Abstract
The GH10 xylanase XYNIII is expressed in the hyper-cellulase-producing mutant PC-3-7, but not in the standard strain QM9414 of Trichoderma reesei. The GH11 xylanase gene xyn1 is induced by cellulosic and xylanosic carbon sources while xyn3 is induced only by cellulosic carbon sources in the PC-3-7 strain. In this study, we constructed a modified xyn3 promoter in which we replaced the cis-acting region of the xyn3 promoter by the cis-acting region of the xyn1 promoter. The resulting xyn3 chimeric promoter exhibited improved inductivity against cellulosic carbon over the wild-type promoter and acquired inductivity against xylanosic carbon. Furthermore, PC-3-7 expressing the heterologous β-glycosidase gene, Aspergillus aculeatus bgl1, under the control of the xyn3 chimeric promoter, showed enhanced saccharification ability through increased cellobiase activity. We also show that the xyn3 chimeric promoter is also functional in the QM9414 strain. Our results indicate that the xyn3 chimeric promoter is very efficient for enzyme expression.
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Aro N, Saloheimo A, Ilmén M, Penttilä M (2001) ACEII, a novel transcriptional activator involved in regulation of cellulase and xylanase genes of Trichoderma reesei. J Biol Chem 276(26):24309–24314. https://doi.org/10.1074/jbc.M003624200
Aro N, Ilmén M, Saloheimo A, Penttilä M (2003) ACEI of Trichoderma reesei is a repressor of cellulase and xylanase expression. Appl Environ Microbiol 69(1):56–65. https://doi.org/10.1128/AEM.69.1.56-65.2003
Derntl C, Rassinger A, Srebotnik E, Mach RL, Mach-Aigner AR (2015) Xpp1 regulates the expression of xylanases, but not of cellulases in Trichoderma reesei. Biotechnol Biofuels 8(1):112. https://doi.org/10.1186/s13068-015-0298-8
Foreman PK, Brown D, Dankmeyer L, Dean R, Diener S, Dunn-Coleman NS, Goedegebuur F, Houfek TD, England GJ, Kelley AS, Meerman HJ, Mitchell T, Mitchinson C, Olivares HA, Teunissen PJM, Yao J, Ward M (2003) Transcriptional regulation of biomass-degrading enzymes in the filamentous fungus Trichoderma reesei. J Biol Chem 278(34):31988–31997. https://doi.org/10.1074/jbc.M304750200
Fowler T, Brown RD (1992) The bgl1 gene encoding extracellular ß-glucosidase from Trichoderma reesei is required for rapid induction of the cellulase complex. Mol Microbiol 6(21):3225–3235. https://doi.org/10.1111/j.1365-2958.1992.tb01777.x
Furukawa T, Shida Y, Kitagami N, Ota Y, Adachi M, Nakagawa S, Shimada R, Kato M, Kobayashi T, Okada H, Ogasawara W, Morikawa Y (2008) Identification of the cis-acting elements involved in regulation of xylanase III gene expression in Trichoderma reesei PC-3-7. Fungal Genet Biol 45(7):1094–1102. https://doi.org/10.1016/j.fgb.2008.03.006
Furukawa T, Shida Y, Kitagami N, Mori K, Kato M, Kobayashi T, Okada H, Ogasawara W, Morikawa Y (2009) Identification of specific binding sites for XYR1, a transcriptional activator of cellulolytic and xylanolytic genes in Trichoderma reesei. Fungal Genet Biol 46(8):564–574. https://doi.org/10.1016/j.fgb.2009.04.001
Häkkinen M, Valkonen MJ, Westerholm-Parvinen A, Aro N, Arvas M, Vitikainen M, Penttilä M, Saloheimo M, Pakula TM (2014) Screening of candidate regulators for cellulase and hemicellulase production in Trichoderma reesei and identification of a factor essential for cellulase production. Biotechnol Biofuels 7(1):14. https://doi.org/10.1186/1754-6834-7-14
