Abstract
Previously identified twelve plant cell wall degradation-associated genes of the white rot fungus Phlebia radiata were studied by RT-qPCR in semi-aerobic solid-state cultures on lignocellulose waste material, and on glucose-containing reference medium. Wood-decay-involved enzyme activities and ethanol production were followed to elucidate both the degradative and fermentative processes. On the waste lignocellulose substrate, P. radiata carbohydrate-active enzyme (CAZy) genes encoding cellulolytic and hemicellulolytic activities were significantly upregulated whereas genes involved in lignin modification displayed a more complex response. Two lignin peroxidase genes were differentially expressed on waste lignocellulose compared to glucose medium, whereas three manganese peroxidase-encoding genes were less affected. On the contrary, highly significant difference was noticed for three cellulolytic genes (cbhI_1, eg1, bgl1) with higher expression levels on the lignocellulose substrate than on glucose. This indicates expression of the wood-attacking degradative enzyme system by the fungus also on the recycled, waste core board material. During the second week of cultivation, ethanol production increased on the core board to 0.24 g/L, and extracellular activities against cellulose, xylan, and lignin were detected. Sugar release from the solid lignocellulose resulted with concomitant accumulation of ethanol as fermentation product. Our findings confirm that the fungus activates its white rot decay system also on industrially processed lignocellulose adopted as growth substrate, and under semi-aerobic cultivation conditions. Thus, P. radiata is a good candidate for lignocellulose-based renewable biotechnology to make biofuels and biocompounds from materials with less value for recycling or manufacturing.
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References
Agger JW, Isaksen T, Várnai A, Vidal-Melgosa S, Willats WGT, Ludwig R, Horn SJ, Eijsink VGH, Westereng B (2014) Discovery of LPMO activity on hemicelluloses shows the importance of oxidative processes in plant cell wall degradation. Proc Natl Acad Sci U S A 111:6287–6292. https://doi.org/10.1073/pnas.1323629111
Andersen CL, Jensen JL, Ørntoft TF (2004) Normalization of real-time quantitative reverse transcription-pcr data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res 64:5245–5250. https://doi.org/10.1158/0008-5472.CAN-04-0496
Beeson WT, Vu VV, Span EA, Phillips CM, Marletta MA (2015) Cellulose degradation by polysaccharide monooxygenases. Annu Rev Biochem 84:923–946. https://doi.org/10.1146/annurev-biochem-060614-034439
Boerjan W, Ralph J, Baucher M (2003) Lignin biosynthesis. Annu Rev Plant Biol 54:519–546. https://doi.org/10.1146/annurev.arplant.54.031902.134938
Broda P, Birch PR, Brooks PR, Sims PF (1995) PCR-mediated analysis of lignocellulolytic gene transcription by Phanerochaete chrysosporium: substrate-dependent differential expression within gene families. Appl Environ Microbiol 61:2358–2364
Castanera R, López-Varas L, Pisabarro AG, Ramírez L (2015) Validation of reference genes for transcriptional analyses in Pleurotus ostreatus by using reverse transcription-quantitative PCR. Appl Environ Microbiol 81:4120–4129. https://doi.org/10.1128/AEM.00402-15
Cohen R, Yarden O, Hadar Y (2002) Lignocellulose affects Mn2+ regulation of peroxidase transcript levels in solid-state cultures of Pleurotus ostreatus. Appl Environ Microbiol 68:3156–3158. https://doi.org/10.1128/AEM.68.6.3156-3158.2002
