Studies on Production of Bacterial Xylanases

Xylanases with hydrolytic activity on xylan, one of the hemicellulosic materials present in plant cell walls, have been identified long back and the applicability of this enzyme is constantly growing. All these applications especially the pulp and paper industries require novel enzymes. There has been lot of documentation on microbial xylanases, however, none meeting all the required characteristics. The characters being sought are: higher production, higher pH and temperature optima, good stabilities under these conditions and finally the low associated cellulase and protease production. The present study analyses various facets of xylanase biotechnology giving emphasis on bacterial xylanases. Fungal xylanases are having problems like low pH values for both enzyme activity and growth. Moreover, the associated production of cellulases at significant levels make fungal xylanases less suitable for application in paper and pulp industries.Bacillus SSP-34 selected from 200 isolates was clearly having xylan catabolizing nature distinct from earlier reports. The stabilities at higher temperatures and pH values along with the optimum conditions for pH and temperature is rendering Bacillus SSP-34 xylanase more suitable than many of the previous reports for application in pulp and paper industries.Bacillus SSP-34 is an alkalophilic thertmotolerant bacteria which under optimal cultural conditions as mentioned earlier, can produce 2.5 times more xylanase than the basal medium.The 0.5% xylan concentration in the medium was found to the best carbon source resulting in 366 IU/ml of xylanase activity. This induction was subjected to catabolite repression by glucose. Xylose was a good inducer for xylanase production. The combination of yeast extract and peptone selected from several nitrogen sources resulted in the highest enzyme production (379+-0.2 IU/ml) at the optimum final concentration of 0.5%. All the cultural and nutritional parameters were compiled and comparative study showed that the modified medium resulted in xylanase activity of 506 IU/ml, 5 folds higher than the basal medium.The novel combination of purification techniques like ultrafiltraton, ammonium sulphate fractionation, DEAE Sepharose anion exchange chromatography, CM Sephadex cation exchange chromatography and Gel permeation chromatography resulted in the purified xylanase having a specific activity of 1723 U/mg protein with 33.3% yield. The enzyme was having a molecular weight of 20-22 kDa. The Km of the purified xylanase was 6.5 mg of oat spelts xylan per ml and Vmax 1233 µ mol/min/mg protein.Bacillus SSP-34 xylanase resulted in the ISO brightness increase from 41.1% to 48.5%. The hydrolytic nature of the xylanase was in the endo-form.Thus the organism Bacillus SSP-34 was having interesting biotechnological and physiological aspects. The SSP-34 xylanase having desired characters seems to be suited for application in paper and pulp industries.

Biogeochemistry of the shelf sediments of south eastern Arabian sea: Effect on benthic bacterial heterotrophs

The composition and variability of heterotrophic bacteria along the shelf sediments of south west coast of India and its relationship with the sediment biogeochemistry was investigated. The bacterial abundance ranged from 1.12 x 103 – 1.88 x 106 CFU g-1 dry wt. of sediment. The population showed significant positive correlation with silt (r = 0.529, p< 0.05), organic carbon (OC) (r = 0.679, p< 0.05), total nitrogen (TN) (r = 0.638, p< 0.05), total protein (TPRT) (r = 0.615, p< 0.05) and total carbohydrate (TCHO) (r = 0.675, p< 0.05) and significant negative correlation with sand (r = -0.488, p< 0.05). Community was mainly composed of Bacillus, Alteromonas, Vibrio, Coryneforms, Micrococcus, Planococcus, Staphylococcus, Moraxella, Alcaligenes, Enterobacteriaceae, Pseudomonas, Acinetobacter, Flavobacterium and Aeromonas. BIOENV analysis explained the best possible environmental parameters i.e., carbohydrate, total nitrogen, temperature, pH and sand at 50m depth and organic matter, BPC, protein, lipid and temperature at 200m depth controlling the distribution pattern of heterotrophic bacterial population in shelf sediments. The Principal Component Analysis (PCA) of the environmental variables showed that the first and second principal component accounted for 65% and 30.6% of the data variance respectively. Canonical Correspondence Analysis (CCA) revealed a strong correspondence between bacterial distribution and environmental variables in the study area. Moreover, non-metric MDS (Multidimensional Scaling) analysis demarcated the northern and southern latitudes of the study area based on the bioavailable organic matter