Various lactic acid bacteria (LAB) have been isolated and screened for biosurfactant production and their biomedical and food applications. Additionally, various different concentrations of the biosurfactant (0.625-25 mg ml-1) were used to evaluate its antimicrobial and antiadhesive potential against a range of pathogenic microorganisms. Biosurfactant was found to be stable to pH changes over a range of 4.0-12.0, being most effective at pH 7 and showed no apparent loss of surface tension and emulsification efficiency after heat treatment at 125 °C for 15 min. Present study demonstrated that biosurfactant obtained from Lactobacillus helveticus has the ability to counteract effectively the initial deposition of biofilm forming pathogens to silicone surfaces and to significantly sluggish biofilm growth.

Epigenetics have witnessed a renewed interest over the past decade and assays with recombinant histones has become an important tool for uncovering various aspects of histone biology. However, at times absence of recombinant histone accumulation in bacteria is encountered which is also commonly observed for many eukaryotic proteins in general. In this study, we have investigated the effect of multiple parameters on heterologous expression of proteins. We show that there is marked variability in the accumulation of H2A.2, H2B.1, H3.2 and H4 in the recombinant host, possibly owing to translational variability and degradation by the host proteases. We found that the variability could be overcome by incorporation of the commonly used purification tags, like GST or MBP, of appropriate size and position. Our results provide compelling evidence that transcript parameters like rare codon and GC content, mRNA secondary structure etc. together modulate translation kinetics and govern recombinant protein accumulation.

A novel thermophilic bacterial strain of the genus Aeribacillus was isolated from Thar Dessert Pakistan. This strain showed significant antibacterial activity against Micrococcus luteus, Staphylococcus aureus, and Pseudomonasaeruginosa. The strain coded as 'SAT4' resembled with Aeribacillus pallidus in the morphological, biochemical and molecular tests. The production of antibacterial metabolites by SAT4 was optimized. These active metabolites were precipitated by 50% ammonium sulphate and purified through sephadex G-75 gel permeation chromatography and reverse phase HPLC. The molecular weight of 37 kDa was examined by SDS-PAGE. The structural elucidation of the purified product was studied by FTIR, 1H and 13C NMR. The X-ray diffractions study showed that the crystals belonged to the primitive orthorhombic lattice (a = 12.137, b = 13.421, c = 14.097 Å) and 3D structure (proposed name: Aeritracin) was determined. This new peptide antibacterial molecule can get a position in pharmaceutical and biotechnological industrial research.

Leishmaniasis, a disease of the developing world affects about 12 million people and has limited therapeutic interventions available. L-type lectins, Endoplasmic Reticulum Golgi Intermediate Compartment/Vesicular Integral Proteins (ERGIC-53/VIP36) are involved in protein sorting in luminal compartments of animal cells and are important for parasite biology. A lectin homologue was identified through a bioinformatics analysis of Leishmania genome and it was found to have N-terminal conserved carbohydrate recognition domain (CRD) and a unique C-terminal region rich in repetitive amino acids and a poly glutamine tract. The N-terminal CRD region was cloned and expressed in Escherichia coli, but gave an insoluble expression which was re-solubilized by on column refolding. The fold integrity was checked through CD, fluorescence and functional assay of hemagglutination activity using rabbit erythrocyte. Bioinformatics analysis identified 15 members from Tritryps (Leishmania spp., Trypanosoma spp.) and they separate out as a distinct clade in the global phylogenetic analysis of all ERGIC-53/VIP36 sequences downloaded from Uniprot. Our analysis shows that the extended C-terminal regions with repeats is unique to Tritryps and this repeat pattern is different in sequences from Leishmania spp. and Trypanosoma spp. and all these features make this protein an interesting candidate for further detailed studies.

