Cellulose hydrolysis by Trichoderma viride enzyme complex in the presence of sodium fluoride: effect of substrate structure and cellulase sorption activity
https://doi.org/10.21285/2227-2925-2020-10-2-261-273
Abstract
The dependence of cellulase activity of the Trichoderma viride fungus on three factors – substrate type, enzyme sorption properties and sodium fluoride (NaF) concentration – was studied. The four studied water-insoluble substrates were based on two native (birch wood and wheat straw) and two model (microcrystalline cellulose and filter paper) materials. In addition, a water-soluble Na-carboxymethyl-cellulose substrate specific for endoglucanase activity was studied. The T. viride cellulase complex from the “Celloviridin G3x” preparation was separated into fractions differing in sorption properties using the affinity chromatography method. The cellulase activity of the initial complex and obtained fractions was measured with respect to all substrates at different concentrations of NaF ranging from 0 to 1000 mg/L. Qualitative differences in the protein composition of the preparation and fractions were studied by SDS electrophoresis. With the introduction of NaF (100–500 mg/L), the initial cellulase complex activity increased with respect to all substrates. The hydrolysis rate of model samples was observed to increase by up to 100 %. Comparison of chromatographic fraction activity relative to model substrates demonstrated the NaF to have the opposite effect on the catalytic and sorption properties of cellulases. The activity of high affinity fractions containing cellobiohydrolases and low molecular weight endoglucanases in the presence of fluoride decreased by 35–90 % in relation to all substrates. Although weakly adsorbing fractions (endoglucanases I, II + cellobiasis) under the same conditions were inhibited by 15–20 % with respect to cellulose with a high degree of crystallinity (MCC), these were activated 1.3–3 times slower with respect to a water-soluble substrate (Na-CMC) and a substrate with less crystallinity (filter paper). Thus, the total effect of fluorides on the enzymatic hydrolysis of the plant substrate is determined by the crystallinity degree of the cellulose substrate, as well as by the ratio of stronglyand weakly-sorbed components in the cellulase complex. Possible molecular mechanisms of these effects are additionally proposed.
About the Authors
E. R. Chashina
Irkutsk State University
Russian Federation
Master Student
1, Karl Marx St., Irkutsk, 664003, Russian Federation
Russian Federation
Master Student
1, Karl Marx St., Irkutsk, 664003, Russian Federation
Z. A. Efremenko
Irkutsk State University
Russian Federation
Postgraduate Student
1, Karl Marx St., Irkutsk, 664003, Russian Federation
Russian Federation
Postgraduate Student
1, Karl Marx St., Irkutsk, 664003, Russian Federation
V. P. Salovarova
Irkutsk State University
Russian Federation
Dr. Sci. (Biology), Professor, Head of the Department of Physico-Chemical Biology, Bioengineering and Bioinformatics
1, Karl Marx St., Irkutsk, 664003, Russian Federation
Russian Federation
Dr. Sci. (Biology), Professor, Head of the Department of Physico-Chemical Biology, Bioengineering and Bioinformatics
1, Karl Marx St., Irkutsk, 664003, Russian Federation
D. E. Gavrikov
Irkutsk State University
Russian Federation
Cand. Sci. (Biology), Associate Professor, Department of Natural Sciences, Pedagogical Institute
1, Karl Marx St., Irkutsk, 664003, Russian Federation
Russian Federation
Cand. Sci. (Biology), Associate Professor, Department of Natural Sciences, Pedagogical Institute
