RESEARCH PAPER
Application of semi-quantitative and quantitative methods for the selection of cellulolytic filamentous fungi isolated from pulp mill materials
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Department of Biotechnology, Kazimierz Wielki University, Bydgoszcz, Poland
Submission date: 2016-04-11
Final revision date: 2016-06-03
Acceptance date: 2016-06-09
Publication date: 2016-11-02
BioTechnologia 2016;97(3):169-178
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ABSTRACT
A semi-quantitative method (the enzymatic activity index, EI) and a quantitative method (the amount of reducing sugars, RS) have been used for selecting cellulolytic microorganisms. The analysis was performed with 90 isolates of filamentous fungi selected from pulp mill materials. In both methods 1% low viscosity carboxymethylcellulose (1% CMC l.v.) was used as a substrate. For n = 90, no correlation between the parameters has been found.
A further analysis was carried out for 12 selected isolates: 6 with the highest RS values and 6 with the highest EI values. For the chosen fungi, also cultivated on 1% CMC l.v. medium, the following parameters have been determined: the total cellulase activity (filter paper assay, FPase), endoglucanase (CMCase) activity, β –glucosidase (CBase) activity, and the changes in the culture medium viscosity (DV). The statistical analysis revealed a negative, moderately marked correlation between EI and RS and clearly marked positive correlations for two pairs: RS-FPase and DV-FPase. A high positive correlation between RS and DV has also been observed. Correlations between EI, RS, DV, CMCase, and CBase activities were weak. It was shown that the quantitative parameter RS is more suitable than EI for the evaluation of the total cellulolytic activity of filamentous fungi. The application of CMC as the only source of carbon in the culture media and the subsequent determination of EI or RS has failed to provide the means for selecting fungi with a high endoglucanase (CMCase) activity.
REFERENCES (28)
1.
Ahmed S., Bashir A., Saleem H., Saadia M., Jamil A. (2009) Production and purification of cellulose degrading enzymes from a filamentous fungus Trichoderma harzianum. Pak. J. Bot. 41 (3): 1411-1419.
2.
Alvira P., Tomás-Pejó E., Ballesteros M., Negro M.J. (2010) Pretreatment technologies for an efficient bioethanol production process based on enzymatic hydrolysis – an overview. Bioresource Technol. 101: 4851-4861.
3.
Bhat M.K. (2000) Cellulases and related enzymes in biotechnology. Biotechnol. Adv. 18: 355-383.
4.
Bradford M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal. Biochem. 72: 248-254.
5.
Cherry J.R., Fidantsef A.L. (2003) Directed evolution of industrial enzymes: an update. Curr. Opin. Biotechnol. 14: 438-443.
6.
Crous P.W., Verkley G.J.M., Groenewald J.Z., Samson R.A. (2009) Fungal biodiversity. CBS-KNAW Fungal Biodiversity Centre, Utrecht, Netherlands.
7.
Damaso M.C.T., da Costa Terzi S., Farias A.X., Pereira de Oliveira A.C., Fraga M.E., Couri S. (2012) Selection of cellulolytic fungi isolated from diverse substrates. Braz. Arch. Biol. Techn. 55 (4): 513-520.
8.
Fassatiova O. (1983) Filamentous fungi in technical microbiology (in polish). Wyd. Naukowo-Techniczne, Warsaw, Poland.
9.
Florencio C., Couri S., Farinas C.S. (2012) Correlation between agar plate screening and solid-state fermentation for the prediction of cellulase production by Trichoderma strains. Enzyme Res., 1-7.
10.
François J.M., Guais O. (2011) Genomic and proteomic analyses provide insights into the potential of filamentous fungi for biomass degradation. In: Mycofactories. Ed. Leitão A.L., Portugal: Bentham Science Publishers: 45-56.
11.
Ghose T.K. (1987) Measurement of cellulose activities. Pure & App. Chem. 59(2): 257-268.
12.
Goldbeck R., Andrade C.C.P., Pereira G.A.G., Filho M. (2012) Screening and identification of cellulase producing yeastlike microorganisms from Brazilian biomes. Afr. J. Biotechnol. 11(53): 11595-11603.
13.
Huggett A.S.C., Nixon D.A. (1955) Glucose oxidase method for measurement of glucose. Biochem. J. 66: 12-19.
14.
Ishikawa Y., Saka S. (2001) Chemical conversion of cellulose as treated in supercritical methanol. Cellulose 8: 189-195.
15.
Jahangeer S., Khan N., Jahangeer S., Sohail M., Shahzad S., Ahmed A. (2005) Screening and characterization of fungal cellulases isolated from the native environmental source. Pak. J. Bot. 37(3): 739-748.
16.
Kuhad R.C., Gupta R., Singh A. (2011) Microbial cellulases and their industrial applications. Enzyme Res. 1-10.
17.
Kwaśna H., Chełkowski J., Zajkowski P. (1991) Fungi, vol. 22 – in polish. PAN, Warszawa-Kraków, Poland.
18.
Lopes F., Motta F., Andrade C.C.P., Rodrigues M.I., Maugeri- Filho F. (2011) Thermo-stable xylanases from non conventional yeasts. J. Microbial. Biochem. Technol. 3(3): 36-42.
19.
Miller G.L. (1956) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem. 1: 426-428.
20.
Pérez J., Munoz-Moraleda A. (2011) Fungal ligninocellulolytic enzymes: applications in biodegradation and bioconversion. In: Mycofactories. Ed Leitão A.L., Portugal: Bentham Sci. Publ.: 28-44.
21.
Ruegger M.J.S., Tauk-Tornisielo S.M. (2004) Cellulase activity of fungi isolated from soil of the Ecological Station of Juréia-Itatins, São Paulo, Brazil. Brazil. J. Bot. 27: 205- 211.
22.
Russel S., Górska E.B., Wyczółkowski A.I. (2005) Enzymes taking part in hydrolysis of cellulose. Acta Agrophysica 3: 27-36.
23.
Samson R.A., Frisvad J.C. (2004) Penicillium subgenus Penicillium: new taxonomic schemes and mycotoxins and other extrolites. Stud. Mycol. 449: 1-174.
24.
Sazci A., Erenler K., Radford A. (1986) Detection of cellulolytic fungi by using Congo red as an indicator: a comparative study with the dinitrosalicylic acid reagent method. J. Appl. Bacteriol. 61(6): 559-562.
25.
Shuangqi T., Zhenyu W., Ziluan F., Lili Z., Jichang W. (2011) Determination methods of cellulase activity. Afr. J. Biotechnol. 10(37): 7122-7125.
26.
Sun Y., Cheng J. (2002) Hydrolysis of lignocellulosic materials for ethanol production-an overview. Bioresour. Technol. 83: 1-11.
27.
Teather R., Wood P. (1982) Use of Congo Red-polysaccharide interactions in enumeration and characterization of cellulolytic bacteria from the bovine rumen. Appl. Environ. Microb. 43(4): 777-780.
28.
Zhang Y.-H.P., Himmel M.E., Mielenz J.R. (2006) Outlook for cellulase improvement: screening and selection strategies. Biotechnol. Adv. 24: 452-481.