Enzyme activity of cellulases produced by Moniliophtora perniciosa, the causing factor of “witch–broom” in cacao plants

Authors

  • Carlos Eduardo Sampaio Guedes Universidade Federal da Bahia
  • Rita Cajazeira Alvarez Universidade Federal da Bahia
  • Eduardo Aquino Ximenes Laboratory of Renewable Resources Engineering, Purdue University, Agricultural Engineering Department
  • Luís César Paulillo Universidade Federal da Bahia
  • Suzana Telles Cunha Lima Universidade Federal da Bahia
  • Luciana Veiga Barbosa

Abstract

Abstract : Cellulases are applied in food and textile industry, in the production of alcoholic beverages, bioconversion of agronomic residues and, increasingly, for production of biofuels. The fungi of cacao plants are very resistant and difficult to combat. On the other hand, enzymes produced by them have numerous biotechnological applications. The present work aimed to evaluate isolates of Moniliophtora perniciosa representatives from the Reconcavo region of Bahia, in Brazil,  regarding the production of cellulases. Thirty two  M. perniciosa strains were evaluated while growing in the presence of cellulose as the only carbon source. They were initially screened using the Congo red assay, following halo formation, after growth in solid medium containing one of the substrates mentioned before. Of the tested strains, 948F showed the greatest potential for the hydrolysis of non-crystalline cellulose. This strain was further tested for its ability to produce cellulases in minimal liquid medium in the presence of carboximetil cellulose (CMC) as the only carbon source. The maximal growth range was between 168 and 192 h. The highest activity values were observed between 240 and 246 h, maximum value being 10.5 U/mL x    10-3. The zymogram analysis showed the presence of three bands with endoglucanase activity. Although the results obtained are preliminary, they may open the way for future investigations of this strain aiming optimization of conditions for production of cellulases and other hydrolytic enzymes with potential biotechnological applications.

 Key words: Theobroma cacao, Endoglucanases, Hydrolytic enzymes.

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References

ALFENAS, A. C. Eletroforese de izoenzimas e proteínas afins: fundamentos aplicações em plantas e microrganismos. Viçosa: UFV, 1998.

AZEVEDO, M.O.; POÇAS-FONSECA, M.J. Melhoramento genético de fungos celulolíticos. In: DE MELO, I.S.; VALADARES-INGLIS, M.C.; NASS, L.L.; VALOIS, A.C.C. (Ed). Recursos Genéticos e Melhoramento - Microorganismos. Jaguariúna: Embrapa Meio Ambiente, Jaguariúna, 2002. p.393-407.

BANSAL, N.Production of cellulases from Aspergillus niger NS-2 in solid state fermentation on agricultural and kitchen waste residues. Waste Management, V.32, n.7, p.1341–1346, 2012.

BHAT, M. K. Cellulases and related enzymes in biotechnology. Biotechnology Advances, v.18, p.355-383, 2000.

CAMASSOLA M. & DILLON, A. J. P. Steam-Exploded Sugar Cane Bagasse for On-Site Production of Cellulases and Xylanases by Penicillium echinulatum. Energy Fuels, v.26, n.8, p. 5316–5320, 2012. Disponível em:< http://pubs.acs.org/doi/abs/10.1021/ef3009162>

CAMASSOLA M. & DILLON, A. J. P. Effect of Different Pretreatment of Sugar Cane Bagasse on Cellulase and Xylanases Production by the Mutant Penicillium echinulatum 9A02S1 Grown in Submerged Culture. BioMed Research International, v. 2014, p.1-10, 2014. Disponível em < http://dx.doi.org/10.1155/2014/720740>

CHAVES V.H. Características fisiológicas de um fungo termófilo isolado de compostagem e propriedade do seu complexo. 1982, 60f. Dissertação (Mestrado) - Universidade Federal de Viçosa, Viçosa, 1982.

CHEN, H.Z. et al. CelF of Orpinomyces PC-2 has an intron and encodes a cellulase (CelF) containing a carbohydrate-binding module. Applied Biochemistry and Biotechnology, v.108, n.1-3, p.775-786, 2003. Disponível em:< http://www.ncbi.nlm.nih.gov/pubmed/12721415>

CUNHA, F.M.; KREKE, T.; BADINO, C.S.; XIMENES, E.; LADISCH, M.R. Liquefaction of sugarcane bagasse for enzyme production. Bioresour Technol., v. 172, p. 249–252, 2014. Disponível em:

< http://www.ncbi.nlm.nih.gov/pubmed/25265329>

DING, S., G.E., W.; BUSWELL, J.A. Secretion, purification and characterization of a recombinant Volvariella volvacea endoglucanase expressed in the yeast Pichia pastoris. Enzyme and Microbial Technology, v.31, p.621-626, 2002. Disponível em:< http://www.sciencedirect.com/science/article/pii/S0141022902001680>.

GUSAKOV A.V. & SINITSYN, A.P. Cellulases from Penicillium species for producing fuels from biomass. Biofuels, v.3, n.4, p. 463-477, 2012. Disponível em: < http://www.tandfonline.com/doi/abs/10.4155/bfs.12.41?journalCode=tbfu20#preview>

HA, S.J. et al. Engineered Saccharomyces cerevisiae capable of simultaneous cellobiose and xylose fermentation. Proceedings of the National Academy of Sciences, v.108, p.504-509, 2011. Disponível em: < http://www.pnas.org/content/108/2/504.full>.

HEDGER, J.N. et al. ‘Witches brooms’ and cocoa. Journal Agricultural Society, v.69, p.164-183, 1989.

