The rising concerns about the scarcity of fossil fuels, the emission of green house gasses and air pollution by incomplete combustion of fossil fuel have also resulted in an increasing focus on the use of cellulases to perform enzymatic hydrolysis of the lignocellulosic materials for the generation of bioethanol. The aim of this study was to isolate a potential thermo-solvent tolerant cellulase producing bacterium from natural resources, and then applied for purification and characterization. The purified enzyme was to be accessible for the bioethanol production as well as industrial exploitation (discuss in our next study). It is the first instance when thermo-solvent tolerant cellulase producing bacterium was isolated from soil sample. The culture was identified as Bacillus vallismortis RG-07 by 16S rDNA sequence analysis. Bacillus vallismortis RG-07 reported maximum cellulase production from sugarcane baggase (4105 U ml−1) used as agro-waste carbon source. The cellulase enzyme produced by the Bacillus sp. was purified by (NH4)2SO4 precipitation, ion exchange and gel filtration chromatography, with overall recovery of 28.8%. The molecular weight of purified cellulase was 80 kDa as revealed by SDS-PAGE and activity gel analysis. The optimum temperature and pH for enzyme activity was determined as 65°C and 7.0 and it retained 95 and 75% of activity even at 95°C, and 9.0 respectively. The enzyme activity was enhanced in the presence of organic solvents (30%) n-dodecane, iso-octane, n-decane, xylene, toluene, n-haxane, n-butanol, and cyclohexane, after prolonged incubation (7 days). The enzyme activity was also stimulated by Ca2+, mercaptoethanol, Tween-60, and Sodium hypochloride whereas strongly inhibited by Hg. Kinetic analysis of purified enzyme showed the Km and Vmax to be 1.923 mg ml−1 and 769.230 μg ml−1 min−1, respectively. The unique property of solvent-thermostable-alkalophilic, nature proves the potential candidature of this isolate for current mainstream biomass conversion into fuel and other industrial process.

Aureobasidium pullulans, popularly known as black yeast, is one of the most widespread saprophyte fungus associated with wide range of terrestrial and aquatic habitats, in temperate and tropical environment. It is a polymorphic fungus that is able to grow in single yeast-like cells or as septate, polykaryotic hyphae showing synchronous conditions, with budding cells. This fungus has been exploited potentially for commercial production of various enzymes (amylase, xylanase, pectinase, etc). Single cell protein, alkaloids and polysaccharide, especially pullulan, an exopolysaccharide, is a linear α-d-glucan connected with α-1,4 glycosidic bond mainly of maltotriose repeating units interconnected by α-1,6 linkages. Pullulan has been considered as one of the important polysaccharide for production of biodegradable plastics. More than 300 patents for applications have been developed. It is the only fungus which produce higher amount of pullulan and has been exploited all over the world. The fungus has excellent genetic make-up to produce various important metabolites at commercial production with limited species. Some of the A. pullulans have potential antagonistic activity against a number of phytopathogenic fungi used as bio-control agents of post-harvest diseases. It has been found to be tolerant to many metal ions which are common pollutants of soil and water. Several strains of this fungus have ability to degrade xenobiotic compounds. In the light of the above facts, this review article has emphasized on the orientation, morphology, biochemical characteristics, habitats and its economic potentials with special reference to pullulan. Key words: Aureobasidium pullulans, single cell protein, xenobiotic compounds, pullulan.

Total 95 isolates of Aureobasidium pullulans were isolated from different flowers and leaves samples, out of which 11 thermotolerant strains produced pullulan. One thermotolerant non-melanin pullulan producing strain, designated as RG-5, produced highest pullulan (37.1±1.0 g/l) at 42ºC, pH 5.5 in 48h of incubation with 3% sucrose and 0.5% ammonium sulphate in a non-stirred fed batch fermentor of 6 liters capacity. The two liters of initial volume of fermentation medium was further fed with the 2 liters in two successive batches at 5 h interval into the fermentor. The sterile air was supplied only for 10h at the rate of 0.5 vvm.

Sixty bacterial strains isolated from the soils sample in the presence of organic solvent were screened for xylanase production. Among them, strain RSPP-15 showed the highest xylanase activity which was identified asBacillus vallismortis. The isolate showed maximum xylanase production (3768 U/mL) in the presence of birch wood xylan and beef extract at 55°C pH 7.0 within 48 h of incubation. The enzyme activity and stability were increased 181.5, 153.7, 147.2, 133.6, and 127.9% and 138.2, 119.3, 113.9, 109, and 104.5% in the presence of Co²⁺, Ca²⁺, Mg⁺², Zn⁺², and Fe⁺³ions (10 mM). Xylanase activity and stability were strongly inhibited in the presence of Hg and Cu ions. The enzyme was also stable in the presence of 30% ofn-dodecane, isooctane,n-decane, xylene, toluene,n-hexane,n-butanol, and cyclohexane, respectively. The presence of benzene, methanol, and ethanol marginally reduced the xylanase stability, respectively. This isolate may be useful in several industrial applications owing to its thermotolerant and organic solvent resistance characteristics.

In the present study, an extracellular alkali-thermo-tolerant xylanase from Bacillus paramycoides was produced in the presence of an organic solvent. The enzyme was purified by ammonium sulphate precipitation, gel filtration, and ion exchange chromatography, with an overall recovery of 25.9%. The purified enzyme hada 70 kDa molecular weight (MW) confirmed by SDS-PAGE gel analysis. The maximum enzyme activity was reported at 55 °C and pH 7.0. Xylanase activity and stability were improved in the presence of 30% (v/v) n-dodecane, iso-octane, n-decane, and cyclohexane (7 days). The enzyme activity was improved by Co²⁺, EDTA, and Triton-X-100 while vigorously repressed by Hg²⁺ and Cu²⁺. The purified enzyme showed 1.473 mg/mL K_m and 654.017 µg/mL/min V_max values. The distinctive assets of the isolate verified the potential application in the field of biomass conversion into fuel and other industrial processes. Organic solvent-tolerant xylanases can be used for concurrent saccharification and bioethanol production, the amplification of intoxicating beverages, and the fermenting industry.