Phytochemical screening and antioxidative property evaluation of lipid-producing fungi.

The increased demands for natural bioactive compounds have induced the search for unusual sources. Microorganisms, especially fungi are a potent source of secondary metabolites, which could act mainly as antioxidant compounds to prevent oxidative stress. In the present study three soil-isolated fungi Aspergillus niger, Aspergillus heteromorphus and Aspergillus fumigatus, were screened for their oleaginous property as well as their potential for the production of bioactive compounds. Fungal biomasses were freeze dried and extracted with methanol using a cold percolation process for the production of intracellular metabolites and the fungal culture media after fermentation were examined for extracellular metabolites. Intracellular and extracellular extracts of the isolated fungi along with the single-cell oils extracted from those fungi were screened for phytochemicals, which showed the presence of alkaloids, flavonoides, glycosides, phenols, saponins and terpenoids. All strains showed potent antioxidant activity, determined using 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS) and ferric reducing antioxidant power (FRAP) activity. Extracellular extract and single-cell oil of A. heteromorphus showed the highest antioxidant activity with maximum ABTS radical scavenging activity and reducing potential. Highest content of phenolic and flavonoid compounds within the isolated fungi was found to be 37.58 mg gallic acid equivalent (GAE)/g and 62.07 mg catechine equivalent (CE)/g, respectively. Chromatographic analysis of the intracellular and extracellular extracts of the fungi showed the presence of gallic acid, di-hydroxy benzoic acid, ferulic acid, quercetin, epigerin, kampferol, trans cinnamic acid, chlorogenic acid and rutin, which made them biologically important and beneficial for human health.

Regio- and Diastereoselective [3 + 2]-Spiroannulation of Benzoxazines with Chalcones: A Rh(III)-Catalyzed Redox-Neutral Approach to α-Aroyl Spiro-Indanamines.

We report an atom-economic Rh(III)-catalyzed [3 + 2]-spiroannulation reaction between cyclic ketimines and α,ß-unsaturated carbonyl compounds, allowing the synthesis of novel spirocycles with concomitant generation of three stereogenic centers in one pot. The reaction does not require any silver additives or external oxidants and is believed to proceed in a redox-neutral manner. A broad substrate scope with good functional group tolerance permitted the synthesis of a vast spectrum of spirocyclic 1,4-benzoxazine derivatives containing polysubstituted α-aroyl-indanamines in good to excellent yields with high diastereoselectivity.

Optimization of the operation of packed bed bioreactor to improve the sulfate and metal removal from acid mine drainage.

The present study discusses the potentiality of using anaerobic Packed Bed Bioreactor (PBR) for the treatment of acid mine drainage (AMD). The multiple process parameters such as pH, hydraulic retention time (HRT), concentration of marine waste extract (MWE), total organic carbon (TOC) and sulfate were optimized together using Taguchi design. The order of influence of the parameters towards biological sulfate reduction was found to be pH > MWE > sulfate > HRT > TOC. At optimized conditions (pH - 7, 20% (v/v) MWE, 1500 mg/L sulfate, 48 h HRT and 2300 mg/L TOC), 98.3% and 95% sulfate at a rate of 769.7 mg/L/d. and 732.1 mg/L/d. was removed from the AMD collected from coal and metal mine, respectively. Efficiency of metal removal (Fe, Cu, Zn, Mg and Ni) was in the range of 94-98%. The levels of contaminants in the treated effluent were below the minimum permissible limits of industrial discharge as proposed by Bureau of Indian Standards (IS 2490:1981). The present study establishes the optimized conditions for PBR operation to completely remove sulfate and metal removal from AMD at high rate. The study also creates the future scope to develop an efficient treatment process for sulfate and metal-rich mine wastewater in a large scale.

Understanding the performance of sulfate reducing bacteria based packed bed reactor by growth kinetics study and microbial profiling.