Herold S, Bischof R, Metz B, Seiboth B, Kubicek CP (2013) Xylanase gene transcription in Trichoderma reesei is triggered by different inducers representing different hemicellulosic pentose polymers. Eukaryot Cell 12(3):390–398. https://doi.org/10.1128/EC.00182-12
Herrmann MC, Vrsanska M, Jurickova M, Hirsch J, Biely P, Kubicek CP (1997) The ß-D-xylosidase of Trichoderma reesei is a multifunctional ß-D-xylan xylohydrolase. Biochem J 321(2):375–381. https://doi.org/10.1042/bj3210375
Hynes MJ, Corrick CM, King JA (1983) Isolation of genomic clones containing the amdS gene of Aspergillus nidulans and their use in the analysis of structural and regulatory mutations. Mol Cell Biol 3(8):1430–1439. https://doi.org/10.1128/MCB.3.8.1430.Updated
Ilmén M, Saloheimo A, Onnela ML, Penttilä M (1997) Regulation of cellulase gene expression in the filamentous fungus Trichoderma reesei. Appl Environ Microbiol 63(4):1298–1306
Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Report 5(4):387–405. https://doi.org/10.1007/BF02667740
Karlsson J, Saloheimo M, Siika-Aho M, Tenkanen M, Penttilä M, Tjerneld F (2001) Homologous expression and characterization of Cel61A (EG IV) of Trichoderma reesei. Eur J Biochem 268(24):6498–6507. https://doi.org/10.1046/j.0014-2956.2001.02605.x
Kawai T, Nakazawa H, Ida N, Okada H, Tani S, Sumitani J-I, Kawaguchi T, Ogasawara W, Morikawa Y, Kobayashi Y (2012) Analysis of the saccharification capability of high-functional cellulase JN11 for various pretreated biomasses through a comparison with commercially available counterparts. J Ind Microbiol Biotechnol 39(12):1741–1749. https://doi.org/10.1007/s10295-012-1195-9
Kawamori M, Morikawa Y, Takasawa S (1986) Induction and production of cellulases by L-sorbose in Trichoderma reesei. Appl Microbiol Biotechnol 24(6):449–453. https://doi.org/10.1007/BF00250321
Liu T, Wang T, Li X, Liu X (2008) Improved heterologous gene expression in Trichoderma reesei by cellobiohydrolase I gene (cbh1) promoter optimization. Acta Biochim Biophys Sin Shanghai 40(2):158–165. https://doi.org/10.1111/j.1745-7270.2008.00388.x
Liu R, Chen L, Jiang Y, Zou G, Zhou Z (2017) A novel transcription factor specifically regulates GH11 xylanase genes in Trichoderma reesei. Biotechnol Biofuels 10(1):194. https://doi.org/10.1186/s13068-017-0878-x
Mach RL, Strauss J, Zeilinger S, Schindler M, Kubicek CP (1996) Carbon catabolite repression of xylanase I (xyn1) gene expression in Trichoderma reesei. Mol Microbiol 21(6):1273–1281. https://doi.org/10.1046/j.1365-2958.1996.00094.x
Mach-Aigner AR, Grosstessner-Hain K, Poças-Fonseca MJ, Mechtler K, Mach RL (2010) From an electrophoretic mobility shift assay to isolated transcription factors: a fast genomic-proteomic approach. BMC Genomics 11(1):644. https://doi.org/10.1186/1471-2164-11-644
Maruyama JI, Kitamoto K (2008) Multiple gene disruptions by marker recycling with highly efficient gene-targeting background (∆ligD) in Aspergillus oryzae. Biotechnol Lett 30(10):1811–1817. https://doi.org/10.1007/s10529-008-9763-9
Minetoki T, Kumagai C, Gomi K, Kitamoto K, Takahashi K (1998) Improvement of promoter activity by the introduction of multiple copies of the conserved region III sequence, involved in the efficient expression of Aspergillus oryzae amylase-encoding genes. Appl Microbiol Biotechnol 50(4):459–467. https://doi.org/10.1007/s002530051321
