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Detection and Composition of Extracellular Polisaccharide in Bacteria

M. A. Z. Molla
Department of Microbiology, University of Dhaka, Dhaka-1000, Bangladesh

A. A. Chowdhury
Department of Microbiology, University of Dhaka, Dhaka-1000, Bangladesh

A. Islam
Department of Soil Science, University of Dhaka, Dhaka-1000, Bangladesh

S. Hoque
Department of Soil Science, University of Dhaka, Dhaka-1000, Bangladesh

Abstract/ Executive Summary:

Extracellular polysaccharide producing bacteria were isolated from soil and water samples collected from different places of Dhaka city. These isolates were identified to be strain of Serratia marcescens, Pseudomonas fluorescens, Azotobacter chrococcum, Azotobacter vineiandii and Flavobacterium sp. The highest and lowest amount of polysaccharide were produced by Serratia marcescens and Pseudomonas fluorescens, respectively. The extracellular polysaccharide produced by these bacteria showed differences in chemical composition. The extracellular polysaccharide produced by an isolate identified to be strain of Serratia marcescens contained glucose, galactose, glucuronic acid, mannose and rhamnose but a strain of Pseudomonas fluorescens produced a polysaccharide which contained glucose, galactose and mannose.

Keywords: Extracellular polysaccharide, Producing bacteria, Local habitats, Potential isolates, Optimal production

Location: Department of Microbiology, University of Dhaka, Dhaka-1000, Bangladesh

Start Date:

End Date:

Program Area: Pest Management

Commodity: Insects

Objectives:

To isolate the extracellular polysaccharide producing bacteria of local habitats and the potential isolates were characterized for optimal production of extracellular polysaccharide in culture media and the individual component of the polysaccharide were also analysed.

Methodology:

The formation of mucoid layer around the colony on agar plate indicated the polysaccharide producing bacteria. The isolated colonies were identified according to the Bergey’s Manual of Determinative Bacteriology (3). the isolated colonies were maintained on Yeast mannitol agar (1) slant and Burk’s nitrogen free (2) agar slant. Bacteria were cultured in 50cmr Erlenmeyer flasks containing 100 ml and Burk's nitrogen free (2) liquid media in two replicates for each culture. The flasks were aerated on a reciprocal shaker at speed of 120 strokes/min. for 5 days (Fooetrakul et al. media containing flasks) and for 8 days (Burk's nitrogen free media containing flasks) at room temperature. The cultures diluted with an equal volume of distilled water were centrifuged at 13,000 rpm for t hour, using a superspeed centrifuge and the cell free solution was sperated from the pellet. Two volume or acetone was added to the supernatant with stirring. The precipitated polysaccharide was washed with water acetone (l : 2 by volume), dried with acetone in vacuo and weighed (1). The polysaccharide was purified by gel filtration through sephadex G-25 (Pharmacia) in a 2.5×70 cm column. 10ml of 2mg ml-1 solution of the polysaccharide in distilled water were put on the top of the column and 3ml fractions were collected at a flux of 0.5m1 min-1(4). Polysaccharide was detected by the anthrone reaction(5). Descending paper chromatography (whatman No.1) was used with n-butanol : pyridine : water (6 : 4 :.3.) as solvent. Aniline acid phthalate (930mg of aniline and 1.6gm or phthalic acid in 100ml saturated n-butanol) was used as developer for carbohydrates. All identification of spots were based on comparisons with authentic sugar samples(4).

Source of Information: Dhaka University Studies B, XXXIII, (1) : 17-21, 1985 (January).

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The results obtained in the present study was consistent with the finding of extracellular material. The unpurified extracellular material obtained from S. marcescens were rhamnose, glucose and smaller amounts of galactose, galactoseamine, glucoseamine and bound lipid. A. vinelandii contained galacturonic acid, glucose, rhamnose with various anounts of O-acyl groups and other sugars. A. chrococcum contained glucose, galactose, rhamnose and either galacturonic acid or glucuronic acid (9, 10) and another strain contained xylose.

Knowledge Management: Journal