A novel marine waste extract (MWE) as alternative nitrogen source was explored for the growth of sulfate reducing bacteria (SRB). Variation of sulfate and nitrogen (MWE) showed that SRB growth follows an uncompetitive inhibition model. The maximum specific growth rates (µmax) of 0.085 and 0.124 h(-1) and inhibition constants (Ki) of 56 and 4.6 g/L were observed under optimized sulfate and MWE concentrations, respectively. The kinetic data shows that MWE improves the microbial growth by 27%. The packed bed bioreactor (PBR) under optimized sulfate and MWE regime showed sulfate removal efficiency of 62-66% and metals removal efficiency of 66-75% on using mine wastewater. The microbial community analysis using DGGE showed dominance of SRB (87-89%). The study indicated the optimum dosing of sulfate and cheap organic nitrogen to promote the growth of SRB over other bacteria.

Reduction of start-up time through bioaugmentation process in microbial fuel cells using an isolate from dark fermentative spent media fed anode.

An electrochemically active bacteria Pseudomonas aeruginosa IIT BT SS1 was isolated from a dark fermentative spent media fed anode, and a bioaugmentation technique using the isolated strain was used to improve the start-up time of a microbial fuel cell (MFC). Higher volumetric current density and lower start-up time were observed with the augmented system MFC-PM (13.7 A/m(3)) when compared with mixed culture MFC-M (8.72 A/m(3)) during the initial phase. This enhanced performance in MFC-PM was possibly due to the improvement in electron transfer ability by the augmented strain. However, pure culture MFC-P showed maximum volumetric current density (17 A/m(3)) due to the inherent electrogenic properties of Pseudomonas sp. An electrochemical impedance spectroscopic (EIS) study, along with matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) analysis, supported the influence of isolated species in improving the MFC performance. The present study indicates that the bioaugmentation strategy using the isolated Pseudomonas sp. can be effectively utilized to decrease the start-up time of MFC.

Electrochemical C(sp3)-C(sp3) cross-dehydrogenative coupling: enabling access to 9-substituted fluorescent acridanes.

A straightforward strategy for direct access of C9-functionalized N-alkyl-acridanes in good to excellent yields has been established via a metal- and external oxidant-free sustainable electrochemical C(sp3)-C(sp3) cross-dehydrogenative coupling reaction between acridanes and benzo fused lactones at ambient temperature. A broad substrate scope with superior functional-group tolerance via anodic oxidation of acridanes permitted the synthesis of a vast spectrum of fluorescence-active acridanes with high quantum yields.

[3+2] Cascade Ring-Closing/Ring-Opening Strategy: Access to N-Indene-Tethered Amino Alcohols.

An unprecedented atom-economic redox neutral regioselective Rh(III)-catalyzed cascade [3+2] annulation of 2-aryl oxazoline with α,ß-unsaturated nitro olefins has been accomplished, furnishing a novel set of nitro-functionalized indene-tethered amino alcohols through a synergistic ring-closing/ring-opening strategy via the formation of two new C-C bonds and the regioselective cleavage of the C2-O bond of oxazoline under silver free mild reaction conditions with a broad substrate scope.

Synthesis of indene-fused spiro-dibenz(ox)azepines via Rh(III)-catalyzed cascade regioselective C-H activation/annulation.

We report an unprecedented atom-economic one-pot Cp*Rh(III)-catalyzed regioselective [3+2]-spiroannulation reaction between dibenz(ox)azepines and ynones, allowing the synthesis of biologically relevant novel spirocyclic dibenz(ox)azepines under operationally simple and mild reaction conditions. The reaction proceeds without any silver additive or external oxidant implementing a redox-neutral pathway. A broad substrate scope with diverse functional group tolerance permitted the regioselective synthesis of a wide spectrum of indene-containing spirocyclic dibenz(ox)azepines in good to excellent yields. Also, we showcased detailed mechanistic studies to justify the formation of spirocycles. In addition, the synthetic utility of this process was also demonstrated by the modular synthesis of various steroid conjugates.

A condition-tuned unorthodox approach to indole-3-carboxylic acids and anthranilic acids via carbon atom translocation.