A single step protocol to produce biofunctionalized silver nanoparticles (AgNPs) using the aqueous extract of Cassia fistula flower as "natural factory" was investigated. The reaction between silver ions and aqueous flower extract after the bioreduction process has resulted in the formation of reddish brown color colloidal solution. XRD pattern showed the face centered cubic crystalline structure of AgNPs and exhibited spherical morphology as characterized by FE-SEM. FTIR studies identified different functional groups involved in effective capping of AgNPs. The zeta potential affirmed the phytoreduced AgNPs possess good stability and the size of the particle was measured by DLS. The synthesized AgNPs displayed effective cytotoxic potential against MCF7 and the inhibitory concentration (IC50) was recorded at 7.19 μg/mL. The apoptotic effects of the AgNPs were also confirmed by AO/EB staining. The investigation presents preliminary evidence that biosynthesized AgNPs can be used in the development of novel anticancer drugs.

Novel enzyme-luminescence method is used for the rapid and sensitive in vitro detection of natural neurotoxins (e.g., shellfish and mushroom toxins) using model brain cells. Paralytic shellfish poisons gonyautoxins (e.g., GTX2,3 and GTX1,4) were detected at 1 nM level by their inhibition of glutamate release from C6 glioma cells upon drug stimulation (IC50: GTX2,3 = 30 nM and GTX1,4 = 8 nM). Activation of glutamate release from C6 cells by ibotenic acid (a mushroom toxin) was also evaluated (EC50 = 10 nM). The method was tested for real-time detection of glutamate release from primary rat cortical neurons. Dose-dependent effects of KCl (0-200 mM) and NMDA on glutamate release from primary cortical neurons were studied. The effects of different culture conditions on K+-depolarization-induced glutamate release were also investigated. The method may be applicable to screening of drugs and toxins, and finding glutamatergic neurons in brain slices without in situ staining.

The study was conducted to evaluate the conditions to enhance the accumulation of lipids and starch in Chlorococcum sp. TISTR 8583 for the production of biofuel. The Chlorococcum sp. TISTR 8583 was cultivated on BG-11 medium under optimized light intensity. The nitrogen limitation (NL) enhanced the accumulation of both starch and lipids and resulted in 34.02% total sugars as compared to 22.57% on nitrogen supplemented (NS) media only. Similarly, the nitrogen supplemented (NS) media produced 17.05% lipids as compared to 29.59% lipids by NL media. The biomass was investigated for biodiesel and bioethanol production by adopting different pretreatment strategies, such as enzyme, acid and alkaline pretreatments. The alkaline pretreatment was found to be efficient strategy (23.67 wt% sugars/g algal biomass: 1.2% (w/v) at 140 ⁰C for 30 min) while the acid pretreatment (1%: v/v; 140 °C) was least effective pretreatment strategy with the yield of 14.83 wt% sugars/g algal biomass.

The β-1,4-d-xylanohydrolase is an industry valuable catalytic protein and used to synthesize xylooligosaccharides and xylose. In the current study, β-1,4-d-xylanohydrolase from Geobacillus stearothermophilus KIBGE-IB29 was partially purified up to 9.5-fold with a recovery yield of 52%. It exhibited optimal catalytic activity at pH-7.0 and 50 °C within 5 min. Almost 50% activity retained at pH-4.0 to 9.0 however, 70% activity observed within the range of 40 °C to 70 °C. The β-1,4-d-xylanohydrolase showed a significant hydrolytic pattern with 48.7 kDa molecular mass. It was found that the enzymatic activity improved up to 160% with 1.0 mM ethanol. Moreover, the activity of enzyme drastically increased up to 2.3 and 1.5 fold when incubated with Tween 80 and Triton X-100 (1.0 mM), respectively. The β-1,4-d-xylanohydrolase also retained 72% activity at -80 °C after 180 days. Such a remarkable biochemical properties of β-1,4-d-xylanohydrolase make it possible to forecast its potential use in textile and food industries.

Dairy processing generates large volumes of wastewater that require extensive nutrient remediation prior to discharge. Significant commercial opportunities exist therefore for cost-effective biotechnologies capable of achieving this requirement. In this study the authors evaluated the use of intermittently aerated sequencing batch reactors, (IASBRs), as a single-tank biotreatment system for co-removal of COD, nitrogen and phosphorus from synthetic dairy processing wastewater. Variation of the IASBR aeration rates, (0.8, 0.6 and 0.4 L/min), had significant impacts on the respective nutrient removal efficiencies and underlying microbial diversity profiles. Aeration at 0.6 L/min was most effective and resulted in >90% co-removal of orthophosphate and ammonium. 16S rRNA based pyrosequencing of biomass DNA samples revealed the family Comamonadaceae was notably enriched (>80% relative abundance) under these conditions. In silico predictive metabolic modelling also identified Comamonadaceae as the major contributor of several known genes for nitrogen and phosphorus assimilation (nirK, nosZ, norB, ppK, ppX and phbC).

