A Computational Approach to Incorporate Metabolite Inhibition in the Growth Kinetics of Indigenous Bacterial Strain Bacillus subtilis MN372379 in the Treatment of Wastewater Containing Congo Red Dye.

A rigorous knowledge of the bacterial growth kinetics is essential for the scaling-up and optimization of biodegradation process conditions in a bioreactor. Although a great deal of literature is available on the modeling of bacterial growth kinetics considering the inhibition at high substrate-loading, the inhibition caused by toxic metabolic byproducts was not accounted in the bacterial growth kinetics. This work primarily aimed at developing a parametric bacterial growth model to account for metabolite inhibition, indicated by a decelerating log-phase growth, which was rarely discussed in the previous studies. An efficient azo-dye degrading bacterium (Bacillus subtilis MN372379) was isolated from the sludge-waste nearby a carpet-dyeing unit. The isolated bacterial strain was used to decolorize the simulated wastewater containing Congo red dye. This study proposed a computational approach to calculate specific bacterial growth rate time-averaged over the entire sigmoidal log phase (including the decelerating phase) for incorporating the effect of metabolite-inhibition, in contrast to the conventional studies where only the initial part (accelerating) of log phase was considered. The nature of metabolite inhibition was also determined and found to be non-competitive. Next, the computed time-averaged specific bacterial growth rate was incorporated into three substrate inhibition models to account for both, the metabolite and substrate inhibitions, and subsequently their kinetic parameters were also determined. Finally, the initial dye concentration and inoculum size were optimized to yield maximum dye utilization rate. This study paves the way for predicting bacterial growth kinetic with improved accuracy to enable a better optimization of bioreactors at the industrial scale.

Purification of a potent mitogenic homodimeric Penicillium griseoroseum lectin and its characterisation.

Penicillium griseoroseum lectin was 80-fold purified by successive DEAE Sepharose anion exchange and Sephadex G-100 gel permeation chromatography. P. griseoroseum lectin exhibited haemagglutination activity towards protease-treated rabbit erythrocytes. It showed specificity towards various carbohydrates such as d-mannose, N-acetyl-d-glucosamine, mucins, and so forth. P. griseoroseum lectin was found as a glycoprotein with glycan content of 4.33%. Purified P. griseoroseum lectin is homodimeric having a molecular mass of 57 kDa with subunit molecular mass of 28.6 kDa. Haemagglutination activity of purified P. griseoroseum lectin was completely stable from 25°C to 35°C at a pH range of 6-7.5. Lectin activity was not influenced by divalent metal ions and denaturants. P. griseoroseum lectin manifested mitogenicity towards mice splenocytes and activity reached a peak at 75 µg/ml of lectin concentration. P. griseoroseum lectin in microgram concentrations stimulated proliferation of mice splenocytes. Thus, P. griseoroseum lectin exhibits potential mitogenicity, which can be exploited for further biomedical applications.

Temporal evolution of submicron particles during extreme fireworks.

Evolution of submicron particles in terms of particle number concentration and mobility-equivalent diameter was measured during Diwali festival-specific intensive pyrotechnic displays in Varanasi over central Indo-Gangetic Plain (IGP). A scanning mobility particle sizer coupled with an optical particle sizer was used to fit in an overlapping size range, and particle number concentration was analyzed to have an insight into the new particle formation and subsequent evolution of particles from nucleation to accumulation mode. Further, variation in black carbon (BC) concentration and aerosol ionic composition was measured simultaneously. Frequent fluctuation in particle number concentration in and around Diwali festival was evidenced, primarily influenced by local emission sources and meteorology, with three distinct peaks in number concentrations (dN/dlogDp, 3.1-4.5 × 104 cm3) coinciding well with peak firework emission period (18:00-23:00 h). Submicron particle size distribution revealed a single peak covering a size range of 80-130 nm, and for all instances, number concentration maximum coincided with geometric mean minimum, indicating the emission primarily in the ultrafine range (< 0.1 µm). Interestingly, during peak firework emissions, besides rise in accumulation mode, an event of new particle formation was identified with increase in nucleation and small Aitken mode, before being dispersed to background aerosols. On an integral scale, a clear distinction was noted between a normal and an episodic event, with a definite shift in the formation of ultrafine particles compared with the accumulation mode. The BC diurnal profile was typical, with a prominent nocturnal peak (12.0 ± 3.9 µg m-3) corresponding to a decrease in the boundary layer height. A slight variation in maximum BC concentration (16.8 µg m-3) was noted in the night of the event coinciding well with firework emissions. An increase in some specific ionic species was also noted in combination with an increase in the overall cation to anion ratio, which was explained in terms of heterogeneous transformation of NOx and catalytic conversion of SO2. Graphical abstract Time-resolved evolution of particle size distribution during normal and episodic events.