Courtade G, Wimmer R, Røhr ÅK, Preims M, Felice AKG, Dimarogona M, Vaaje-Kolstad G, Sørlie M, Sandgren M, Ludwig R, Eijsink VGH, Aachmann FL (2016) Interactions of a fungal lytic polysaccharide monooxygenase with β-glucan substrates and cellobiose dehydrogenase. Proc Natl Acad Sci U S A 113:5922–5927. https://doi.org/10.1073/pnas.1602566113
Eastwood DC, Floudas D, Binder M, Majcherczyk A, Schneider P, Aerts A, Asiegbu FO, Baker SE, Barry K, Bendiksby M, Blumentritt M, Coutinho PM, Cullen D, de Vries RP, Gathman A, Goodell B, Henrissat B, Ihrmark K, Kauserud H, Kohler A, LaButti K, Lapidus A, Lavin JL, Lee Y-H, Lindquist E, Lilly W, Lucas S, Morin E, Murat C, Oguiza JA, Park J, Pisabarro AG, Riley R, Rosling A, Salamov A, Schmidt O, Schmutz J, Skrede I, Stenlid J, Wiebenga A, Xie X, Kües U, Hibbett DS, Hoffmeister D, Högberg N, Martin F, Grigoriev IV, Watkinson SC (2011) The plant cell wall–decomposing machinery underlies the functional diversity of forest fungi. Science 333:762–765. https://doi.org/10.1126/science.1205411
Floudas D, Binder M, Riley R, Barry K, Blanchette RA, Henrissat B, Martinez AT, Otillar R, Spatafora JW, Yadav JS, Aerts A, Benoit I, Boyd A, Carlson A, Copeland A, Coutinho PM, de Vries RP, Ferreira P, Findley K, Foster B, Gaskell J, Glotzer D, Gorecki P, Heitman J, Hesse C, Hori C, Igarashi K, Jurgens JA, Kallen N, Kersten P, Kohler A, Kües U, Kumar TKA, Kuo A, LaButti K, Larrondo LF, Lindquist E, Ling A, Lombard V, Lucas S, Lundell T, Martin R, McLaughlin DJ, Morgenstern I, Morin E, Murat C, Nagy LG, Nolan M, Ohm RA, Patyshakuliyeva A, Rokas A, Ruiz-Duenas FJ, Sabat G, Salamov A, Samejima M, Schmutz J, Slot JC, St. John F, Stenlid J, Sun H, Sun S, Syed K, Tsang A, Wiebenga A, Young D, Pisabarro A, Eastwood DC, Martin F, Cullen D, Grigoriev IV, Hibbett DS (2012) The paleozoic origin of enzymatic lignin decomposition reconstructed from 31 fungal genomes. Science 336:1715–1719. https://doi.org/10.1126/science.1221748
Gettemy JM, Ma B, Alic M, Gold MH (1998) Reverse transcription-PCR analysis of the regulation of the manganese peroxidase gene family. Appl Environ Microbiol 64:569–574
Gillet S, Aguedo M, Petitjean L, Morais ARC, da Costa Lopes AM, Łukasik RM, Anastas PT (2017) Lignin transformations for high value applications: towards targeted modifications using green chemistry. Green Chem 19:4200–4233. https://doi.org/10.1039/C7GC01479A
Hakala TK, Maijala P, Konn J, Hatakka A (2004) Evaluation of novel wood-rotting polypores and corticioid fungi for the decay and biopulping of Norway spruce (Picea abies) wood. Enzym Microb Technol 34:255–263. https://doi.org/10.1016/j.enzmictec.2003.10.014
Hammel K, Cullen D (2008) Role of fungal peroxidases in biological ligninolysis. Curr Opin Plant Biol 11:349–355. https://doi.org/10.1016/j.pbi.2008.02.003
Hatakka A, Hammel KE (2011) Fungal biodegradation of lignocellulose. In: Hofrichter M (ed) The Mycota, industrial applications, second edn. Springer-Verlag, Berlin Heidelberg, pp 319–340
Hemsworth GR, Johnston EM, Davies GJ, Walton PH (2015) Lytic polysaccharide monooxygenases in biomass conversion. Trends Biotechnol 33:747–761. https://doi.org/10.1016/j.tibtech.2015.09.006
Hildén K, Martinez AT, Hatakka A, Lundell T (2005) The two manganese peroxidases Pr-MnP2 and Pr-MnP3 of Phlebia radiata, a lignin-degrading basidiomycete, are phylogenetically and structurally divergent. Fungal Genet Biol 42:403–419. https://doi.org/10.1016/j.fgb.2005.01.008
Hildén KS, Mäkelä MR, Hakala TK, Hatakka A, Lundell T (2006) Expression on wood, molecular cloning and characterization of three lignin peroxidase (LiP) encoding genes of the white rot fungus Phlebia radiata. Curr Genet 49:97–105. https://doi.org/10.1007/s00294-005-0045-y
