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Superior cellulolytic activity of Trichoderma guizhouense on raw wheat straw

  • Marica Grujić
  • Biljana DojnovEmail author
  • Ivana Potočnik
  • Lea Atanasova
  • Bojan Duduk
  • Ewald Srebotnik
  • Irina S. Druzhinina
  • Christian P. Kubicek
  • Zoran Vujčić
Original Paper
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Abstract

Lignocellulosic plant biomass is the world’s most abundant carbon source and has consequently attracted attention as a renewable resource for production of biofuels and commodity chemicals that could replace fossil resources. Due to its recalcitrant nature, it must be pretreated by chemical, physical or biological means prior to hydrolysis, introducing additional costs. In this paper, we tested the hypothesis that fungi which thrive on lignocellulosic material (straw, bark or soil) would be efficient in degrading untreated lignocellulose. Wheat straw was used as a model. We developed a fast and simple screening method for cellulase producers and tested one hundred Trichoderma strains isolated from wheat straw. The most potent strain—UB483FTG2/ TUCIM 4455, was isolated from substrate used for mushroom cultivation and was identified as T. guizhouense. After optimization of growth medium, high cellulase activity was already achieved after 72 h of fermentation on raw wheat straw, while the model cellulase overproducing strain T. reesei QM 9414 took 170 h and reached only 45% of the cellulase activity secreted by T. guizhouense. Maximum production levels were 1.1 U/mL (measured with CMC as cellulase substrate) and 0.7 U/mL (β-glucosidase assay). The T. guizhouense cellulase cocktail hydrolyzed raw wheat straw within 35 h. Our study shows that screening for fungi that successfully compete for special substrates in nature will lead to the isolation of strains with qualitatively and quantitatively superior enzymes needed for their digestion which could be used for industrial purposes.

Keywords

Cellulases Cellulase enzyme complex Wheat straw Trichoderma Trichoderma guizhouense Trichoderma reesei 
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Notes

Acknowledgements

This study was supported by a grant from the Serbian Ministry of Education, Science and Technological Development (Project Grant Number OI172048). The work in TU Wien was supported by the Austrian Science Fund (FWF) Project Number P 25613 B20 to ISD. We thank Dr. Komal Chenthamara for help with in situ data analysis. We thank Jasna Bojčevski for English correction.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11274_2019_2774_MOESM1_ESM.tif (1.5 mb)
Fig. S1. Screening for efficient cellulase producers among Trichoderma isolates. Total liquefaction of the medium (level 4 from Table 1) occurred within 25 h for isolate T. guizhouense TG2 and first minor changes in viscosity (level 2 from Table 1) occurred within 25 h for T. reesei QM9414. (TIF 1509 kb)
11274_2019_2774_MOESM2_ESM.tif (11.7 mb)
Fig. S2. Position of Trichoderma strain TG2 in the tef1 Bayesian phylogenetic tree. Branching points supported by posterior probabilities higher than 0.94 are indicated by grey dots. Arrowindicates branching that leads to the Trichoderma harzianum species clade. Subclade names within the Trichoderma harzianum species complex, including strain TG2 are shown in bold. Tailing digits correspond to GenBank accession numbers. (TIF 12006 kb)
11274_2019_2774_MOESM3_ESM.tif (6.2 mb)
Fig. S3. Optimization of cellulase complex production by T. guizhouense TG2. Influence of a) substrate amount; b) agitation (CC- combined conditions); c) inoculation (SS-spore suspension; MD-mycelial disk; I1-inoculum1; I2-inoculum 2; I3-inoculum 3); and d) nitrogen sources, on production levels. Enzyme activities are presented as U/mL, and are displayed as the mean of three independent assays ± SEM (all below 5%). (TIF 6329 kb)
11274_2019_2774_MOESM4_ESM.pdf (60 kb)
Table S1. Optimization of the production of cellulase enzyme complex by TG2 with untreated wheat straw as substrate. (PDF 60 kb)
11274_2019_2774_MOESM5_ESM.pdf (117 kb)
Table S2. Comparison of maximal cellulase production (U/mL) of T. guizhouense TG2 and T. reesei QM9414 on raw wheat straw substrate. Enzyme activities are presented as U/mL, and are displayed as the mean of three independent assays ± SEM (all below 5%). (PDF 116 kb)

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Marica Grujić
    • 1
  • Biljana Dojnov
    • 2
    Email author
  • Ivana Potočnik
    • 3
  • Lea Atanasova
    • 4
    • 7
  • Bojan Duduk
    • 3
  • Ewald Srebotnik
    • 5
  • Irina S. Druzhinina
    • 4
    • 6
  • Christian P. Kubicek
    • 4
    • 8
  • Zoran Vujčić
    • 1
  1. 1.Department of Biochemistry, Faculty of ChemistryUniversity of BelgradeBelgradeSerbia
  2. 2.Department of Chemistry, Institute of Chemistry, Technology and MetallurgyUniversity of BelgradeBelgradeSerbia
  3. 3.Laboratory of Applied PhytopathologyInstitute of Pesticides and Environmental ProtectionBelgradeSerbia
  4. 4.Microbiology and Applied Genomics Group, Research Division of Biochemical TechnologyInstitute of Chemical, Environmental and Bioscience Engineering (ICEBE), TU WienViennaAustria
  5. 5.Bioresource Technology Group, Research Division of Bioresources and Plant ScienceInstitute of Chemical, Environmental and Bioscience Engineering (ICEBE), TU WienViennaAustria
  6. 6.Jiangsu Provincial Key Lab of Organic Solid Waste UtilizationNanjing Agricultural UniversityNanjingChina
  7. 7.Institute of Food Technology, University of Natural Resources and Life Sciences (BOKU)ViennaAustria
  8. 8.ViennaAustria

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