Nakazawa H, Kawai T, Ida N, Shida Y, Kobayashi Y, Okada H, Tani S, Sumitani J-I, Kawaguchi T, Morikawa Y, Ogasawara W (2012) Construction of a recombinant Trichoderma reesei strain expressing Aspergillus aculeatus β-glucosidase 1 for efficient biomass conversion. Biotechnol Bioeng 109(1):92–99. https://doi.org/10.1002/bit.23296
Nakazawa H, Kawai T, Ida N, Shida Y, Shioya K, Kobayashi Y, Okada H, Tani S, Sumitani J-I, Kawaguchi T, Morikawa Y, Ogasawara W (2016) A high performance Trichoderma reesei strain that reveals the importance of xylanase III in cellulosic biomass conversion. Enzym Microb Technol 82:89–95. https://doi.org/10.1016/j.enzmictec.2015.08.019
Nitta M, Furukawa T, Shida Y, Mori K, Kuhara S, Morikawa Y, Ogasawara W (2012) A new Zn(II)2Cys6-type transcription factor BglR regulates ß-glucosidase expression in Trichoderma reesei. Fungal Genet Biol 49(5):388–397. https://doi.org/10.1016/j.fgb.2012.02.009
Nogawa M, Goto M, Okada H, Morikawa Y (2001) L-sorbose induces cellulase gene transcription in the cellulolytic fungus Trichoderma reesei. Curr Genet 38(6):329–334. https://doi.org/10.1007/s002940000165
Ogasawara W, Shida Y, Furukawa T, Shimada R, Nakagawa S, Kawamura M, Yagyu T, Kosuge A, Xu J, Nogawa M, Okada H, Morikawa Y (2006) Cloning, functional expression and promoter analysis of xylanase III gene from Trichoderma reesei. Appl Microbiol Biotechnol 72(5):995–1003. https://doi.org/10.1007/s00253-006-0365-y
Okada H, Tada K, Sekiya T, Yokoyama K, Takahashi A, Tohda H, Kumagai H, Morikawa Y (1998) Molecular characterization and heterologous expression of the gene encoding a low-molecular-mass endoglucanase from Trichoderma reesei QM9414. Appl Environ Microbiol 64(2):555–563
Penttilä M, Lehtovaara P, Nevalainen H, Bhikhabhai R, Knowles J (1986) Homology between cellulase genes of Trichoderma reesei: complete nucleotide sequence of the endoglucanase I gene. Gene 45(3):253–263. https://doi.org/10.1016/0378-1119(86)90023-5
Penttilä M, Nevalainen H, Rättö M, Salminen E, Knowles J (1987) A versatile transformation system for the cellulolytic filamentous fungus Trichoderma reesei. Gene 61(2):155–164. https://doi.org/10.1016/0378-1119(87)90110-7
Porciuncula JDO, Furukawa T, Mori K, Shida Y, Hirakawa H, Tashiro K, Kuhara S, Nakagawa S, Morikawa Y, Ogasawara W (2013) Single nucleotide polymorphism analysis of a Trichoderma reesei hyper-cellulolytic mutant developed in Japan. Biosci Biotechnol Biochem 77(3):534–543. https://doi.org/10.1271/bbb.120794
Portnoy T, Margeot A, Linke R, Atanasova L, Fekete E, Sandor E, Hartl L, Karaffa L, Druzhinina IS, Seiboth B, Le Crom S, Kubicek CP, Sándor E, Hartl L, Karaffa L, Druzhinina IS, Seiboth B, Le Crom S, Kubicek CP (2011) The CRE1 carbon catabolite repressor of the fungus Trichoderma reesei: a master regulator of carbon assimilation. BMC Genomics 12(1):269. https://doi.org/10.1186/1471-2164-12-269
Rahman Z, Shida Y, Furukawa T, Suzuki Y, Okada H, Ogasawara W, Morikawa Y (2009a) Evaluation and characterization of Trichoderma reesei cellulase and xylanase promoters. Appl Microbiol Biotechnol 82(5):899–908. https://doi.org/10.1007/s00253-008-1841-3
