungal morphogenesis by transcriptomics. Eukaryot. Cell 8: 1677-1691.
51. Michelmore, R.W., I. Paran, and R. V. Kesseli, 1991 Identification of markers linked to disease-resistance genes by bulked segregant analysis: a rapid method to detect markers in specific genomic regions by using segregating populations. Proc. Natl. Acad. Sci. USA 88: 9828-9832.
52. Nielsen, K.F., and T. O. Larsen, 2015 The importance of mass spectrometric dereplication in fungal secondary metabolite analysis. Front.Microbiol. 6: 71.
53. Nielsen, K.F., J.M. Mogensen, M. Johansen, T.O. Larsen, and J. C. Frisvad, 2009 Review of secondary metabolites and mycotoxins from the Aspergillus niger group. Anal. Bioanal. Chem. 395: 1225-1242.
54. Nielsen, M.L., J.B. Nielsen, C. Rank, M.L. Klejnstrup, D.K. Holm et al. 2011 A genome-wide polyketide synthase deletion library uncovers novel genetic links to polyketides and meroterpenoids in Aspergillus nidulans. FEMS Microbiol. Lett. 321: 157-166.
55. Nowrousian, M., I. Teichert, S. Masloff, and U. Kück, 2012 Whole-genome sequencing of Sordaria macrospora mutants identifies developmental genes. G3 (Bethesda) 2: 261-270.
56. Oda, K., A. Kobayashi, S. Ohashi, and M. Sano, 2011 Aspergillus oryzae laeA regulates kojic acid synthesis genes. Biosci. Biotechnol. Biochem. 75: 1832-1834.
57. Park, Y., R.M. González-Martínez, G. Navarro-Cerrillo, M. Chakroun, Y. Kim et al. 2014 ABCC transporters mediate insect resistance to multiple Bt toxins revealed by bulk segregant analysis. BMC Biol. 12: 46.
58. Pedersen, H., B. Christensen, C. Hjort, and J. Nielsen, 2000 Construction and characterization of an oxalic acid nonproducing strain of Aspergillus niger. Metab. Eng. 2: 34-41.
59. Pel, H.J., J.H. de Winde, D.B. Archer, P.S. Dyer, G. Hofmann et al. 2007 Genome sequencing and analysis of the versatile cell factory Aspergillus niger CBS 513.88. Nat. Biotechnol. 25: 221-231.
60. Peñalva, M.A., and H. N. Arst, 2002 Regulation of gene expression by ambient pH in filamentous fungi and yeasts. Microbiol. Mol. Biol. Rev. 66: 426-446.
61. Perlman, D., D.A. Kita, and W. H. Peterson, 1946 Production of citric acid from cane molasses. Arch. Biochem. 11: 123-129.
62. Perrin, R.M., N.D. Fedorova, J.W. Bok, R.A. Cramer, J.R. Wortman et al. 2007 Transcriptional regulation of chemical diversity in Aspergillus fumigatus by LaeA. PLoS Pathog. 3: e50.
63. Piumi, F., A. Levasseur, D. Navarro, S. Zhou, Y. Mathieu et al. 2014 A novel glucose dehydrogenase from the white-rot fungus Pycnoporus cinnabarinus: production in Aspergillus niger and physicochemical characterization of the recombinant enzyme. Appl. Microbiol. Biotechnol. 98: 10105-10118.
64. Pomraning, K.R., K.M. Smith, and M. Freitag, 2011 Bulk segregant analysis followed by high-throughput sequencing reveals the Neurospora cell cycle gene, ndc-1, to be allelic with the gene for ornithine decarboxylase, spe-1. Eukaryot. Cell 10: 724-733.
65. Pontecorvo, G., J.A. Roper, and E. Forbes, 1953 Genetic recombination without sexual reproduction in Aspergillus niger. J. Gen. Microbiol. 8: 198-210.
66. Poulsen, L., M.R. Andersen, A.E. Lantz, and J. Thykaer, 2012 Identification of a transcription factor controlling pH-dependent organic acid response in Aspergillus niger. PLoS One 7: e50596.
67. Punt, P.J., F.H. Schuren, J. Lehmbeck, T. Christensen, C. Hjort et al. 2008 Characterization of the Aspergillus niger prtT, a unique regulator of extracellular protease encoding genes. Fungal Genet. Biol. 45: 1591-1599.
68. Record, E., M. Asther, C. Sigoillot, S. Pagès, P.J. Punt et al. 2003 Overproduction of the Aspergillus niger feruloyl esterase for pulp bleaching application. Appl. Microbiol. Biotechnol. 62: 349-355.
69. Rose, S.H., and W. H. van Zyl, 2002 Constitutive expression of the Trichoderma reesei beta-1,4-xylanase gene (xyn2) and the beta-1,4-endoglucanase gene (egl) in Aspergillus niger in molasses and defined glucose media. Appl. Microbiol. Biotechnol. 58: 461-468.
