ISOLATION OF PURE CULTURES OF WOOD-DESTRUCTING FUNGI AND THEIR MOLECULAR CHARACTERIZATION FOR BIOTECHNOLOGICAL APPLICATION
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Authors
A.A. Saparbekova
M. Auezov South Kazakhstan State University, Shymkent, Kazakhstan
A.B. Altekey
M. Auezov South Kazakhstan State University, Shymkent, Kazakhstan
B.R. Silvia
University of Valladolid, Valladolid, Spain
A.D. Matchanov
Academician Sadykov Institute of Bioorganic Chemistry, Tashkent, Uzbekistan
G. Kantureyeva
M. Auezov South Kazakhstan State University, Shymkent, Kazakhstan
G. Kaldybekova
M. Auezov South Kazakhstan State University, Shymkent, Kazakhstan
Abstract
This paper presents a method for isolating pure cultures of wood-destroying fungi from the surface of decomposing wood collected in the Asanbay Askarov Arboretum (Shymkent, Kazakhstan), performing their microscopic examination and molecular identification using polymerase chain reaction (PCR) and ITS region sequencing. The aim of the study is to investigate the species diversity of wood-destroying fungi and assess their potential for biotechnological applications. Five species were identified: Aspergillus spelaeus, Aspergillus niger, Penicillium sizovae, Cladosporium cladosporioides, and Alternaria sp. Isolation was performed using enrichment cultures, microscopy with methylene blue staining, and identification by PCR and sequencing. The use of PCR and sequencing enabled accurate species identification and proposes new approaches for monitoring and studying ecosystem processes associated with wood surfaces. Ligninolytic activity was confirmed using the Azure-B assay. The data obtained highlight the high diversity of fungi and their potential for wood waste processing and bioremediation. The results showed a high diversity of wood-destroying fungi on wood surfaces, confirming the complexity of the microbiome in these ecosystems.
Keywords
wood-destroying fungi, wood, identification, lignin, political activism, bioremediation
Article Details
References
Kirk TK, Farrell RL. Enzymatic "combustion": the microbial degradation of lignin. Annu Rev Microbiol. 1987;41:465–505. doi:10.1146/annurev.mi.41.100187.002341
Leonowicz A, Matuszewska A, Luterek J, et al. Biodegradation of lignin by white rot fungi. Fungal Genet Biol. 1999;27(2–3):175–85. doi:10.1006/fgbi.1999.1150
Cui T, Yuan B, Guo H, et al. Enhanced lignin biodegradation by consortium of white rot fungi: microbial synergistic effects and product mapping. Biotechnol Biofuels. 2021;14:162. doi:10.1186/s13068-021-02011-y
Kamimura N, Takahashi K, Mori K, et al. Advances in microbial lignin degradation and its applications. Curr Opin Biotechnol. 2019;56:179–86. doi:10.1016/j.copbio.2018.11.011
Saparbekova AA, Altekey AB, et al. Biotechnological methods in pulp and paper production from an alternative source of raw materials - the savior of the ecosystem. Appl Environ Biotechnol. 2023;8(2):1–9. doi:10.26789/AEB.2023.02.001
Janusz G, Pawlik A, Sulej J, et al. Lignin degradation: microorganisms, enzymes involved, genomes analysis and evolution. FEMS Microbiol Rev. 2017;41(6):941–62. doi:10.1093/femsre/fux049
Altekey AA, Saparbekova AA, et al. Biodegradation of straw lignin by culture of the fungus Schizophyllum commune isolated from the tree surface. J Microbiol Virol. 2023;1(40):127–38. doi:10.53729/MV-AS.2023.01.08
Harms H, Schlosser D, Wick LY. Untapped potential: exploiting fungi in bioremediation of hazardous chemicals. Nat Rev Microbiol. 2011;9(3):177–92. doi:10.1038/nrmicro2519
R. Rapley The Nucleic Acids Protocols Handbook. Human Press. Totowa, New Jersey. – 2000.- 1002p.
Badotti F, de Oliveira FS, Garcia CF, et al. Effectiveness of ITS and sub-regions as DNA barcode markers for the identification of Basidiomycota (Fungi). BMC Microbiol. 2017;17:42. doi:10.1186/s12866-017-0958-x
Clarridge JE 3rd. Impact of 16S rRNA gene sequence analysis for identification of bacteria on clinical microbiology and infectious diseases. Clin Microbiol Rev. 2004;17(4):840-62.
Altekey A. Induction of ligninolytic enzymes of the fungus Schizophyllum commune. Eurasian J Ecol. 2024;78(1):45-53. ISSN 2617-7358. doi:10.26577/EJE.2024.v78.i1.08
Buzina W, et al. Development of molecular methods for identification of Schizophyllum commune from clinical samples. J Clin Microbiol. 2001;39(7):2391-6. doi:10.1128/JCM.39.7.2391-2396.2001
Kumar VP, Sridhar M, Rao RG. Biological depolymerization of lignin using laccase harvested from the autochthonous fungus Schizophyllum commune employing various production methods and its efficacy in augmenting in vitro digestibility in ruminants. Sci Rep. 2022;12(1):11170. doi:10.1038/s41598-022-15211-9
Toju H, et al. High-coverage ITS primers for the DNA-based identification of Ascomycetes and Basidiomycetes in environmental samples. Lespinet O, editor. PLoS One. 2012;7(7):e40863. doi:10.1371/journal.pone.0040863
Costa S, et al. Lignin biodegradation in pulp-and-paper mill wastewater by selected white rot fungi. Water. 2017;9(12):935. doi:10.3390/w9120935
Mytishchi branch, et al. Biodegradation of wood by wood-destroying fungi enzyme complexes. Forestry Bull. 2019;135:95-100. doi:10.18698/2542-1468-2019-5-95-100
Bhardwaj N, Kumar B, Agrawal K, et al. Current perspective on production of fungal cellulases under solid-state fermentation using agro-industrial by-products and their applications. 3 Biotech. 2020;10(6):227. doi:10.1007/s13205-020-02240-
Costa S, Dedola DG, Pellizzoni M, et al. Lignin biodegradation in pulp-and-paper mill wastewater by selected white rot fungi. Water. 2017;9(12):935. doi:10.3390/w9120935
Rouches E, Herpoël-Gimbert I, Steyer JP, et al. Potential of white-rot fungi for sustainable bioethanol production from lignocellulosic materials. Renew Sustain Energy Rev. 2016;64:1272–81. doi:10.1016/j.rser.2016.06.053
Zhang J, Liu X, Liu Y, et al. Degradation of lignin by Cladosporium cladosporioides: insights into enzymatic mechanisms. Appl Microbiol Biotechnol. 2018;102(15):6543–54. doi:10.1007/s00253-018-9105-6
Sigoillot JC, Berrin JG, Bey M, et al. Fungal strategies for lignin degradation. Fungal Biol Rev. 2012;26(4):203–13. doi:10.1016/j.fbr.2012.05.002