Rahman Z, Shida Y, Furukawa T, Suzuki Y, Okada H, Ogasawara W, Morikawa Y (2009b) Application of Trichoderma reesei cellulase and xylanase promoters through homologous recombination for enhanced production of extracellular β-glucosidase I. Biosci Biotechnol Biochem 73(5):1083–1089. https://doi.org/10.1271/bbb.80852
Rauscher R, Würleitner E, Wacenovsky C, Aro N, Stricker AR, Zeilinger S, Kubicek CP, Penttilä M, Mach RL (2006) Transcriptional regulation of xyn1, encoding xylanase I, in Hypocrea jecorina. Eukaryot Cell 5(3):447–456. https://doi.org/10.1128/EC.5.3.447-456.2006
Saloheimo M, Lehtovaara P, Penttilä M, Teeri TT, Ståhlberg J, Johansson G, Pettersson G, Claeyssens M, Tomme P, Knowles JKC (1988) EGIII, a new endoglucanase from Trichoderma reesei: the characterization of both gene and enzyme. Gene 63(1):11–21. https://doi.org/10.1016/0378-1119(88)90541-0
Saloheimo A, Henrissat B, Hoffrén AM, Teleman O, Penttilä M (1994) A novel, small endoglucanase gene, egl5, from Trichoderma reesei isolated by expression in yeast. Mol Microbiol 13(2):219–228. https://doi.org/10.1111/j.1365-2958.1994.tb00417.x
Saloheimo M, Nakari-Setälä T, Tenkanen M, Penttilä M (1997) cDNA cloning of a Trichoderma reesei cellulase and demonstration of endoglucanase activity by expression in yeast. Eur J Biochem 249(2):584–591. https://doi.org/10.1111/j.1432-1033.1997.00584.x
Saloheimo A, Aro N, Ilmén M, Penttilä M (2000) Isolation of the ace1 gene encoding a Cys2-His2 transcription factor involved in regulation of activity of the cellulase promoter cbh1 of Trichoderma reesei. J Biol Chem 275(8):5817–5825. https://doi.org/10.1074/jbc.275.8.5817
Saloheimo M, Kuja-Panula J, Ylösmäki E, Ward M, Penttilä M (2002) Enzymatic properties and intracellular localization of the novel Trichoderma reesei β-glucosidase BGLII (Cel1A). Appl Environ Microbiol 68(9):4546–4553. https://doi.org/10.1128/AEM.68.9.4546-4553.2002
Schmoll M, Schuster A (2010) Biology and biotechnology of Trichoderma. Appl Microbiol Biotechnol 87(3):787–799. https://doi.org/10.1007/s00253-010-2632-1
Seiboth B, Hartl L, Pail M, Fekete E, Karaffa L, Kubicek CP (2004) The galactokinase of Hypocrea jecorina is essential for cellulose induction by lactose but dispensable for growth on D-galactose. Mol Microbiol 51(4):1015–1025. https://doi.org/10.1046/j.1365-2958.2003.03901.x
Shida Y, Yamaguchi K, Nitta M, Nakamura A, Takahashi M, Kidokoro S, Mori K, Tashiro K, Kuhara S, Matsuzawa T, Yaoi K, Sakamoto Y, Tanaka N, Morikawa Y, Ogasawara W (2015) The impact of a single-nucleotide mutation of bgl2 on cellulase induction in a Trichoderma reesei mutant. Biotechnol Biofuels 8(1):230. https://doi.org/10.1186/s13068-015-0420-y
Shoemaker S, Schweickart V, Ladner M, Gelfand D, Kwok S, Myambo K, Innis M (1983) Molecular cloning of exo–cellobiohydrolase I derived from Trichoderma reesei strain L27. Nat Biotechnol 1(8):691–696. https://doi.org/10.1038/nbt1083-691
Sluiter A, Hames B, Ruiz R, Scarlata C, Sluiter J, Templeton D, Nrel DC (2008) Determination of structural carbohydrates and lignin in biomass. NREL laboratory analytical procedures. http://www.eere.energy.gov/ biomass/analytical_procedures.html#LAP-002. Accessed 17 July 2005