We describe a robust one-pot cascade method for the synthesis of indole-3-carboxylic acids using isatins and DMSO via a one-carbon translocation involving in situ generation of α,ß-unsaturated methylvinylsulfoxide followed by amide bond cleavage and ring closure. The methodology has been extended to afford anthranilic acid derivatives by tuning the reaction conditions in the presence of molecular oxygen. Importantly, easy access to commercially available drugs, including tropisetron, is demonstrated.

A radical-radical cross-coupling reaction of xanthene with sulfonyl hydrazides: facile access to xanthen-9-sulfone derivatives.

A straightforward strategy for direct incorporation of sulfonyl units into a xanthene moiety for accessing xanthen-9-sulfone derivatives in good to excellent yields has been established via metal-free radical-radical cross-coupling reaction of xanthenes and sulfonyl hydrazides. Using easily accessible starting materials, this methodology proceeds efficiently with a high degree of functional group compatibility and with a wide scope of both xanthenes and sulfonyl hydrazides under operationally simple reaction conditions. Mechanistic investigations revealed that sulfonyl radicals could be generated from sulfonyl hydrazides in the presence of TBHP under an oxygen atmosphere.

Effects of in utero arsenic exposure on child immunity and morbidity in rural Bangladesh.

Chronic exposure to arsenic, a potent carcinogen and toxicant, via drinking water is a worldwide public health problem. Because little is known about early-life effects of arsenic on immunity, we evaluated the impact of in utero exposure on infant immune parameters and morbidity in a pilot study. Pregnant women were enrolled at 6-10 weeks of gestation in Matlab, a rural area of Bangladesh, extensively affected by arsenic contamination of tubewell water. Women (n=140) delivering at local clinics were included in the study. Anthropometry and morbidity data of the pregnant women and their children, as well as infant thymic size by sonography were collected. Maternal urine and breast milk were collected for immune marker and arsenic assessment. Maternal urinary arsenic during pregnancy showed significant negative correlation with interleukin-7 (IL-7) and lactoferrin (Ltf) in breast milk and child thymic index (TI). Urinary arsenic was also positively associated with fever and diarrhea during pregnancy and acute respiratory infections (ARI) in the infants. The effect of arsenic exposure on ARI was only evident in male children. The findings suggest that in utero arsenic exposure impaired child thymic development and enhanced morbidity, probably via immunosuppression. The effect seemed to be partially gender dependent. Arsenic exposure also affected breast milk content of trophic factors and maternal morbidity.

Novel integration of biohydrogen production with fungal biodiesel production process.

An integration of bio-H2 with fungal biodiesel production process was investigated. Highest cumulative H2 production of 3.3 ±â€¯0.20 L L-1 was observed during media optimization using mixture design. Using optimized media composition, continuous H2 production at 0.2 h-1 dilution rate, showed highest H2 production rate, H2 yield and biomass yield of 1020 ±â€¯23 mL L-1 h-1, 2.8 ±â€¯0.1 mols mol-1 reducing sugar and 1.2 ±â€¯0.06 g L-1, respectively. Using the spent media generated from the dark fermentation, oleaginous yeast cultivation was done. Highest biomass and total lipid yield of 6.4 ±â€¯0.20 g L-1 and 0.46 ±â€¯0.04 g g-1 was observed at initial 15% v/v inoculums strength, pH of 5, 1.5 L min-1 aeration rate and 25 °C temperature of cultivation, respectively. Energy recovery improved by 90.3% in integrated process when compared with single stage hydrogen production.

Enhancement in lipid content of Chlorella sp. MJ 11/11 from the spent medium of thermophilic biohydrogen production process.

The present study investigates the effect of spent media of acetogenic dark fermentation for mixotrophic algal cultivation for biodiesel production. Mixotrophic growth conditions were optimized in culture flask (250mL) using Chlorella sp. MJ 11/11. Maximum lipid accumulation (58% w/w) was observed under light intensity, pH, nitrate and phosphate concentration of 100µmolm-2s-1, 7, 2.7mM and 1.8mM, respectively. Air lift (1.4L) and flat panel (1.4L) reactors were considered for algal cultivation. Air lift showed significant improvement in biomass and lipid production as compared to flat panel reactor. The results could help in development of sustainable technology involving acetogenic hydrogen production integrated with sequential mitigation of spent media by algal cultivation for improved energy recovery.