Hofrichter M, Lundell T, Hatakka A (2001) Conversion of milled pine wood by manganese peroxidase from Phlebia radiata. Appl Environ Microbiol 67:4588–4593. https://doi.org/10.1128/AEM.67.10.4588-4593.2001
Hofrichter M, Ullrich R, Pecyna MJ, Liers C, Lundell T (2010) New and classic families of secreted fungal heme peroxidases. Appl Microbiol Biotechnol 87:871–897. https://doi.org/10.1007/s00253-010-2633-0
Hori C, Ishida T, Igarashi K, Samejima M, Suzuki H, Master E, Ferreira P, Ruiz-Dueñas FJ, Held B, Canessa P, Larrondo LF, Schmoll M, Druzhinina IS, Kubicek CP, Gaskell JA, Kersten P, St John F, Glasner J, Sabat G, Splinter BonDurant S, Syed K, Yadav J, Mgbeahuruike AC, Kovalchuk A, Asiegbu FO, Lackner G, Hoffmeister D, Rencoret J, Gutiérrez A, Sun H, Lindquist E, Barry K, Riley R, Grigoriev IV, Henrissat B, Kües U, Berka RM, Martínez AT, Covert SF, Blanchette RA, Cullen D (2014) Analysis of the Phlebiopsis gigantea genome, transcriptome and secretome provides insight into its pioneer colonization strategies of wood. PLoS Genet 10:e1004759. https://doi.org/10.1371/journal.pgen.1004759
Horn S, Vaaje-Kolstad G, Westereng B, Eijsink VG (2012) Novel enzymes for the degradation of cellulose. Biotechnol Biofuels 5:45. https://doi.org/10.1186/1754-6834-5-45
Janse BJH, Gaskell J, Akhtar M, Cullen D (1998) Expression of Phanerochaete chrysosporium genes encoding lignin peroxidases, manganese peroxidases, and glyoxal oxidase in wood. Appl Environ Microbiol 64:3536–3538
Johansson T, Nyman PO, Cullen D (2002) Differential regulation of mnp2, a new manganese peroxidase-encoding gene from the ligninolytic fungus Trametes versicolor PRL 572. Appl Environ Microbiol 68:2077–2080. https://doi.org/10.1128/AEM.68.4.2077-2080.2002
Kamei I, Daikoku C, Tsutsumi Y, Kondo R (2008) Saline-dependent regulation of manganese peroxidase genes in the hypersaline-tolerant white rot fungus Phlebia sp. strain MG-60. Appl Environ Microbiol 74:2709–2716. https://doi.org/10.1128/AEM.02257-07
Kamei I, Hirota Y, Meguro S (2012a) Integrated delignification and simultaneous saccharification and fermentation of hard wood by a white-rot fungus, Phlebia sp. MG-60. Bioresour Technol 126:137–141. https://doi.org/10.1016/j.biortech.2012.09.007
Kamei I, Hirota Y, Mori T, Hirai H, Meguro S, Kondo R (2012b) Direct ethanol production from cellulosic materials by the hypersaline-tolerant white-rot fungus Phlebia sp.MG-60. Bioresour Technol 112:137–142. https://doi.org/10.1016/j.biortech.2012.02.109
Korripally P, Hunt CG, Houtman CJ, Jones DC, Kitin PJ, Cullen D, Hammel KE (2015) Regulation of gene expression during the onset of ligninolytic oxidation by Phanerochaete chrysosporium on spruce wood. Appl Environ Microbiol 81:7802–7812. https://doi.org/10.1128/AEM.02064-15
Kuuskeri J, Mäkelä MR, Isotalo J, Oksanen I, Lundell T (2015) Lignocellulose-converting enzyme activity profiles correlate with molecular systematics and phylogeny grouping in the incoherent genus Phlebia (Polyporales, Basidiomycota). BMC Microbiol 15:217. https://doi.org/10.1186/s12866-015-0538-x
Kuuskeri J, Häkkinen M, Laine P, Smolander O-P, Tamene F, Miettinen S, Nousiainen P, Kemell M, Auvinen P, Lundell T (2016) Time-scale dynamics of proteome and transcriptome of the white-rot fungus Phlebia radiata: growth on spruce wood and decay effect on lignocellulose. Biotechnol Biofuels 9:192. https://doi.org/10.1186/s13068-016-0608-9
Langston JA, Shaghasi T, Abbate E, Xu F, Vlasenko E, Sweeney MD (2011) Oxidoreductive cellulose depolymerization by the enzymes cellobiose dehydrogenase and glycoside hydrolase 61. Appl Environ Microbiol 77:7007–7015. https://doi.org/10.1128/AEM.05815-11
Linke D, Lehnert N, Nimtz M, Berger RG (2014) An alcohol oxidase of Phanerochaete chrysosporium with a distinct glycerol oxidase activity. Enzym Microb Technol 61–62:7–12. https://doi.org/10.1016/j.enzmictec.2014.04.001
