Amyloid protein produced by B. cereus CR4 possesses bioflocculant activity and has potential application in microalgae harvest.

Bacillus cereus CR4 from the flocs of activated sludge was found to produce an extracellular bioflocculant, which was characterized as amyloid protein and demonstrated to have potential application in microalgae recovery. Cell surface amyloid production was demonstrated by fluorescence, confocal and scanning electron microscopy. Birefringence, spectral shift assay, TEM, FTIR and CD spectra confirmed the amyloid nature of the purified protein that demonstrated flocculation. The gene for amyloid protein of B. cereus CR4 was found to be related to tasA gene of amyloid protein produced by Bacillus subtilis. The results demonstrated that the amyloid protein produced by B. cereus CR4 possessed a novel bioflocculant activity which at pH below 4.5 reached to a maximum of 86.87%. The amyloid bioflocculant producing B. cereus CR4 has a potential in biotechnological application like Scenedesmus biomass recovery.

Physicochemical characterization of pectinase activity from Bacillus spp. and their accessory role in synergism with crude xylanase and commercial cellulase in enzyme cocktail mediated saccharification of agrowaste biomass.

AIM: The aim of this study was to evaluate the physicochemical properties of the crude pectinase activity from three Bacillus isolates of ruminant dung origin and study their synergism with crude xylanases from the same Bacillus spp. and a commercial cellulase to evaluate their accessory role in improved biomass saccharification. METHODS AND RESULTS: Pectinolytic crude culture filtrate obtained from three ruminant dung isolates, Bacillus safensis M35, Bacillus altitudinis R31 and Bacillus altitudinis J208, on crude pectin containing medium possessed polygalacturonate hydrolase, pectate lyase and pectin lyase activities. Studies regarding their stability under various temperature and pH conditions revealed their mild acidic to alkaline and mesophilic nature with enzyme activity falling within the pH range 6·0-9·0 and temperature range 30-60°C. The pectinase activity was categorized as endolytic as it brought about ~50% reduction in relative viscosity of pectic polymer within initial 10 min of incubation. Synergism of pectinase activity with crude xylanase activities and/or commercial cellulase was clearly demonstrated as ~1·6 to ~1·9-fold increase in agrowaste biomass saccharification was obtained confirming the role of pectinases as accessory enzymes. CONCLUSION: Synergism of the broad-spectrum endopectinase activity obtained from three Bacillus isolates with accessory crude xylanases from the same isolates and commercial cellulase enhanced the agrowaste saccharification and confirmed the accessory role of crude pectinase as they formed an efficient enzyme cocktail functioning in a contributive manner for improvement of agrowaste biomass saccharification. SIGNIFICANCE AND IMPACT OF THE STUDY: Mesophilic crude endopectinases obtained from Bacillus spp. isolated from ruminant dung possessed activity in broad pH and temperature ranges as well as broad substrate specificity. Moreover, their synergism with crude xylanase and Primfast® 200 cellulase demonstrated the potential to form efficient enzyme cocktail for application in plant biomass saccharification process.

Bioemulsifier (BE-AM1) produced by Solibacillus silvestris AM1 is a functional amyloid that modulates bacterial cell-surface properties.

A novel estuarine bacterial strain, Solibacillus silvestris AM1, produces an extracellular, thermostable and fibrous, glycoprotein bioemulsifier (BE-AM1). The amyloid nature of the bioemulsifier (BE-AM1) was confirmed by biophysical techniques (Congo red based polarization microscopy, ThioflavinS based fluorescent microscopy, fibrous arrangement in transmission electron microscopy and secondary structure measurement by FTIR and CD spectrum analysis). Cell-bound BE-AM1 production by S. silvestris AM1 during the mid-logarithmic phase of growth coincided with a decrease in cell surface hydrophobicity, and an increase in cell autoaggregation and biofilm formation. It was observed that the total interfacial interaction energy ([Formula: see text]) for the surface of the bioemulsifier producing S. silvestris AM1 and different derivatized surfaces of polystyrene (silanized and sulfonated) was found to support biofilm formation. This study has revealed that the BE-AM1, a bacterial bioemulsifier, is a functional amyloid and has a role in biofilm formation and cell surface modulation in S. silvestris AM1.

Microcosm-based interaction studies between members of two ecophysiological groups of bioemulsifier producer and a hydrocarbon degrader from the Indian intertidal zone.

Isolates were obtained from intertidal zone site samples from all five western and one eastern coastal states of India and were screened. These ecophysiological groups of aerobic, mesophilic, heterotrophic, sporulating, and bioemulsifier-producing bacteria were from Planococcaceae and Bacillaceae. This is the first report of bioemulsifier production by Sporosarcina spp., Lysinibacillus spp., B. thuringiensis, and B. flexus. In this group, Solibacillus silvestris AM1 was found to produce the highest emulsification activity (62.5 %EI) and the sample that yielded it was used to isolate the ecophysiological group of non-bioemulsifier-producing, hydrocarbon-degrading bacteria (belonging to Chromatiales and Bacillales). These yielded hitherto unreported degrader, Rheinheimera sp. CO6 which was selected for the interaction studies (in a microcosm) with bioemulsifier-producing S. silvestris AM1. The gas chromatographic study of these microcosm experiments revealed increased degradation of benzene, toluene, and xylene (BTX) and the growth of Rheinheimera sp. CO6 in the presence of bioemulsifier produced by S. silvestris AM1. Enhancement of the growth of S. silvestris AM1 in the presence of Rheinheimera sp. CO6 was observed possibly due to reduced toxicity of BTX suggesting mutualistic association between the two. This study elucidates the presence and interaction between enhancers and degraders in a hydrocarbon-contaminated intertidal zone and contributes to the knowledge during application of the two in remediation processes.