Biohythane production from organic wastes: present state of art.

The economy of an industrialized country is greatly dependent on fossil fuels. However, these nonrenewable sources of energy are nearing the brink of extinction. Moreover, the reliance on these fuels has led to increased levels of pollution which have caused serious adverse impacts on the environment. Hydrogen has emerged as a promising alternative since it does not produce CO2 during combustion and also has the highest calorific value. The biohythane process comprises of biohydrogen production followed by biomethanation. Biological H2 production has an edge over its chemical counterpart mainly because it is environmentally benign. Maximization of gaseous energy recovery could be achieved by integrating dark fermentative hydrogen production followed by biomethanation. Intensive research work has already been carried out on the advancement of biohydrogen production processes, such as the development of suitable microbial consortium (mesophiles or thermophiles), genetically modified microorganism, improvement of the reactor designs, use of different solid matrices for the immobilization of whole cells, and development of two-stage process for higher rate of H2 production. Scale-up studies of the dark fermentation process was successfully carried out in 20- and 800-L reactors. However, the total gaseous energy recovery for two stage process was found to be 53.6 %. From single-stage H2 production, gaseous energy recovery was only 28 %. Thus, two-stage systems not only help in improving gaseous energy recovery but also can make biohythane (mixture of H2 and CH4) concept commercially feasible.

Electro-stimulated microbial factory for value added product synthesis.

Interplay of charge between bacteria and electrode has led to emergence of bioelectrochemical systems which leads to applications such as production of electricity, wastewater treatment, bioremediation and production of value added products. Many electroactive bacteria have been identified that have unique external electron transport systems. Coupling of electron transport with carbon metabolism has opened a new approach of carbon dioxide sequestration. The electron transport mechanism involves various cellular and sub cellular molecules. The outer membrane cytochromes, Mtr-complex and Ech-complex are few key molecules involved in electron transport in many electrogenic bacteria. Few cytochrome independent acetogenic electroactive bacteria were also discovered using Rnf complex to transport electrons. For improved productivity, an efficient bioreactor design is mandatory. It should encompass all certain critical issues such as microbial cell retention, charge dissipation, separators and simultaneous product recovery.

Comparative evaluation of the hydrogen production by mixed consortium, synthetic co-culture and pure culture using distillery effluent.

Wastewater comprises of various carbon sources. So, the use of microbial consortium may improve the hydrogen production and organic reduction. The present study deals with biohydrogen production by acidogenic mixed consortia (AMC), synthetic co-culture (Klebsiella pneumoniae IIT-BT 08 and Citrobacter freundii IIT-BT L139) and pure culture using distillery effluent (DE). Higher hydrogen yield was observed in case of AMC (9.17 mol/kg CODreduced) as compared to the synthetic co-culture and pure culture. PCR-DGGE analysis indicated that the consortium was predominated by species closely affiliated to Clostridium sp. The average hydrogen production rate was 267 mL/Lh. The maximum hydrogen production rate (Rm), hydrogen production potential (P) and lag time (λ) by AMC using DE were 507.2 mL/Lh, 3729 m/L and 2.04 h, respectively. Maximum gaseous energy recovery by AMC was found to be higher by 21.9% and 45.4% than that of using co-culture and pure culture respectively.

Improvement of biomass production by Chlorella sp. MJ 11/11 for use as a feedstock for biodiesel.