Llanos A, François J, Parrou J-L (2015) Tracking the best reference genes for RT-qPCR data normalization in filamentous fungi. BMC Genomics 16:71. https://doi.org/10.1186/s12864-015-1224-y
Lombard V, Golaconda Ramulu H, Drula E, Coutinho PM, Henrissat B (2014) The carbohydrate-active enzymes database (CAZy) in 2013. Nucleic Acids Res 42:D490–D495. https://doi.org/10.1093/nar/gkt1178
Lundell T, Wever R, Floris R, Harvey P, Hatakka A, Brunow G, Schoemaker H (1993) Lignin peroxidase L3 from Phlebia radiata. Pre-steady-state and steady-state studies with veratryl alcohol and a non-phenolic lignin model compound 1-(3,4-dimethoxyphenyl)-2-(2-methoxyphenoxy)propane-1,3-diol. Eur J Biochem 211:391–4020. https://doi.org/10.1111/j.1432-1033.1993.tb17562.x
Lundell TK, Mäkelä MR, Hildén K (2010) Lignin-modifying enzymes in filamentous basidiomycetes--ecological, functional and phylogenetic review. J Basic Microbiol 50:5–20. https://doi.org/10.1002/jobm.200900338
Lundell TK, Mäkelä MR, de Vries RP, Hildén KS (2014) Genomics, lifestyles and future prospects of wood-decay and litter-decomposing Basidiomycota. In: Martin, F (ed) Fungi. Adv Bot Res 70: 329–370. Academic Press, Oxford: United Kingdom. doi: https://doi.org/10.1016/B978-0-12-397940-7.00011-2
Lundell T, Bentley E, Hilden K, Rytioja J, Kuuskeri J, Ufot UF, Nousiainen P, Hofrichter M, Wahlsten M, Doyle W, Smith AT (2017) Engineering towards catalytic use of fungal class-II peroxidases for dye-decolorizing and conversion of lignin model compounds. Curr Biotechnol 6:116–127. https://doi.org/10.2174/2211550105666160520120101
Macdonald J, Master ER (2012) Time-dependent profiles of transcripts encoding lignocellulose-modifying enzymes of the white rot fungus Phanerochaete carnosa grown on multiple wood substrates. Appl Environ Microbiol 78:1596–1600. https://doi.org/10.1128/AEM.06511-11
MacDonald J, Doering M, Canam T, Gong Y, Guttman DS, Campbell MM, Master ER (2011) Transcriptomic responses of the softwood-degrading white-rot fungus Phanerochaete carnosa during growth on coniferous and deciduous wood. Appl Environ Microbiol 77:3211–3218. https://doi.org/10.1128/AEM.02490-10
MacDonald J, Suzuki H, Master ER (2012) Expression and regulation of genes encoding lignocellulose-degrading activity in the genus Phanerochaete. Appl Microbiol Biotechnol 94:339–351. https://doi.org/10.1007/s00253-012-3937-z
Mäkelä MR, Hildén KS, Hakala TK, Hatakka A, Lundell TK (2006) Expression and molecular properties of a new laccase of the white rot fungus Phlebia radiata grown on wood. Curr Genet 50:323–333. https://doi.org/10.1007/s00294-006-0090-1
Mali T, Kuuskeri J, Shah F, Lundell TK (2017) Interactions affect hyphal growth and enzyme profiles in combinations of coniferous wood-decaying fungi of Agaricomycetes. PLoS One 12:e0185171. https://doi.org/10.1371/journal.pone.0185171
Mancilla RA, Canessa P, Manubens A, Vicuña R (2010) Effect of manganese on the secretion of manganese-peroxidase by the basidiomycete Ceriporiopsis subvermispora. Fungal Genet Biol 47:656–661. https://doi.org/10.1016/j.fgb.2010.04.003
Marinović M, Aguilar-Pontes MV, Zhou M, Miettinen O, de Vries RP, Mäkelä MR, Hildén K (2017) Temporal transcriptome analysis of the white-rot fungus Obba rivulosa shows expression of a constitutive set of plant cell wall degradation targeted genes during growth on solid spruce wood. Fungal Genet Biol 112:47–54. https://doi.org/10.1016/j.fgb.2017.07.004
Marshall OJ (2004) PerlPrimer: cross-platform, graphical primer design for standard, bisulphite and real-time PCR. Bioinformatics 20:2471–2472. https://doi.org/10.1093/bioinformatics/bth254
