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Screening of Cellulase, Pectinase and Xylanase Activities and Optimization of Radial Mycelial Growth of Two Thermophilic Fungi

KUMKUM AZAD
Department of Botany, Jahangirnagar University, Savar, Dhaka-1342, Banglades

FEROZA HOSSAIN
Department of Botany, Jahangirnagar University, Savar, Dhaka-1342, Banglades

MD ABDUL HALIM
Department of Botany, Jahangirnagar University, Savar, Dhaka-1342, Banglades

Abstract/ Executive Summary:

The enzymatic activity of Thermomyces lanuginosus BPJ-10 and Rhizomucor pusillus BPJ-2 were observed through qualitative screening programme which was demonstrated by the hydrolysis of substrate on solid media. Both the fungi exhibited potential xylanolytic and pectinolytic activities whereas no cellulase activity was observed in T. lanuginosus BPJ-10 and very low cellulase activity was found in R. pusillus BPJ- 2. The optimum temperature for mycelial growth of T. lanuginosus BPJ-10 and R. pusillus BPJ-2 were found to be 50 and 45°C, respectively, whereas the optimum pH for both of them were 6.5 and 5, respectively. Out of five culture media used, both the fungi showed maximum radial mycelia growth on Potato Dextrose Agar.

Keywords: Thermophilic fungi, Cellulase, Polygalacturonase, Xylanase,Thermomyces lanuginosus, Rhizomucor pusillus

Location: Jahangirnagar University, Savar, Dhaka

Start Date:

End Date:

Program Area: Pest Management

Commodity: Diseases

Objectives:

To study enzymatic activity and radial mycelial growth.

Methodology:

Saw dust, piles of hay and soil were used for isolation of thermophilic fungi. Samples were inoculated on PDA medium (pH 6.5) and incubated at 50ºC temperature. After 4 days of incubation, different colonies with different colours and shapes were found on the culture plates. Then distinct colonies were isolated on PDA medium as pure culture. Pure fungal mycelium was transferred to agar slant for stock culture and the culture was maintained at 4°C and subcultured at 14 days intervals. Then the colonies were observed under microscope. Enzyme activity to be demonstrated by the hydrolysis of substrate incorporated, generally as the main carbon source such as carboxymethyle cellulose (CMC) for carboxymethyle cellulase (CMCase) activity, xylan for xylanase activity and polygalacturonic acid (PG) for polygalacturonase (PGase) activity, respectively on a solid agar medium following the method of Mohiuddin (1992). For screening of CMCase activity the fungi were grown on CMC-agar. Culture plates of 4 days were flooded with 10 ml Congo red (0.1%) solution. After 20 minutes, the dye was replaced by 5 mol/l NaCl solution and CMCase activity was revealed by a pale orange zone around the colonies. For the screening of PGase activity, the fungi were grown on PG-agar. After 5 days of growth the plates were flooded with 1% (w/v) cetyltrimethyl ammonium bromide solution. PGase activity was observed by the formation of hydrolytic zone around the colonies after 5 - 10 min of incubation. For screening of xylanase activity fungal colonies were grown on xylan-agar media. After 4 days the plates were flooded with 96% ethanol. Xylanase activity exhibited by the formation of hydrolytic zone around the colonies after 3 - 4 hrs of incubation. To determine the maximum vegetative growth, the fungal isolates were allowed to grow on PDA medium at different temperatures such as 40, 45, 50, 55 and 60°C. To determine the optimum pH for maximum mycelial growth, the fungal isolates were cultured on PDA mediun at pH 4, 4.5, 5, 5.5, 6, 6.5, 7, 8 and 9. To investigate the effect of different culture media on radial mycelial growth, the isolated fungi were allowed to grow on different solid media such as carboxy methyl cellulose (CMC), Czap ek’s Dox Agar, Malt Extract Ag ar (MEA), Mendel’s and PDA media (Hossain et al. 1999 and Mohiuddin 1992). For each case radial mycelial growth of fungi was recorded at every 24 hrs up to 7 days. Data obtained from radial mycelial growth diameters were analysed statistically. Means were compared by least significant di fference (LSD) at 5% level of significance through one way ANOVA and DMRT by using SPSS program 11.

Source of Information: Bangladesh J. Bot. 42(2): 207-213, 2013 (December

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Both the fungi exhibited potential xylanolytic and pectinolytic activities whereas no cellulase activity was observed in T. lanuginosus BPJ-10 and very low cellulase activity was found in R. pusillus BPJ- 2. The optimum temperature for mycelial growth of T. lanuginosus BPJ-10 and R. pusillus BPJ-2 were found to be 50 and 45°C, respectively, whereas the optimum pH for both of them were 6.5 and 5, respectively. Out of five culture media used, both the fungi showed maximum radial mycelia growth on Potato Dextrose Agar.

Knowledge Management: Journal