Strauss J, Mach RL, Zeilinger S, Hartler G, Stöffler G, Wolschek M, Kubicek CP (1995) Cre1, the carbon catabolite repressor protein from Trichoderma reesei. FEBS Lett 376(1-2):103–107. https://doi.org/10.1016/0014-5793(95)01255-5
Stricker AR, Grosstessner-Hain K, Würleitner E, Mach RL (2006) Xyr1 (Xylanase Regulator 1) regulates both the hydrolytic enzyme system and D-xylose metabolism in Hypocrea jecorina. Eukaryot Cell 5(12):2128–2137. https://doi.org/10.1128/EC.00211-06
Stricker AR, Steiger MG, Mach RL (2007) Xyr1 receives the lactose induction signal and regulates lactose metabolism in Hypocrea jecorina. FEBS Lett 581(21):3915–3920. https://doi.org/10.1016/j.febslet.2007.07.025
Stricker AR, Trefflinger P, Aro N, Penttilä M, Mach RL (2008) Role of Ace2 (Activator of Cellulases 2) within the xyn2 transcriptosome of Hypocrea jecorina. Fungal Genet Biol 45(4):436–445. https://doi.org/10.1016/j.fgb.2007.08.005
Suto M, Tomita F (2001) Induction and catabolite repression mechanisms of cellulase in fungi. J Biosci Bioeng 92(4):305–311. https://doi.org/10.1016/S1389-1723(01)80231-0
Takada G, Kawaguchi T, Sumitani J, Arai M (1998) Expression of Aspergillus aculeatus no. F-50 cellobiohydrolase I (cbhI) and ß-glucosidase 1 (bgl1) genes by Saccharomyces cerevisiae. Biosci Biotechnol Biochem 62(8):1615–1618. https://doi.org/10.1271/bbb.62.1615
Teeri TT, Lehtovaara P, Kauppinen S, Salovuori I, Knowles J (1987) Homologous domains in Trichoderma reesei cellulolytic enzymes: gene sequence and expression of cellobiohydrolase II. Gene 51(1):43–52. https://doi.org/10.1016/0378-1119(87)90472-0
Tenkanen M, Puls J, Poutanen K (1992) Two major xylanases of Trichoderma reesei. Enzym Microb Technol 14(7):566–574. https://doi.org/10.1016/0141-0229(92)90128-B
Tenkanen M, Vršanská M, Siika-Aho M, Wong DW, Puchart V, Penttilä M, Saloheimo M, Biely P (2013) Xylanase XYN IV from Trichoderma reesei showing exo- and endo-xylanase activity. FEBS J 280(1):285–301. https://doi.org/10.1111/febs.12069
Xu J, Takakuwa N, Nogawa M, Okada H, Morikawa Y (1998) A third xylanase from Trichoderma reesei PC-3-7. Appl Microbiol Biotechnol 49(6):718–724. https://doi.org/10.1007/s002530051237
Xu J, Nogawa M, Okada H, Morikawa Y (2000) Regulation of xyn3 gene expression in Trichoderma reesei PC-3-7. Appl Microbiol Biotechnol 54(3):370–375. https://doi.org/10.1007/s002530000410
Zeilinger S, Mach RL, Schindler M, Herzog P, Kubicek CP (1996) Different inducibility of expression of the two xylanase genes xyn1 and xyn2 in Trichoderma reesei. J Biol Chem 271(41):25624–25629. https://doi.org/10.1074/jbc.271.41.25624
Acknowledgements
The authors thank Dr. Hiroshi Kakeshita from Kao Co., Ltd. for providing the anti AaBGLI antibody and Dr. Yoshinori Kobayashi from Japan Bioindustry Association for providing the pretreated erianthus and eucalyptus.
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This work was supported by a grant from the New Energy and Industrial Technology Development Organization (NEDO) Project (P07015 and P13011).
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Hirasawa, H., Shioya, K., Furukawa, T. et al. Engineering of the Trichoderma reesei xylanase3 promoter for efficient enzyme expression. Appl Microbiol Biotechnol 102, 2737–2752 (2018). https://doi.org/10.1007/s00253-018-8763-5
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DOI: https://doi.org/10.1007/s00253-018-8763-5