Echinocandin B is a potent antifungal against the majority of fungal pathogens and its biosynthesis occurred by ecd and hty gene clusters in Emericella rugulosa NRRL 11440. We elucidated the functional necessity of in-clustered transcription factor; ecdB in the production of echinocandin B. We deleted the ecdB gene and found that ΔecdB mutant has no significant effect on echinocandin B production. The expression level of most of the ecd and hty cluster genes was not significantly altered except few of them up-regulated in knockout strain. The complete abrogation in ecdB gene expression was observed in ΔecdB strain. However, the interactions of purified EcdB protein with DNA sequence of ecdA, ecdH, ecdK and ecdI promoter was confirmed in-vitro. Our results conclude that EcdB protein in-vitro binds to the ecdA, ecdH, ecdK and ecdI promoter but in-vivo, it could not significantly affect the gene expression and echinocandin B production in Emericella rugulosa.

Characterization, antioxidant and immunomodulatory potential of exopolysaccharide (EPS) produced by wild type and mutant Weissella confusa was investigated. The EPS production ranged from 5490.2 to 5580.7 mg/L. Wild type Weissella confusa (WWCEPS) had the highest EPS production. Eight (8) sugar moieties were present in the EPS. Galactose had the highest EPS composition (34.6 mg/100 g and 33.5 mg/100 g EPS) in Wild type Weissella confusa EPS (WWCEPS) and mutant Weissella confusa EPS (MWCEPS). Wild type Weissella confusa and mutant Weissella confusa EPS had antioxidant capacity. The scavenging assay for the antioxidant increased in a dose dependent (0.5-10 mg/mL) manner. Wild type Weissella confusa EPS had the highest 1,1-Diphemy 1-2-picryl-hydrazyl (DPPH) capacity, total antioxidant activity, hydrogen peroxide and reducing power activity (71%, 1.9%, 86.7% and 1.9%). The mice injected peritorially with mutant Weissella confusa EPS had the highest IgG and IgM (68-87 mg/dL and 64-70 mg/dL). IgA of the mice treated with Wild type Weissella confusa EPS increased from 67 to 73 mg/dL. Wild type and mutant Weissella confusa EPS had immunomodulatory activity on the treated mice.

The rationale of the present study was to isolate and identify endophytic fungi from endemic medicinal plants in Eastern Ghats and screened for antimicrobial potential of isolated fungal crude extracts. A total of 329 endophytic strains were isolated from 600 infected leaves and stem cuttings of endemic plants. The diversity and species richness was analyzed statistically and found to be higher in leaf segments than in stem segments. From isolated fungal strains, Cladosporium delicatulum was identified using molecular identification methods and selected as the most potent plumbagin-producing endophytic strain. Further the isolation and structural characterization of endophytic fungal plumbagin (5-hydroxyl-2-methylnaptalene-1,4-dione) was purified and confirmed through spectroscopy analysis. The molecular weight was determined as m/z 188 in positive mode by ESI-MS, which confirmed to be plumbagin which potentially inhibited all tested pathogens, therefore the endophytic fungal plumbagin from the current study possesses important biological activities against pathogens.

Phenylalanine ammonia-lyase (RgPAL) from Rhodotorula glutinis JN-1 stereoselectively catalyzes the conversion of the l-phenylalanine into trans-cinnamic acid and ammonia, and was used in chiral resolution of dl-phenylalanine to produce the d-phenylalanine under acidic condition. However, the optimum pH of RgPAL is 9 and the RgPAL exhibits low catalytic efficiency at acidic side. Therefore, a mutant RgPAL with a lower optimum pH is expected. Based on catalytic mechanism and structure analysis, we constructed a mutant RgPAL-Q137E by site-directed mutagenesis, and found that this mutant had an extended optimum pH 7-9 with activity of 1.8-fold higher than that of the wild type at pH 7. As revealed by Friedel-Crafts-type mechanism of RgPAL, the improvement of the RgPAL-Q137E might be due to the negative charge of Glu137 which could stabilize the intermediate transition states through electrostatic interaction. The RgPAL-Q137E mutant was used to resolve the racemic dl-phenylalanine, and the conversion rate and the eeD value of d-phenylalanine using RgPAL-Q137E at pH 7 were increased by 29% and 48%, and achieved 93% and 86%, respectively. This work provides an effective strategy to shift the optimum pH which is favorable to further applications of RgPAL.

