Screening and optimization of arsenic degrading bacteria and their potential role in heavy metal bioremediation

Industrialization has added extremely toxic metalloid arsenic into the environment which at high concentration severely threatens the biota. Naturally, some microbes possess the ability to bio-accumulate metals and also to transform arsenite (As III) a toxic form to a non-toxic arsenate As V. The present study aimed to isolate arsenic resistant bacterias from the arsenic contaminated soil and water. Among eleven bacterial isolates, three FAs 1, 4 and 9 exhibited tolerance against sodium arsenite at 100mM concentration by achieving growth of 7.48×109,1.57×109 and 2.23×109 C.F.U./ml, respectively. Optimization at different conditions such as temperature, pH and arsenic concentration revealed high arsenic tolerance from isolate FAs 4 (5.33×108) at 37°C and FAs 1 (4.43×108 C.F.U./ml) at pH 7. Arsenic resistance at optimum conditions for the bacterial strains FAs 1, FAs 4 and FAs 9 showed maximum growth at 80mM concentration of arsenite. These bacterial isolates did not show redox ability to oxidize arsenite As III to arsenate As V. However bacterial isolates FAs 1, FAs 4 and FAs 9 were able to accumulate arsenic 39.16, 148 and 125 µg/L on the 4th, 3rd and 5th day of incubation, respectively. The isolates FAs 1, FAs 4 and FAs 9 were identified as Gram negative non endospore forming rods. In future, these novel isolates possess a great potential in biotechnology field, as bioremediation of arsenic contaminated soil and water can be done by employing arsenic accumulating bacteria which is an eco-friendly and cost effective method

Process optimization for enhanced production of cellulases form locally isolated fungal strain by submerged fermentation

Cellulase has myriad applications in various sectors like pharmaceuticals, textile, detergents, animal feed and bioethanol production, etc. The current study focuses on the isolation, screening and optimization of fungal strain through one factor at a time technique for enhanced cellulase production.  In current study sixteen different fungal cultures were isolated and the culture which quantitatively exhibits higher titers of cellulase activity was identified both morphologically and molecularly by 18S rDNA and designated as Aspergillus niger ABT11. Different parameters like fermentation medium, volume, temperature, pH and nutritional components were optimized. The highest CMCase and FPase activities  was achieved in 100ml of M5 medium in the presence of 1% lactose and sodium nitrate at 30 oC, pH5 after 72 hours. The result revealed A. niger can be a potential candidate for scale up studies

Enhanced production of β- glucosidase by locally isolated fungal strain employing submerged fermentation

β-glucosidase has wide spectrum of biotechnological applications in different industries including food, textile, laundry detergents, pulp and paper, pharmaceutical and biofuel industry. The present investigation related to isolation, screening, and process optimization of fungal strain for enhanced production of β-glucosidase (BGL). For this purpose, different fungal stains were isolated from different sources including soil, fruits, bark of tree as well as from the compost. The screening of fungal strain for BGL production was carried out via submerged fermentation. All the tested strains were identified on the basis of micro and macroscopic features. The fungal strain having greater ability for BGL synthesis among tested ones was identified as Aspergillus niger and given the code SBT-15. The process parameter including fermentation media, temperature, pH, rate of fermentation, carbon and nitrogen sources, volume of media were optimized. Five different fermentation media were evaluated M3 medium gave maximum production. The optimal conditions for BGL production was 72 hours of incubation at 40°C, pH 6 and 50 ml fermentation medium. Glucose (1%) and ammonium sulphate (3%) were optimized as best carbon and nitrogen sources, respectively

Association between TSH status and prevalence of miscarriages and stillbirth

Thyroid hormones play a significant role in normal human body growth. Abnormalities in thyroid stimulating hormone (TSH) levels can result in pregnancy loss due to miscarriages and intrauterine death (IUD). The objective of the study was to assess the levels of association of thyroid stimulating hormone with miscarriages and IUD. The descriptive study involving 110 samples between 18-40 years of age fulfilling inclusion criteria were sampled for TSH testing (2ml blood) after attaining their written informed consent. The mean age of participants was 29.49±4.26 year. The prevalence of hypothyroidism and hyperthyroidism was 3.64% and 2.73%, respectively. Complications like gestational hypertension, depression and oligomenorrhea were found prevalent in these females. Majority of females were taking high/low iodine than recommended iodine level (150mcg). This work shows that there is a significant association between pregnancy loss and disturbed TSH levels among pregnant females

Bioethanol production from urban cellulosic waste employing Alcaligenes faecalis HI-1 isolated from gut of termite Heterotermes indicola