1, Karl Marx St., Irkutsk, 664003, Russian Federation
A. A. Pristavka
Irkutsk State University
Russian Federation
Cand. Sci. (Biology), Associate Professor, Department of Physico-Chemical Biology, Bioengineering and Bioinformatics
1, Karl Marx St., Irkutsk, 664003, Russian Federation
Russian Federation
Cand. Sci. (Biology), Associate Professor, Department of Physico-Chemical Biology, Bioengineering and Bioinformatics
1, Karl Marx St., Irkutsk, 664003, Russian Federation
References
1. Kanitskaya LV, Kolmogorov AV. Impact of gas emissions from aluminum production on the environment. Uspekhi sovremennogo estestvoznaniya. 2009;8:17–18. (In Russian)
2. Evdokimova GA, Mozgova NP. Soil and plant pollution assessment in the zone of exposure to gas-air emissions from an aluminum smelter. Teoreticheskaya i prikladnaya ekologiya = Theoretical and Applied Ecolody. 2015;4:64-68. (In Russian) https://doi.org/10.25750/1995-4301-2015-4-064-068
3. Berseneva OA, Salovarova VР, Pristavka AA, Milentyev VA. Specific structure of the soil mycocenoses in grey forest soil of pribaikalye subject to influence of emissions of Irkutsk aluminium plant. Vestnik RUDN. Seriya Ekologiya i bezopasnost' zhiznedeyatel'nosti = RUDN Journal of Ecology and Life Safety. 2010;1:24–29. (In Russian)
4. Evdokimova GA, Korneykova MV, Lebedeva EV. Micromycete communities in soils in an aluminum smelter impact area. Mikologiya i Fitopatologiya = Mycology and Phytopathology. 2007;41(1):20–28. (In Russian)
5. Blagodatskaya EV, Semenov MV, Yakushev AV. Activity and biomass of soil microorganisms in changing environmental conditions. Moscow: Tovarishchestvo nauchnykh izdanii KMK; 2016. 243 p. (In Russian)
6. Katiyar P, Pandey N, Sahu KK. Biological approaches of fluoride remediation: potential for environmental clean-up. Environmental Science and Pollution Research. 2020;27:13044–13055. https://doi.org/10.1007/s11356-020-08224-2
7. Evdokimova GA, Zenkova IV, Mozgova NP. Soil and soil microbiota under fluorine pollution. Apatity: Kol'skii nauchnyi tsentr RAN; 2005. 135 p. (In Russian)
8. Makarova AP, Bukovskaja NE, Naprasnikova EV. The Impact of Aerotechnogenic Emissions from Aluminum Smelters in the Irkutsk Region on the Soil Microbiota. Izvestiya Irkutskogo gosudarstvennogo universiteta. Seriya Biologiya. Ecologiya = The Bulletin of Irkutsk State University. Series Biology. Ecology. 2017;19:57–62. (In Russian)
9. Gornostaeva EA, Fuks SL. The effect of fluorinated compounds on living organisms (review). Teoreticheskaya i prikladnaya ekologiya = Theoretical and Applied Ecolody. 2017;1:14–24. (In Russian)
10. Pristavka AA, Popova IV. Influence of sodium fluoride on enzymatic activity of fungal cellulases. Izvestiya Vuzov. Prikladnaya Khimiya i Biotekhnologiya = Proceedings of Universities. Applied Chemistry and Biotechnology. 2015;1:36–46. (In Russian)
11. Klyosov AA, Rabinowitch ML, Sinitsyn AP, Churilova IV, Grigorash SYu. Enzymatic hydrolysis of cellulose. I. Activity and composition of cellulase complexes from various sources. Bioorganicheskaya Khimiya = Russian Journal of Bioorganic Chemistry. 1980;6(8):1225–1242. (In Russian)
12. Sinitsyn AP, Osipov DO, Rozhkova AM, Bushina EV, Dotsenko GS, Sinitsyna OA, et al. The production of highly effective enzyme complexes of cellulases and hemicellulases based on the Penicillium verruculosum strain for the hydrolysis of plant raw materials. Applied Biochemistry and Microbiology. 2014;50(8):761–772. https://doi.org/10.1134/S0003683814080055