Martínez ÁT, Ruiz-Dueñas FJ, Martínez MJ, del Río JC, Gutiérrez A (2009) Enzymatic delignification of plant cell wall: from nature to mill. Curr Opin Biotechnol 20:348–357. https://doi.org/10.1016/j.copbio.2009.05.002
Martinez D, Challacombe J, Morgenstern I, Hibbett D, Schmoll M, Kubicek CP, Ferreira P, Ruiz-Duenas FJ, Martinez AT, Kersten P, Hammel KE, Vanden Wymelenberg A, Gaskell J, Lindquist E, Sabat G, Bondurant SS, Larrondo LF, Canessa P, Vicuna R, Yadav J, Doddapaneni H, Subramanian V, Pisabarro AG, Lavín JL, Oguiza JA, Master E, Henrissat B, Coutinho PM, Harris P, Magnuson JK, Baker SE, Bruno K, Kenealy W, Hoegger PJ, Kües U, Ramaiya P, Lucas S, Salamov A, Shapiro H, Tu H, Chee CL, Misra M, Xie G, Teter S, Yaver D, James T, Mokrejs M, Pospisek M, Grigoriev IV, Brettin T, Rokhsar D, Berka R, Cullen D (2009) Genome, transcriptome, and secretome analysis of wood decay fungus Postia placenta supports unique mechanisms of lignocellulose conversion. Proc Natl Acad Sci U S A 106:1954–1959. https://doi.org/10.1073/pnas.0809575106
Mattila H, Kuuskeri J, Lundell T (2017) Single-step, single-organism bioethanol production and bioconversion of lignocellulose waste materials by phlebioid fungal species. Bioresour Technol 225:254–261. https://doi.org/10.1016/j.biortech.2016.11.082
Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428
Nagy LG, Riley R, Bergmann PJ, Krizsán K, Martin FM, Grigoriev IV, Cullen D, Hibbett DS (2017) Genetic bases of fungal white rot wood decay predicted by phylogenomic analysis of correlated gene-phenotype evolution. Mol Biol Evol 34:35–44. https://doi.org/10.1093/molbev/msw238
Nakasone KK, Sytsma KJ (1993) Biosystematic studies on Phlebia acerina, P. rufa, and P. radiata in North America. Mycologia 85:996–1016. https://doi.org/10.2307/3760683
Niranjane AP, Madhou P, Stevenson TW (2007) The effect of carbohydrate carbon sources on the production of cellulase by Phlebia gigantea. Enzym Microb Technol 40:1464–1468. https://doi.org/10.1016/j.enzmictec.2006.10.041
Okamoto K, Imashiro K, Akizawa Y, Onimura A, Yoneda M, Nitta Y, Maekawa N, Yanase H (2010) Production of ethanol by the white-rot basidiomycetes Peniophora cinerea and Trametes suaveolens. Biotechnol Lett 32:909–913. https://doi.org/10.1007/s10529-010-0243-7
Patyshakuliyeva A, Mäkelä MR, Sietiö O-M, de Vries RP, Hildén KS (2014) An improved and reproducible protocol for the extraction of high quality fungal RNA from plant biomass substrates. Fungal Genet Biol 72:201–206. https://doi.org/10.1016/j.fgb.2014.06.001
Ruiz-Dueñas FJ, Lundell T, Floudas D, Nagy LG, Barrasa JM, Hibbett DS, Martínez AT (2013) Lignin-degrading peroxidases in Polyporales: an evolutionary survey based on 10 sequenced genomes. Mycologia 105:1428–1444. https://doi.org/10.3852/13-059
Rytioja J, Hildén K, Hatakka A, Mäkelä MR (2014a) Transcriptional analysis of selected cellulose-acting enzymes encoding genes of the white-rot fungus Dichomitus squalens on spruce wood and microcrystalline cellulose. Fungal Genet Biol 72:91–98. https://doi.org/10.1016/j.fgb.2013.12.008
Rytioja J, Hildén K, Yuzon J, Hatakka A, de Vries RP, Mäkelä MR (2014b) Plant-polysaccharide-degrading enzymes from basidiomycetes. Microbiol Mol Biol Rev 78:614–649. https://doi.org/10.1128/MMBR.00035-14
Salavirta H, Oksanen I, Kuuskeri J, Mäkelä M, Laine P, Paulin L, Lundell T (2014) Mitochondrial genome of Phlebia radiata is the second largest (156 kbp) among fungi and features signs of genome flexibility and recent recombination events. PLoS One 9:e97141. https://doi.org/10.1371/journal.pone.0097141
