Ifa Manzila, Tri Puji Priyatno, Muhammad Faris Fathin, Laksmi Ambarsari, Yadi Suryadi, I Made Samudera, Dwi Ningsih Susilowati
| Abstract views: 319 | PDF views: 416


Pathogenic fungus is one of the constraints to increase crop production. Chemical control using fungicides caused negative effects either to the environment or increased pathogen resistance to fungicide. Biological control using microbial-producing ß glucanase is an alternative method to inhibit the growth of pathogenic fungus. The aim of this study was to characterize ß-1,3-1,4-glucanase produced by rice endophytic bacterium, B. cepacia E76. Purification was carried out by ammonium sulphate precipitation, dialysis, and ion exchange chromatography using DEAE sepharose Fast Flow. A further characteristic of the enzyme activity was studied using oatmeal-glucan substrate.Results showed that precipitation using saturated 80% ammonium sulphate generated a good yield with the purity increased by 11 fold and yield of 66%.After chromatography step, the ß-1,3-1,4-glucanase of B. cepacia was successfully purified with an increasedof purity up to 33 fold and yield of 4%. Based on 10% SDS-PAGE, the enzyme profiles had the molecular weight of 15, 48 and 55 kDa.Of the three isozymes, only the 48 kDa isozyme showed the strongest glucanase activity when grown on media containing glucan as substrate.


Burkholderia cepacia E76, ß-1,3-1,4-glucanase, purification, rice

Full Text:



Adams DJ. 2004. Fungal cell wall chitinases and glucanases. Microbiology 150, 2029-2035.

Aono R, M Hammura, M Yamamoto and T Asano. 1995. Isolation of extracellular 28 and 42 kilodalton ?-1,3-glucanases and comparison of three ?-1,3-glucanases produced by Bacillus circulans IAM1165. Applied Environmental Microbiology 61, 122-129.

Bayer EA, J Belaich, Y Shoham and R Lamed. 2004. The cellulosomes: multienzyme machine for degradation of plant cell wall polysaccharides. Annual Reviewof Microbiology 58, 521-554.

Bevivino A, C Dalmastri, STabacchioni and L Chiarini. 2000. Efficacy of Burkholderia cepacia MCI 7 in disease suppression and growth promotion of maize. Biology and Fertility of Soils Journal 31(3-4), 225-231.

Bradford MM. 1976. A rapid and sensitive method of microgram quantities of protein utilizing the principle of proteindye binding. Analytical Biochemistry 12, 248-254.

Blazina M, M Najdek, A Ruso and D Fuks. 2010. Adaptational changes in cellular fatty acids of cultured bacteria as a response to tropic differences. Acta Adriatica 51,131-140.

Bollag DM and SJ Edelstein. 1991. Protein Methods, 415.Wiley Liss, New York.

Celestino KRS, RB Cunha and CR Felix. 2006. Characterization of a ?-glucanase produced by Rhizopus microsporus var.microsporus and its potential for application in the brewing industry. BioMed Central Biochemistry 7, 23-31.

Celestino KRS, RB Cunha and CR Felix. 2006. Characterization of a ?-glucanase produced by Rhizopus microsporus var.microsporus and its potential for application in the brewing industry. BioMed Central Biochemistry 7, 23-31.

Chen JP and MS Lee. 1994. Simultaneous production and partition of chitinase during growth of Serratiamarcescens in an aqueous two-phase system. Biotechnoogy Techniques 8(11), 783-788.

Doxey AC, MW Yaish, BA Moffatt, M Griffith and BJ McConkey. 2007. Functional divergence in the Ara-bidopsis ?-1,3-glucanase gene family inferred by phylogenetic reconstruction of expression state. Molecular Biology and Evolution 24, 1045-1055.

Erfle JD and RM Teather. 1991. Isolation and properties of a (1,3)-?-D-glukanase from Ruminococcus flavefaciens. Applied Environmental Microbiology 57, 122-129.

Heungens K and JL Parke. 2000. Zoospore homing and infection events: effects of the biocontrol bacterium Burkholderia cepacia AMMDR1 on two oomycete pathogen of pea (Pisum sativum). Applied Environmental Microbiology 66, 5192-5200.