The present study emphasizes the biosurfactant mediated anthracene degradation by a marine alkaliphile Bacillus licheniformis (MTCC 5514). The isolate, MTCC 5514 degraded >95% of 300 ppm anthracene in an aqueous medium within 22 days and the degradation percentage reduced significantly when the concentration of anthracene increased to above 500 ppm. Naphthalene, naphthalene 2-methyl, phthalic acid and benzene acetic acid are the products of degradation identified based on thin layer chromatography, high performance liquid chromatography, gas chromatography and mass analyses. It has been observed that the degradation is initiated by the biosurfactant of the isolate for solubilization through micellation and then the alkali pH and intra/extra cellular degradative enzymes accomplish the degradation process. Encoding of genes responsible for biosurfactant production (licA3) as well as catabolic reactions (C23O) made with suitable primers designed. The study concludes in situ production of biosurfactant mediates the degradation of anthracene by B. licheniformis.

Cyclic-diGMP is a bacterial messenger that regulates many physiological processes, including many attributed to pathogenicity. Bacteria synthesize cyclic-diGMP from GTP using diguanylate cyclases; its hydrolysis is catalyzed by phosphodiesterases. Here we report the over-expression and purification of a bi-functional diguanylate cyclase-phosphodiesterase from Agrobacterium vitis S4. Using homology modeling and primary structure alignment, we identify several amino acids predicted to participate in the phosphodiesterase reaction. Upon altering selected residues, we obtain variants of the enzyme that efficiently and quantitatively catalyze the synthesis of cyclic-diGMP from GTP without hydrolysis to pGpG. Additionally, we identify a variant that produces cyclic-diGMP while immobilized to NiNTA beads and can catalyze the conversion of [α-32P]-GTP to [32P]-cyclic-diGMP. In short, we characterize a novel cyclic-diGMP processing enzyme and demonstrate its utility for efficient and cost-effective production of cyclic-diGMP, as well as modified cyclic-diGMP molecules, for use as probes in studying the many important biological processes mediated by cyclic-diGMP.

Among four crude oil-degrading fungi strains that were isolated from a petroleum-polluted area in the Rumaila oil field, two fungi strains showed high activity in aliphatic hydrocarbon degradation. ITS sequencing and analysis of morphological and biochemical characteristics identified these strains as Penicillium sp. RMA1 and RMA2. Gravimetric and gas chromatography analysis of the crude oil remaining in the culture medium after 14 days of incubation at 30 °C showed that RMA1 and RMA2 degraded the crude oil by 57% and 55%, respectively. These strains reduced surface tension when cultured on crude oil (1% v/v) and exhibited a cell surface hydrophobicity of more than 70%. These results suggested that RMA1 and RMA2 performed effective crude oil-degrading activity and crude oil emulsification. In conclusion, these fungal strains can be used in bioremediation process and oil pollution reduction in aquatic ecosystems.

Triphala, an Indian ayurvedic triherbal formulation, is an equiproportional mixture of fruits of three herbs, amalaki (Emblica officinalis), haritaki (Terminalia chebula) and bibhitaki (Terminalia bellerica). The present study focused on phytocompounds detection and comparative analysis of various biochemical activities in the aqueous and methanolic extracts of triphala and its constituting herbs. Antioxidant activity was determined by 1, 1-diphenyl-2-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), super oxide dismutase (SOD), catalase assay. Antibacterial potential was determined by broth dilution and agar well diffusion assays. Results revealed the presence of valuable bioactive compounds such as flavonoids, alkaloids, phenols, etc which might be responsible for biochemical activities. Extracts exhibited satisfactory radical-scavenging activity comparable with ascorbic acid. Methanolic extracts demonstrated higher antioxidant activity compared to aqueous extract. Extracts showed promising antibacterial potential against tested strain comparable to ampicillin. Hence, it can be concluded that triphala may be a promising candidate in pharmaceuticals and future medicine.