Xylanases of Bacillus spp. isolated from ruminant dung as potential accessory enzymes for agro-waste saccharification.

UNLABELLED: Polysaccharide hydrolase producing bacteria were isolated for biomass saccharification step in two-step bioethanol production. Xylanolytic bacteria were found to be common in ruminant dung. Seven Bacillus dung isolates exhibited high xylanolytic activity; three of which were identified as Bacillus safensis and four as Bacillus altitudinis, based on 16S rDNA and gyrB gene sequencing. Interestingly, comparison of activity profiles for B. safensis M35 and B. altitudinis R31 and J208 crude xylanases showed activity in similar temperature and pH ranges of 40-60 °C and 6·0-9·0, respectively, even though they were isolated from three different dung sources. Furthermore, 22-28% viscosity reduction of beechwood xylan substrate by all the three xylanases points towards their endo-acting nature. Endo-acting xylanases are envisaged as accessory enzymes which help expose the cellulose fibres for the subsequent action of the core enzyme cellulases. In this study, when supplemented to the commercial cellulase as a cocktail, the accessory role of the crude xylanases from the selected Bacillus strains was established as 1·3, 2·33 and 1·9 fold increase in saccharification of barley husk, sugarcane bagasse and wheat husk was achieved, respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: The uncontrolled use of fossil fuels and concerns about its future availability, have invoked interest over unconventional alternative energy sources like solar, hydropower, geothermal, nuclear and biomass. Plants, being largest renewable biomass on earth, have received consideration as a source of biofuels. Ruminant dung isolates M35, R31 and J208 belonging to Bacillus sp. produces majorly endo-xylanase when induced with wheat bran. Such plant cell wall degrading endo-xylanases with broad pH optima and mesophilic nature can act as accessory enzymes with cellulases to enhance the saccharification of plant biomass in biofuel industries.

Nutrients determine the spatial architecture of Paracoccus sp. biofilm.

Bacterial biofilms adapt and shape their structure in response to varied environmental conditions. A statistical methodology was adopted in this study to empirically investigate the influence of nutrients on biofilm structural parameters deduced from confocal scanning laser microscope images of Paracoccus sp.W1b, a denitrifying bacterium. High concentrations of succinate, Mg(++), Ca(++), and Mn(++) were shown to enhance biofilm formation whereas higher concentration of iron decreased biofilm formation. Biofilm formed at high succinate was uneven with high surface to biovolume ratio. Higher Mg(++) or Ca(++) concentrations induced cohesion of biofilm cells, but contrasting biofilm architectures were detected. Biofilm with subpopulation of pillar-like protruding cells was distributed on a mosaic form of monolayer cells in medium with 10 mM Mg(++). 10 mM Ca(++) induced a dense confluent biofilm. Denitrification activity was significantly increased in the Mg(++)- and Ca(++)-induced biofilms. Chelator treatment of various biofilm ages indicated that divalent cations are important in the initial stages of biofilm formation.

Alcaligenes faecalis kw-a biofilm for denitrification of nitrate-rich effluent.

Alcaligenes faecalis kw-A selected for possessing good denitrification efficiency was used for biofilm development. The biofilm could be developed on a glass surface within 12 hr when 5%, Ix 10(8) cells/ml was used as inoculum. The microcolonies were seen in 6 hr and glycocalyx in 9 hr stage. At 24 hr the biofilm was developed fully and hence was visualised as dense mass. The biofilm protein content showed 48.5% increase in shake flask than in static condition. The exopoplymer is produced in larger amounts in biofilm as compared to the suspended cells. Also, its amount was more by 43% in the biofilm produced in shake flask condition than in static condition. The biofilm could remove 95% nitrate from nitrate-rich effluent in a bench-scale process in 36 hr. The attached growth technique demonstrated here can be utilised to study the effect of favourable as well as adverse conditions on the denitrification efficiency of a culture. The ultimate application of a denitrifying biofilm would be in attached growth or biofilm reactor.

Septic abortion.

PIP: 53 of 3100 abortions at Bombay hospital were septic abortions, giving an incidence of 1.7%. Various factors of possible etiological significance were analyzed, including age, parity, marital status, duration of gestation, and the mode of interference leading to sepsis. 36 of 53 patients were aged 20-30 years, but other age groups were represented. In the present study, gravidity was not relevant, for all gravidity groups, from primipara to grand multipara 5 and above, had patients suffering septic abortions. 9 patients were married and gave a history of interference; in all, 38 patients were married, 22 were unmarried, and 4 were widows. 23 patients gave a definite mode of interference, and the most common method was interference with a stick. 43% mortality occurred in patients giving a history of interference, and 36% mortality occurred in others. Vaginal and cervical cultures revealed (16 cases studied) 5 cases of CL. tetani, 1 case of E. coli, and 10 patients showing strepto-, staphylo-, pneumococcal infections. In this series, 21 of 53 patients died: 8 of tetanus, 3 or renal failure, 4 of septicemia, 2 of hemorrhagic diathesis, and 3 of endotoxic shock. 1 patient had acute bacterial endocarditis and pulmonary embolism at sutopsy. It is this article's contention that the main cause of sepsis is using an instrument to induce abortion during an unwanted pregnancy; hence, a plea is made for more liberalized abortion legislation.^ieng