Kang Y, R Carlson, W Tharpe and MA Schell. 1998. Characterization of genes involved in biosynthesis of a novel antibiotic from Burkholderia cepacia BC11 and their role in biological control of Rhizoctonia solani. Applied Envi-ronmental Microbiology 64, 3939-3947.

Kobayashi DY and JD Palumbo. 2000. Bacterial endophytes and their effects on plants and uses inagriculture. In: Microbial Endophytes. C.W. Bacon and J.F.White (eds.), 199-233. Marcel Dekker, Inc, New York.

Maheshwari DK, M Saraf and A Aeron. 2013. Bacteria in Agrobiology: Crop Ecosystems, 220-254. Springer, New York.

Mendes R, AA Pizzirani-Kleiner, WL Araujo and JM Raaijmakers. 2007. Diversity of cultivated endophytic bacteria from sugarcane: genetic and biochemical characterization of Burkholderia cepacia complex isolates. Applied Environmental Microbiology 73, 7259-7267.

Miller GL. 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry 31 (2), 426-428.

Monisha R, MV Uma and K Murthy. 2009. Partial purification and characterization of Bacilus pumilus xylanase from soil source. Kathmandu University Journal of Science, Engineering and Technology 5, 137-148.

Nelson DL and MM Cox. 2012. Lehninger Principles of Biochemistry, 1100, sixth ed. WH Freeman and Company. New York.

Nishiyama Y, S Kuga and S Sato. 2001. Role of the putative membranebound putative endo-?-D-1,4-glucanase KOR-RIGAN in cell elongation and cellulose synthesis in Arabidopsis. Plant Cell Physiology 42,251-263.

Meyer JM, V Tran, A Stinzi, O Berge and G Winkelman. 1995. Ornibactin production and transport properties in strains of Burkholderia cepacia and Burkholderia vetnamienses (formely Pseudomonas cepacia). Biometals 8, 309-307.

Shetty NP, JD Jensen, A Knudsen, C. Finnie, N Geshi, A Blennow, DB Collinge and HJL Jorgensen.2009. Effect of ?-1,3-glucan from Setoria tritici on structural defence responses in wheat. Journal of Experimental Botany 60, 4287-4300.

Suryadi Y, TP Priyatno, IM Samudra, DN Susilowati, Patricia dan W Irawati. 2013. Karakterisasi dan identifikasi isolat bakteri endofitik penghambat jamur patogen padi. Buletin Plasma Nutfah 19 (1),25-32.

Suryadi Y, DN Susilowati, P Lestari, TP Priyatno, IM Samudra, N Hikmawati and NR Mubarik. 2014. Characterization of bacterial isolates producing chitinase and glucanase for biocontrol of plant fungal pathogens. Journal of Agricultural Technology 10(4), 983-999.

Strobel GA and B Daisy. 2003. Bioprospecting for microbial endophytes an their natural products. Microbiology and Molecular Biology Review 67(4), 63-68.

Wood PJ and J Weisz. 1984. Use of calcuflour in analysis of oat beta-D-glucan. Cereal Chemistry 6,73-75.

Wolf N. 1991. Complete nucleotide sequence of Hordeum vulgare gene encoding ?-(1,3-1,4)-glucanase isoenzyme II. Plant Physiology 96, 1382-1384.

Wolf M, A Geczi, O Simon and R Borriss. 1995. Genes encoding xylan and ?-glucan hydrolising enzymes in Bacillus subtilis: characterization, mapping, and construction of strains deficient in lichenase, cellulase, xylanase. Micro-biology 141, 281-290.

Yang S, Y Qiaojuan, Z Jiang, G Fan and L Wang. 2008. Biochemical characterization of a novel thermostable ?-1,3-1,4-glucanase from Paecilomyces thermophila. Journal of Agriculture and Food Chemistry 56, 5345-5351.

Zeng YF, JH Ying, JC Ming, CP Chi, JTC Tzen and RL Je. 2009. Simultaneous refolding, purification, and immobilization of recombinant Fibrobacter succinogenes 1,3-1,4-?-glucanase on artificial oil bodies. Journal of Chemical Technology and Biotechnology 84, 1480-1485.

Zverlov VV, K Fuchs andWH Scwarz. 2002. Chi18A, the endochitinase in the cellulosome of the thermophilic cellulolytic bacterium Clostridium thermocellum.Applied and Environmental Microbiology 68, 3176-3179.


  • There are currently no refbacks.