Saloheimo M, Niku-Paavola M-L, Knowles JKC (1991) Isolation and structural analysis of the laccase gene from the lignin-degrading fungus Phlebia radiata. J Gen Microbiol 137:1537–1544. https://doi.org/10.1099/00221287-137-7-1537
Shary S, Kapich AN, Panisko EA, Magnuson JK, Cullen D, Hammel KE (2008) Differential expression in Phanerochaete chrysosporium of membrane-associated proteins relevant to lignin degradation. Appl Environ Microbiol 74:7252–7257. https://doi.org/10.1128/AEM.01997-08
Skyba O, Cullen D, Douglas CJ, Mansfield SD (2016) Gene expression patterns of wood decay fungi Postia placenta and Phanerochaete chrysosporium are influenced by wood substrate composition during degradation. Appl Environ Microbiol 82:4387–4400. https://doi.org/10.1128/AEM.00134-16
Steiger MG, Mach RL, Mach-Aigner AR (2010) An accurate normalization strategy for RT-qPCR in Hypocrea jecorina (Trichoderma reesei). J Biotechnol 145:30–37. https://doi.org/10.1016/j.jbiotec.2009.10.012
Stewart P, Cullen D (1999) Organization and differential regulation of a cluster of lignin peroxidase genes of Phanerochaete chrysosporium. J Bacteriol 181:3427–3432
Vaaje-Kolstad G, Westereng B, Horn SJ, Liu Z, Zhai H, Sorlie M, Eijsink VGH (2010) An oxidative enzyme boosting the enzymatic conversion of recalcitrant polysaccharides. Science 330:219–222. https://doi.org/10.1126/science.1192231
Vallim MA, Janse BJ, Gaskell J, Pizzirani-Kleiner AA, Cullen D (1998) Phanerochaete chrysosporium cellobiohydrolase and cellobiose dehydrogenase transcripts in wood. Appl Environ Microbiol 64:1924–1928
van den Brink J, de Vries RP (2011) Fungal enzyme sets for plant polysaccharide degradation. Appl Microbiol Biotechnol 91:1477–1492. https://doi.org/10.1007/s00253-011-3473-2
Vanden Wymelenberg A, Gaskell J, Mozuch M, Kersten P, Sabat G, Martinez D, Cullen D (2009) Transcriptome and secretome analyses of Phanerochaete chrysosporium reveal complex patterns of gene expression. Appl Environ Microbiol 75:4058–4068. https://doi.org/10.1128/AEM.00314-09
Vogel C, Marcotte EM (2012) Insights into the regulation of protein abundance from proteomic and transcriptomic analyses. Nat Rev Genet 13:227–232. https://doi.org/10.1038/nrg3185
Vu VV, Beeson WT, Span EA, Farquhar ER, Marletta MA (2014) A family of starch-active polysaccharide monooxygenases. Proc Natl Acad Sci U S A 111:13822–13827. https://doi.org/10.1073/pnas.1408090111
Yoshida M, Igarashi K, Kawai R, Aida K, Samejima M (2004) Differential transcription of β-glucosidase and cellobiose dehydrogenase genes in cellulose degradation by the basidiomycete Phanerochaete chrysosporium. FEMS Microbiol Lett 235:177–182. https://doi.org/10.1111/j.1574-6968.2004.tb09584.x
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The authors thank Jaana Kuuskeri for the aid with gene annotations and primer design.
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This study was funded by the Academy of Finland project grant (no. 285676, to TL).
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MM, HM, and TL designed the study. NR and MM carried out the experiments. MM, NR, and HM analyzed the data. MM, HM, and TL interpreted the data. MM and TL wrote the manuscript. All authors read and approved the final manuscript.
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Mäkinen, M.A., Risulainen, N., Mattila, H. et al. Transcription of lignocellulose-decomposition associated genes, enzyme activities and production of ethanol upon bioconversion of waste substrate by Phlebia radiata. Appl Microbiol Biotechnol 102, 5657–5672 (2018). https://doi.org/10.1007/s00253-018-9045-y
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DOI: https://doi.org/10.1007/s00253-018-9045-y