Algal biomass is gaining importance for biofuel production as it is rich in lipids. It becomes more significant when biomass is produced by capturing atmospheric greenhouse gas, CO2. In the present study, the effect of different physicochemical parameters were studied on the biomass and lipid productivity in Chlorella sp. MJ 11/11. The different parameters viz. initial pH, nitrate concentration, and phosphate concentration were optimized using single-parameter studies. The interactions between the parameters were determined statistically using the Box-Behnken design of optimization. The optimal values were decided by analyzing them with response surface methodology. The optimum levels of the parameters (pH 6.5, nitrate concentration 0.375 g L(-1), and phosphate concentration 0.375 mL L(-1)) yielded a maximum biomass concentration of 1.26 g L(-1) at a constant light intensity of 100 µmol m(-2) s(-1) and temperature of 30 °C. The effect of CO2 concentration on the biomass production was also investigated and was found to be a maximum of 4 g L(-1) at 5 % air-CO2 mixture (v/v). Maximum lipid content of 24.6 % (w/w) was observed at 2 % air-CO2 mixture (v/v). Fatty acid analyses of the obtained algal biomass suggested that they could be a suitable feedstock for biodiesel production.

Improvement of the degradation of sulfate rich wastewater using sweetmeat waste (SMW) as nutrient supplement.

External dosing of sweetmeat waste (SMW) dosing into exhausted upflow packed bed bioreactor (PBR) resulted in prompt reactivation of SO4(2-) removal. Different SMW concentrations in terms of chemical oxygen demand (COD)/SO4(2-) ratios (1, 2, 4 and 8) were introduced into four identical PBR where process stability was found within 3 weeks of operation. SO4(2-) removal was proportional to COD/SO4(2-) ratios up to 4 at which maximum sulfate removal (99%) was achieved at a rate of 607 mg/d. The value of COD consumption:SO4(2-)removal was much higher at ratio 4 than 8 whereas, ratio 2 was preferred over all. Net effluent acetate concentration profile and total microbial population attached to the reactor matrices were corresponding to COD/SO4(2-) ratio as 4>8>2>>1. Sulfate reducing bacteria (SRB) population was found to be inversely proportional to COD/SO4(2-) ratio in which acetate oxidizing SRB and fermentative bacteria were the dominant.

Improvement of power generation using Shewanella putrefaciens mediated bioanode in a single chambered microbial fuel cell: effect of different anodic operating conditions.

Three different approaches were employed to improve single chambered microbial fuel cell (sMFC) performance using Shewanella putrefaciens as biocatalyst. Taguchi design was used to identify the key process parameter (anolyte concentration, CaCl2 and initial anolyte pH) for maximization of volumetric power. Supplementation of CaCl2 was found most significant and maximum power density of 4.92 W/m(3) was achieved. In subsequent approaches, effect on power output by riboflavin supplementation to anolyte and anode surface modification using nano-hematite (Fe2O3) was observed. Volumetric power density was increased by 44% with addition of 100 nM riboflavin to anolyte while with 0.8 mg/cm(2) nano-Fe2O3 impregnated anode power density and columbic efficiency increased by 40% and 33% respectively. Cyclic voltammetry revealed improvement in electrochemical activity of Shewanella with nano-Fe2O3 loading and electrochemical impedance depicted inverse relationship between charge transfer resistance and nano-Fe2O3 loading. This study suggests anodic improvement strategies for maximization of power output.

Continuous mode of carbon dioxide sequestration by C. sorokiniana and subsequent use of its biomass for hydrogen production by E. cloacae IIT-BT 08.

The present study investigated to find out the suitability of the CO2 sequestered algal biomass of Chlorella sorokiniana as substrate for the hydrogen production by Enterobacter cloacae IIT-BT 08. The maximum biomass productivity in continuous mode of operation in autotrophic condition was enhanced from 0.05 g L(-1) h(-1) in air to 0.11 g L(-1) h(-1) in 5% air-CO2 (v/v) gas mixture at an optimum dilution rate of 0.05 h(-1). Decrease in steady state biomass and productivity was less sensitive at higher dilution and found fitting with the model proposed by Eppley and Dyer (1965). Pretreated algal biomass of 10 g L(-1) with 2% (v/v) HCl-heat was found most suitable for hydrogen production yielding 9±2 mol H2 (kg COD reduced)(-1) and was found fitting with modified Gompertz equation. Further, hydrogen energy recovery in dark fermentation was significantly enhanced compared to earlier report of hydrogen production by biophotolysis of algae.