70. Ruijter, G.J., P.J. van de Vondervoort, and J. Visser, 1999 Oxalic acid production by Aspergillus niger: an oxalate-non-producing mutant produces citric acid at pH 5 and in the presence of manganese. Microbiology 145: 2569-2576.
71. Sambrook, J., T. Maniatis, and E. F. Fritsch, 1989 Molecular Cloning: A Laboratory Manual, Ed. 2. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
72. Schachtschabel, D., M. Arentshorst, B.M. Nitsche, S. Morris, K.F. Nielsen et al. 2013 The transcriptional repressor TupA in Aspergillus niger is involved in controlling gene expression related to cell wall biosynthesis, development, and nitrogen source availability. PLoS One 8: e78102.
73. Seiboth, B., R.A. Karimi, P.A. Phatale, R. Linke, L. Hartl et al. 2012 The putative protein methyltransferase LAE1 controls cellulase gene expression in Trichoderma reesei. Mol. Microbiol. 84: 1150-1164.
74. Sugui, J.A., J. Pardo, Y.C. Chang, A. Müllbacher, K.A. Zarember et al. 2007 Role of laeA in the regulation of alb1, gliP, conidial morphology, and virulence in Aspergillus fumigatus. Eukaryot. Cell 6: 1552-1561.
75. Turbe-Doan, A., Y. Arfi, E. Record, I. Estrada-Alvarado, and A. Levasseur, 2013 Heterologous production of cellobiose dehydrogenases from the basidiomycete Coprinopsis cinerea and the ascomycete Podospora anserina and their effect on saccharification of wheat straw. Appl. Microbiol. Biotechnol. 97: 4873-4885.
76. van den Hombergh, J.P., P.J. van de Vondervoort, N.C. van der Heijden, and J. Visser, 1995 New protease mutants in Aspergillus niger result in strongly reduced in vitro degradation of target proteins; genetical and biochemical characterization of seven complementation groups. Curr. Genet. 28: 299-308.
77. van den Hombergh, J.P., A.P. MacCabe, P.J. van de Vondervoort, and J. Visser, 1996 Regulation of acid phosphatases in an Aspergillus niger pacC disruption strain. Mol. Gen. Genet. 251: 542-550.
78. van den Hombergh, J.P., M.D. Sollewijn Gelpke, P.J. van de Vondervoort, F.P. Buxton, and J. Visser, 1997a Disruption of three acid proteases in Aspergillus niger: effects on protease spectrum, intracellular proteolysis, and degradation of target proteins. Eur. J. Biochem. 247: 605-613.
79. van den Hombergh, J.P., P.J. van de Vondervoort, L. Fraissinet-Tachet, and J. Visser, 1997b Aspergillus as a host for heterologous protein production: the problem of proteases. Trends Biotechnol. 15: 256-263.
80. van de Vondervoort, P.J. I., S.M. J. Langeveld, J. Visser, N.N. M. E. van Peij, H.J. Pel et al. 2007 Identification of a mitotic recombination hotspot on chromosome III of the asexual fungus Aspergillus niger and its possible correlation with elevated basal transcription. Curr. Genet. 52: 107-114.
81. van Hartingsveldt, W., I.E. Mattern, C.M. van Zeijl, P.H. Pouwels, and C. A. van den Hondel, 1987 Development of a homologous transformation system for Aspergillus niger based on the pyrG gene. Mol. Gen. Genet. 206: 71-75.
82. Wenger, J.W., K. Schwartz, and G. Sherlock, 2010 Bulk segregant analysis by high-throughput sequencing reveals a novel xylose utilization gene from Saccharomyces cerevisiae. PLoS Genet. 6: e1000942.
83. Wiemann, P., D.W. Brown, K. Kleigrewe, J.W. Bok, N.P. Keller et al. 2010 FfVel1 and FfLae1, components of a velvet-like complex in Fusarium fujikuroi, affect differentiation, secondary metabolism and virulence. Mol. Microbiol. 77: 972-994.
84. Wu, D., S. Oide, N. Zhang, M.Y. Choi, and B. G. Turgeon, 2012 ChLae1 and ChVel1 regulate T-toxin production, virulence, oxidative stress response, and development of the maize pathogen Cochliobolus heterostrophus. PLoS Pathog. 8: e1002542.
85. Xing, W., C. Deng, and C.-H. Hu, 2010 Molecular cloning and characterization of the global regulator LaeA in Penicillium citrinum. Biotechnol. Lett. 32: 1733.
86. Zwane, E.N., S.H. Rose, W.H. van Zyl, K. Rumbold, and M. Viljoen-Bloom, 2014 Overexpression of Aspergillus tubingensis faeA in proteasedeficient Aspergillus niger enables ferulic acid production from plant material. J. Ind. Microbiol. Biotechnol. 41: 1027-1034.