13. Beheraa BC, Sethib BK, Mishra RR, Dutta SK, Thatoi HN. Microbial cellulases – Diversity & biotechnology with reference to mangrove environment: A review. Journal of Genetic Engineering and Biotechnology. 2017;15(1):197–210. https://doi.org/10.1016/j.jgeb.2016.12.001
14. Thygesen A, Oddershede J, Lilholt H, Thomsen AB, Stahl K. On the determination of crystallinity and cellulose content in plant fibres. Cellulose. 2005;12(6):563–576. https://doi.org/10.1007/s10570-005-9001-8
15. Kaschuk JJ, Frollini E. Effects of average molar weight, crystallinity, and hemicelluloses content on the enzymatic hydrolysis of sisal pulp, filter paper, and microcrystalline cellulose. Industrial Crops and Products. 2018;15:280–289. https://doi.org/10.1016/j.indcrop.2018.02.011
16. Ahvenainen P, Kontro I, Svedström K. Comparison of sample crystallinity determination methods by X-ray diffraction for challenging cellulose I materials. Cellulose. 2016;23(2):1073–1086. https://doi.org/10.1007/s10570-016-0881-6
17. Ioelovich MYa. Models of supramolecular structure and properties of cellulose. Vysokomolekulyarnye soedineniya, Seriya A. = Polymer Science, Series A. 2016;58:925–943. (In Russian) https://doi.org/10.1134/S0965545X16060109
18. Ghose TK. Measurement of cellulase activity. Pure and Applied Chemistry. 1987;59:257–268. https://doi.org/10.1351/pac198759020257
19. Rabinowitch ML, Klyosov AA, Berezin IV. Kinetics of the action of cellulolytic enzymes from Geotrihum candidum. Viscometric analysis of the kinetics of carboxymethyl cellulose hydrolysis. Bioorganicheskaya khimiya = Russian Journal of Bioorganic Chemistry 1977;З(3):405–414. (In Russian)
20. Klyosov AA, Mitkevich OV, Sinitsyn AP. Role of the activity and adsorption of cellulases in the efficiency of the enzymic hydrolysis of amorphous and crystalline cellulose. Biochemistry. 1986:25(3):540–542 https://doi.org/10.1021/bi00351a003
21. Saloheimo M, Nakari-Setälä T, Tenkanen M, Penttilä M. cDNA cloning of a Trichoderma reesei cellulase and demonstration of endoglucanase activity by expression in yeast. European Journal of Biochemistry. 1997;249(2):584–591. https://doi.org/10.1111/j.1432-1033.1997.00584.x
22. Pristavka AA, Salovarova VP, Zacchi G, Berezint IV, Rabinovich ML. Enzyme Recovery in High-Solids Enzymatic Hydrolysis of SteamPretreated Willow: Requirements for the Enzyme Composition. Applied Biochemistry and Microbiology. 2000;36(3):237–244. https://doi.org/10.1007/BF02742572
23. Aghajari N, Feller G, Gerday C, Haser R. Structural basis of α-amylase activation by chloride. Protein Science. 2002;11(6):1435–1441. https://doi.org/10.1110/ps.0202602 ·
24. Linder M, Mattinen ML, Kontteli M, Lindeberg G, Stihlberg J, Drakenberg T., et al. Identification of functionally important amino acids in the cellulose-binding domain of Trichoderma reesei cellobiohydrolase I. Protein Science. 1995;4(6):1056–1064. https://doi.org/10.1002/pro.5560040604
25. Evdokimova GA, Mozgova NP, Pereverzev VN. Transformation of plant residues in the soil of a zone exposed to emissions from an aluminum smelter. Eurasian Soil Science. 2013;46(8):908–917. https://doi.org/10.1134/S1064229313060033
Review
For citations:
Chashina E.R., Efremenko Z.A., Salovarova V.P., Gavrikov D.E., Pristavka A.A. Cellulose hydrolysis by Trichoderma viride enzyme complex in the presence of sodium fluoride: effect of substrate structure and cellulase sorption activity. Proceedings of Universities. Applied Chemistry and Biotechnology. 2020;10(2):261-273. (In Russ.) https://doi.org/10.21285/2227-2925-2020-10-2-261-273
Views:
693
Email this article 