Thermostable and halotolerant keratinase from Bacillus aerius NSMk2 with remarkable dehairing and laundary applications.

Keratinases hydrolyze structural protein called keratin into constituent peptides. The present study reports excellent washing efficiency and dehairing properties of thermostable and halotolerant keratinase from Bacillus aerius NSMk2. Alkaline keratinase with molecular mass of 9 kDa displayed remarkable thermostability. K+ , Na+ , Ca2+ , Mn2+ , ß-mercaptoethanol, sodium sulfite, dithiothreitol, ethanol, isopropanol, Tween-20, and Tween-80 stimulated keratinase activity, while Hg2+ and Ba2+ were found to be inhibitory. The enzyme efficiently hydrolyzed a variety of complex protein substrates and exhibited high catalytic efficiency toward keratin-rich substrates and least toward collagen. Keratinase showed exceptional stability to salinity and was found to be compatible with most of the commercial detergents. Efficient removal of chocolate, blood, and egg albumin stains from clothes and tolerance to elevated temperature and salinity potentiated the suitability of keratinase from B. aerius NSMk2 as laundary additive. Keratinase could efficiently dehair goat skin after 15 hr of incubation without damaging the grain structure and collagen layers that assures its use as a promising contender for leather industry.

Response surface optimization of solid state fermentation for inulinase production from Penicillium oxalicum using corn bran.

Response surface methodology has been implemented for the utilization of corn bran for inulinase production by Penicillium oxalicum. CCRD of RSM with 15 runs was practiced to optimize three independent variables: moisture (70-90%), incubation time (4-8 days) and pH (5-8). However, other media constituents viz. inulin (1%), NaNO3 (0.2%), NH4H2PO4 (0.2%), KH2PO4 (0.2%), MgSO4·7H2O (0.05%) and FeSO4·7H2O (0.001%) were kept constant during solid state fermentations. Solid state fermentations were carried out at 30 °C at flask-level. A substantial inulinase production (77.95 IU/gds) was obtained under the optimized conditions i.e., moisture (80%), incubation time (6.0 days) and pH (6.5). Multiple correlation coefficient 'R2' for inulinase production was 1.00, which justifies good agreement between experimental and predicted values. Besides, 'R2' value close to one, also authenticates the validity of the model. The experimentation carried out at laboratory scale shown corn bran a good substrate for inulinase production by P. oxalicum.

Biocatalytic strategies in the production of galacto-oligosaccharides and its global status.

Galactooligosaccharides (GOSs) are the non-digestible carbohydrates that are composed of 3-10 or longer molecules of galactose and the terminal glucose molecule. These are considered as prebiotics owing to their various health benefits, and therefore is the major focus of research. These are generally synthesized by the catalytic activity of the glycoside hydrolases (GH) utilizing lactose as a substrate, which results in the mixture of GOSs differing in their degree of polymerization. Different microbial glycoside hydrolases have been used for the production. Moreover, to improve the production, different biotechnological strategies have been applied, such as use of immobilized enzymes or recombinant enzymes. Thus, this review discusses the different prospects of GOSs production and its purification techniques.

Efficacy of wood charcoal and its modified form as packing media for biofiltration of isoprene.