Proteiniphilum saccharofermentans str. M3/6T is a recently described species within the family Porphyromonadaceae (phylum Bacteroidetes), which was isolated from a mesophilic laboratory-scale biogas reactor. The genome of the strain was completely sequenced and manually annotated to reconstruct its metabolic potential regarding biomass degradation and fermentation pathways. The P. saccharofermentans str. M3/6T genome consists of a 4,414,963 bp chromosome featuring an average GC-content of 43.63%. Genome analyses revealed that the strain possesses 3396 protein-coding sequences. Among them are 158 genes assigned to the carbohydrate-active-enzyme families as defined by the CAZy database, including 116 genes encoding glycosyl hydrolases (GHs) involved in pectin, arabinogalactan, hemicellulose (arabinan, xylan, mannan, β-glucans), starch, fructan and chitin degradation. The strain also features several transporter genes, some of which are located in polysaccharide utilization loci (PUL). PUL gene products are involved in glycan binding, transport and utilization at the cell surface. In the genome of strain M3/6T, 64 PUL are present and most of them in association with genes encoding carbohydrate-active enzymes. Accordingly, the strain was predicted to metabolize several sugars yielding carbon dioxide, hydrogen, acetate, formate, propionate and isovalerate as end-products of the fermentation process. Moreover, P. saccharofermentans str. M3/6T encodes extracellular and intracellular proteases and transporters predicted to be involved in protein and oligopeptide degradation. Comparative analyses between P. saccharofermentans str. M3/6T and its closest described relative P. acetatigenes str. DSM 18083T indicate that both strains share a similar metabolism regarding decomposition of complex carbohydrates and fermentation of sugars.

The biosurfactant production is characterized by high costs with substrates, which does not make them sufficiently competitive against synthetic surfactants. The insertion of alternative sources of low cost, especially agro-industrial residue, is an excellent alternative to make this competitiveness viable. An alkaline pretreatment was used to extract the hemicellulose from corncob in order to enhance its C5 fraction, common to vegetable biomasses. The hemicellulosic corncob liquor was used with glucose and mineral salt solution as carbon and nutrients sources in a fermentation process for the growth of Bacillus subtilis. It was performed a 23 full factorial design to determine the best conditions for the surfactin production in relation to the following response variables: surface tension reduction rate (STRR) and emulsification index (EI24), from which were obtained two optimized bioproducts under specific conditions. The optimized biosurfactants found to be effected presenting a critical micelle concentration of 100 mg.L-1 and a maximum bioremediation potential of 85.18%, as well as maximum values of 57.38% and 65.30% for STRR and EI24 variables, respectively. Overall results pointed for a successful commercial application for the surfactin produced.

Given the upwelling of a variety of potential applications laccases could participate in, it would be fitting to equally make available laccases that are well suited for the aforementioned. Therefore historian understanding of the catalytic and physicochemical properties is desirable. Owing to this, the biochemical properties of the crude laccases from Achromobacter xylosoxidans HWN16 (Hb9c) and Citrobacter freundii LLJ 16 (Ie1c) were assessed. Furthermore, a hint of the molecular basis for their production from respective organisms was presented. Results showed that both laccases were tolerant, and sometimes had their activities improved by the set of parameters tested. They were active at broad range of temperature (0-90 °C), pH (3-11), and were equally thermo- and pH-stable. Their activities were either improved, or left unabated by cations, detergents, and chloride (5-40%), however, the highlight of the study was their augmented activity, when they were incubated with certain concentrations of fluoride (2-20%), a potent inhibitor. They were depicted to have multiple homologous laccase encoding genes, on molecular evaluation, which may be responsible the conferral of these remarkable qualities they possess. Therefore, the laccases might be beneficial, if employed in formulations for a wide range of environmental and biotechnological applications. Moreover, the molecular machinery of their production be exploited for economical benefits in the immediate future.