HECK, J.X. et al. Cellulase and xylanase production by isolated Amazon Bacillus strains using soybean industrial residue based solid-state cultivation. Brazilian Journal of Microbiology , v.33, p.213-218, 2002. Disponível em: <http://www.scielo.br/scielo.php?pid=S1517-83822002000300005&script=sci_arttext>

HYEON, J.E. et al. Production of minicellulosomes from Clostridium cellulovorans for the fermentation of cellulosic ethanol using engineered recombinant Saccharomyces cerevisiae. FEMS Microbiology Letters, v.310, n.1, p.39-47, 2010. Disponível em:< http://www.ncbi.nlm.nih.gov/pubmed/20637040>.

JIANG, X. et al. New isolate of Trichoderma viride strain for enhanced cellulolytic enzyme complex production. Journal of Bioscience and Bioengineering, v.11, n.2, 121-127, 2011. Disponível em:< http://www.ncbi.nlm.nih.gov/pubmed/21071269>

JÚNIOR, J.A.S.P. et al. Cellulase activity of a Lentinula edodes (Berk.) Pegl. strain grow in media containing carboximetilcellulose or microcrystalline cellulose. Brazilian Archives of Biology and Thechnology, v.46, n.3, p. 333-337, 2003. Disponível em: < http://www.scielo.br/scielo.php?pid=S1516-89132003000300004&script=sci_arttext>.

LI, X. L. et al. Properties of a recombinant beta-glucosidase from polycentric anaerobic fungus orpinomyces PC-2 and its application for cellulose hydrolysis. Applied Biochemistry and Biotechnology, v.113, p.233-250, 2004. Disponível em:< http://www.ncbi.nlm.nih.gov/pubmed/15054209>.

LI et al, 2014. Isolation, characterization and application of a cellulose-degrading strain Neurospora crassa S1from oil palm empty fruit bunch. Microbial Cell Factories, v13, p. 157-164, 2014. Disponível em:< http://www.ncbi.nlm.nih.gov/pubmed/25384340>.

LUO, H. et al. Gene cloning and expression of a new acidic family 7 endo-beta-1,3-1,4-glucanase from the acidophilic fungus Bispora sp. MEY-1. Applied Microbiology and Biotechnology, v.85. n.4, p.1015-1023, 2010. Disponível em:< http://www.ncbi.nlm.nih.gov/pubmed/19590866>

LUZ, E.D.M.N. et al. Doenças do cacaueiro. In: ZAMBOLIM L.; VALE, F.X.R. (Eds.) Controle de doenças de plantas: grandes culturas. Viçosa: Imprensa Universitária; Viçosa, 1997. p.611-655.

NARRA M. et al. Production of cellulases by solid state fermentation with Aspergillus terreus and enzymatic hydrolysis of mild alkali-treated rice straw. Bioresource Technology, v.121, p. 355-361, 2012. Disponível em:< http://www.ncbi.nlm.nih.gov/pubmed/22864171>.

PEREIRA, J.L. et al. Primeira ocorrência de vassoura-de-bruxa na principal região produtora de cacau do Brasil. Agrotópica, v.1, p.79-81, 1989.

PURDY, L. H.; SCHMIDT, R. A. Status of cacao witches' broom: biology, epidemiology and management. Annual Review Phytopathology, v.34, p.573-594, 1996. Disponível em:< http://www.ncbi.nlm.nih.gov/pubmed/15012557>.

SIGOILLOT, C. et al. Lignocellulotytic and hemicellulolytic system of Pycnoporus cinnabarinus: isolation and characterization of a cellobiose dehydrogenase and a new xylanase. Enzyme and Microbial Technology, v.31, p.876-883, 2002. Dispinível em:

http://wwwsciencedirectcom.ez278.periodicos.capes.gov.br/science?_ob=ArticleListURL&_method=list&_ArticleListID=701497244&_sort=r&_st=13&view=c&md5=e9e6e0cf0b5e0276e3bdddbab1deb888&searchtype=a>

SIQUEIRA, F.G. et al. The potential of agroindustrial residues for production of holocellulase from filamentous fungi. International Biodeterioration e Biodegradation, v.64, n.1, p.20-26, 2010. Disponível em:< http://www.sciencedirect.com/science/article/pii/S0964830509001760>.

TAKASHIMA, S. et al. Overproduction of recombinant Trichoderma reesei cellulases by Aspergillus oryzae and their enzymatic properties. Journal of Biotechnology, v.65, n. 2-3, p.163-171, 1998. Disponível em:

<http://ac.elscdn.com.ez278.periodicos.capes.gov.br/S0168165698000844/1s2.0S0168165698000844main.pdf?_tid=d031dab686dc11e4ba2c00000aacb35e&acdnat=1418924547_86d7d4a4f35b2a80df203b3cf3698119 >

ZHANG, Y.H.P. et al. Outlook for cellulase improvement: screening and selection strategies. Biotechnology Advances, v.24, p.452‑481, 2006. Disponível em:< www.elsevier.com/locate/biotechadv>.

ZHANG, L. et al. Effects of acid and alkali treated lignocellulosic materials on cellulase/xylanase production by Trichoderma reesei Rut C-30 and corresponding enzymatic hydrolysis. Biomass and Bioenergy, v.37, p.16-24, 2012. Disponível em:< http://www.elsevier.com/locate/biombioe>

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Published

2017-06-12

How to Cite

Guedes, C. E. S., Alvarez, R. C., Ximenes, E. A., Paulillo, L. C., Lima, S. T. C., & Barbosa, L. V. (2017). Enzyme activity of cellulases produced by Moniliophtora perniciosa, the causing factor of “witch–broom” in cacao plants. MAGISTRA, 26(4), 545 – 553. Retrieved from https://www3.ufrb.edu.br/index.php/magistra/article/view/4010

Issue

Vol. 26 No. 4 (2014)

Section

Artigo Científico

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