The efficacy of wood charcoal (WC) and nutrient-enriched wood charcoal (NWC) as biofilter packing media were assessed for isoprene biodegradation in a bioreactor comprising bioscrubber and a biofilter connected in series and inoculated with Pseudomonas sp. The bioreactors using WC and NWC exhibited >90% removal efficiency and around 369 g m-3 h-1 elimination capacity at around 404 g m-3 h-1 inlet loading rate. In both the bioreactors, the biofilter component showed better degradation capacity compared to the bioscrubber unit. The kinetic parameters, maximum elimination capacity, ECmax; substrate constant, Ks and ECmax/Ks for Michaelis-Menten model were evaluated. The lower Ks for the WC packed bioreactor indicated that ECmax achieved, was faster compared to others, while higher ECmax and ECmax/Ks for the NWC packed bioreactor suggests its superiority in isoprene abatement in the continuous mode. A comparison of the available published information on biofiltration of isoprene reflected polyurethane foam as the superior packing media.

Lectin activity in mycelial extracts of Fusarium species

ABSTRACT Lectins are non-immunogenic carbohydrate-recognizing proteins that bind to glycoproteins, glycolipids, or polysaccharides with high affinity and exhibit remarkable ability to agglutinate erythrocytes and other cells. In the present study, ten Fusarium species previously not explored for lectins were screened for the presence of lectin activity. Mycelial extracts of F. fujikuroi, F. beomiformii, F. begoniae, F. nisikadoi, F. anthophilum, F. incarnatum, and F. tabacinum manifested agglutination of rabbit erythrocytes. Neuraminidase treatment of rabbit erythrocytes increased lectin titers of F. nisikadoi and F. tabacinum extracts, whereas the protease treatment resulted in a significant decline in agglutination by most of the lectins. Results of hapten inhibition studies demonstrated unique carbohydrate specificity of Fusarium lectins toward O-acetyl sialic acids. Activity of the majority of Fusarium lectins exhibited binding affinity to D-ribose, L-fucose, D-glucose, L-arabinose, D-mannitol, D-galactosamine hydrochloride, D-galacturonic acid, N-acetyl-d-galactosamine, N-acetyl-neuraminic acid, 2-deoxy-D-ribose, fetuin, asialofetuin, and bovine submaxillary mucin. Melibiose and N-glycolyl neuraminic acid did not inhibit the activity of any of the Fusarium lectins. Mycelial extracts of F. begoniae, F. nisikadoi, F. anthophilum, and F. incarnatum interacted with most of the carbohydrates tested. F. fujikuroi and F. anthophilum extracts displayed strong interaction with starch. The expression of lectin activity as a function of culture age was investigated. Most species displayed lectin activity on the 7th day of cultivation, and it varied with progressing of culture age.

Lectin activity in mycelial extracts of Fusarium species.

Lectins are non-immunogenic carbohydrate-recognizing proteins that bind to glycoproteins, glycolipids, or polysaccharides with high affinity and exhibit remarkable ability to agglutinate erythrocytes and other cells. In the present study, ten Fusarium species previously not explored for lectins were screened for the presence of lectin activity. Mycelial extracts of F. fujikuroi, F. beomiformii, F. begoniae, F. nisikadoi, F. anthophilum, F. incarnatum, and F. tabacinum manifested agglutination of rabbit erythrocytes. Neuraminidase treatment of rabbit erythrocytes increased lectin titers of F. nisikadoi and F. tabacinum extracts, whereas the protease treatment resulted in a significant decline in agglutination by most of the lectins. Results of hapten inhibition studies demonstrated unique carbohydrate specificity of Fusarium lectins toward O-acetyl sialic acids. Activity of the majority of Fusarium lectins exhibited binding affinity to d-ribose, l-fucose, d-glucose, l-arabinose, d-mannitol, d-galactosamine hydrochloride, d-galacturonic acid, N-acetyl-d-galactosamine, N-acetyl-neuraminic acid, 2-deoxy-d-ribose, fetuin, asialofetuin, and bovine submaxillary mucin. Melibiose and N-glycolyl neuraminic acid did not inhibit the activity of any of the Fusarium lectins. Mycelial extracts of F. begoniae, F. nisikadoi, F. anthophilum, and F. incarnatum interacted with most of the carbohydrates tested. F. fujikuroi and F. anthophilum extracts displayed strong interaction with starch. The expression of lectin activity as a function of culture age was investigated. Most species displayed lectin activity on the 7th day of cultivation, and it varied with progressing of culture age.

Degradation kinetics and metabolites in continuous biodegradation of isoprene.

The kinetic parameters of isoprene biodegradation were studied in a bioreactor, comprising of bioscrubber and polyurethane foam packed biofilter in series and inoculated with Pseudomonas sp., using a Michaelis-Menten type model. The maximum elimination capacity, ECmax; substrate constant, Ks and ECmax/Ks values for bioscrubber were found to be 666.7 g m(-3) h(-1), 9.86 g m(-3) and 67.56 h(-1), respectively while those for biofilter were 3333 g m(-3) h(-1), 13.96 g m(-3) and 238.7 h(-1), respectively. The biofilter section exhibited better degradation efficiency compared to the bioscrubber unit. Around 62-75% of the feed isoprene got converted to carbon dioxide, indicating the efficient capability of bacteria to mineralize isoprene. The FTIR and GC-MS analyses of degradation products indicated oxidative cleavage of unsaturated bond of isoprene. These results were used for proposing a plausible degradation pathway for isoprene.

Sulfation of Aegle marmelos gum: synthesis, physico-chemical and functional characterization.

The present investigation was aimed at optimizing the conditions for preparing sulfated derivative of gum obtained from partially ripe fruits of Aegle marmelos. Elemental analysis, FTIR-ATR and NMR studies confirmed successful sulfation. The ratio of chlorosulfonic acid to pyridine exerted maximum influence on the degree of substitution followed by reaction temperature and reaction time. The sulfated derivative showed higher swelling in both acidic and alkaline pH as compared to unmodified gum. It also possessed higher negative zeta potential, higher viscosity, work of shear, firmness, consistency, cohesiveness and index of viscosity as compared to both unmodified gum as well as sodium alginate. Sulfated derivative was superior to unmodified gum and sodium alginate in terms of antimicrobial and anticoagulant activity. The sulfated sample appears to be a potential substitute over the unmodified gum sample and sodium alginate for modulating physicochemical properties of food and drug release dosage forms.

Community structure of methanogenic archaea and methane production associated with compost-treated tropical rice-field soil.

The diversity and density of methanogenic archaea and methane production were investigated ex situ at different growth stages of rice plant cultivated in compost-treated tropical rice fields. The qPCR analysis revealed variation in methanogens population from 3.40 × 10(6) to 1.11 × 10(7)  copies g(-1)  dws, in the year 2009 and 4.37 × 10(6) to 1.36 × 10(7)  copies g(-1)  dws in the year 2010. Apart from methanogens, a large number of bacterial (9.60 × 10(9) -1.44 × 10(10)  copies g(-1)  dws) and archaeal (7.13 × 10(7) -3.02 × 10(8)  copies g(-1)  dws) communities were also associated with methanogenesis. Methanogen population size varied in the order: flowering > ripening > tillering > postharvest > preplantation stage. The RFLP-based 16S rRNA gene-targeted phylogenetic analysis showed that clones were closely related to diverse group of methanogens comprising members of Methanomicrobiaceae, Methanosarcinaceae, Methanosaetaceae and RC I. Laboratory incubation studies revealed higher amount of cumulative CH(4) at the flowering stage. The integration of methanogenic community structure and CH(4) production potential of soil resulted in a better understanding of the dynamics of CH(4) production in organically treated rice-field soil. The hypothesis that the stages of plant development influence the methanogenic community structure leading to temporal variation in the CH(4) production has been successfully tested.

Pullulan: Microbial sources, production and applications.

Pullulan is a water-soluble glucan gum produced aerobically by growing a yeast like fungus Aureobasidium pullulans. It is a regularly repeating copolymer with the chemical structure {→6)-α-d-glucopyranosyl-(1→4)-α-d-glucopyranosyl-(1→4)-α-d-glucopyranosyl-(1→}n. Thus the polysaccharide is viewed as a succession of α-(1→6)-linked (1→4)-α-d-triglucosides i.e. maltotriose (G3). Pullulan have a wide range of commercial and industrial applications in many fields like food science, health care, pharmacy and even in lithography. Due to its strictly linear structure, pullulan is also very valuable in basic research as well as a well-defined model substance. This review attempts to critically appraise the current literature on fungal exopolysaccharide (EPS) 'pullulan' considering its microbial sources, structural geometry, upstream processing, downstream processing, peculiar characteristics and applications.