This study assessed the potential of termite gut inhabiting bacteria towards bioconversion of cellulosic waste into biofuel. Total seven bacterial isolates from the gut of Heterotermes indicola were isolated. Among all the isolates, HI-1 produced the largest zone upon primary screening.  Untreated paper had more cellulose content (73.03%) than acid (0.5%) treated paper that was used as a lignocellulosic substrate for saccharification. Among all the isolates tested, glucose yield (1.08mg/mL) was high for HI-1 isolate. Several factors were considered for optimizing augmented glucose yield (8.57mg/mL) and growth (8.07×108cfu/mL), such as temperature 37°C, pH 4.5, 5% (w/v) substrate concentration, 6 % bacterial inoculum size, agitation 150 rpm with PEG 0.25 % and Ca2+ ions 0.002 g/L. Overall 8-fold increase in glucose yield was achieved. Enzyme activity of HI-1 showed higher endoglucanase 0.29 ± 0.01 (U/mL/min) and exoglucanase 0.15±0.01 (U/mL/min) activity under optimum conditions, mentioned above. temperature 37°C, pH 4.5, substrate concentration 5%, inoculum size 6%, surfactants PEG 0.01%, ions Ca2+(0.002g/L) and agitation (120 rpm). Simultaneous saccharification and fermentation (SSF) of hydrolyzed office paper yielded 5.43mg/mL bioethanol. According to 16S rRNA sequence homology, the bacterial isolate H1 was identified as Alcaligenes faecalis. Bioethanol production from office paper untreated waste proved an effective strategy. Bacteria having natural tendency towards cellulosic waste consumption are promising for bioconversion of cellulosic waste to valuable products

Purification and characterisation of α-amylase produced by mutant strain of <i>Aspergillus oryzae</i> EMS-18

<div><p>α-Amylase produced by a mutant strain of <i>Aspergillus oryzae</i> EMS-18 has been purified to homogeneity as judged by sodium dodecyle sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was purified by using 70% ammonium sulphate precipitation followed by anion exchange chromatography on DEAE-Sephadex column and gel filtration on Sephadex G-100. An enzyme purification factor of 9.5-fold was achieved with a final specific activity of 1987.7 U/mg protein and overall yield of 23.8%. The molecular weight of purified α-amylase was estimated to be 48 kDa by SDS-PAGE. The purified enzyme revealed an optimum assay temperature and pH 40°C and 5.0, respectively. Except Ca⁺⁺ all other metal ions such as Mg, Mn, Na, Zn, Ni, Fe, Cu, Co and Ba were found to be inhibitory to enzyme activity.</p></div

Process optimization for hyperproduction of 1,4-alpha-D-glucan glucanohydrolase from locally isolated <em>Bacillus subtilis</em> BBT6 under solid state fermentation

795-800The extracellular enzyme 1,4-alpha-D-glucan glucano-hydrolase (AMY) has wide application in clinical, medicinal and analytical chemistry and are used in industries such as food, textile, paper, etc. The ɑ-amylase (AMY) produced from microbial sources, particularly bacterial is preferred over fungal, because of characteristic advantages, such as rapid growth rate and ability to release proteins into the extracellular medium. The current study deals with the isolation of novel bacterial strain and process optimization of AMY. Primary screening was performed on the basis of starch hydrolysis zone. Secondary screening was carried out using solid state fermentation (SSF). Molecular characterization using16S r RNA gene sequencing technique was performed for the strain showing highest AMY production as compared to other isolates. The selected strain exhibited 91% similarity with the reference strain in the Gene Bank and identified as Bacillus subtilis on the basis of molecular characterization and analytical profile index testing. Different agricultural byproducts such as rice bran, rice husk, wheat bran, potato peel and coconut oil cake was tested. Among all, wheat bran proved to be the best for AMY production. Effect of other variables including incubation time, temperature, pH, inoculum size, carbon and nitrogen sources, surfactant and metal ion have also been investigated. Optimal production of AMY was obtained at 48 h of incubation period, 37°C, pH 7 and inoculum size 1 mL; 1 % ammonium sulphate and peptone as the best inorganic and organic nitrogen sources and 1.5 % starch, 0.2 % SDS found best for optimal production of enzyme

Phytochemical and antimicrobial properties of different plants and in silico investigation of their bioactive compounds in wound healing and rheumatism

In the current study the assessment of the antimicrobial and phytochemical properties of Cassia fistula, Musa paradisiaca, Ficus religiosa and Murraya koenigii plants extracts was carried out. The antibacterial potential of these plants extracts was tested against S. aureus and E. coli. The Cassia fistula and Ficus religiosa leaves showed the larger zone of inhibition in aqueous and butanolic extract respectively against Escherichia coli. Musa paradisiaca and Murraya koenigii leaves showed larger zone of inhibition in ethanolic extract against S. aureus. Qualitative phytochemical analysis showed the presence of alkaloids, flavonoids, phenols, terpenoids, steroids, glycosides, saponins, carbohydrates, proteins and tannins in all extracts while phylobatannins, emodins, anthocyanins and leucoanthocyanins were not present in these extracts. Quantitative phytochemical analysis showed the highest alkaloid content in the Murraya koenigii leaves. Highest tannin content and flavonoid content was found in Ficus religiosa leaves, while highest phenolic content was found in case of Cassia fistula. In addition to this antioxidant potential of all the extracts was determined. Musa paradisiaca leaves showed highest antioxidant potential as compared to other plant extracts. In silico analysis of bioactive components present in plant extracts was performed by molecular docking. The rutin and Glu from Musa paradisiaca and Murraya koenigii respectively, were docked with Glycogen Synthase Kinase 3 beta (1GSK-3beta) protein. Quercetin and rutin from Cassia fistula and Ficus religiosa respectively, were docked with C- reactive protein (CRP). The tested bioactive compounds showed good binding affinity with significant number of hydrogen bonds and can be used as a good alternative of synthetic drugs to treat rheumatism and wounds

Assessment of angiotensin converting enzyme inhibitory activity and quality attributes of yoghurt enriched with Cinnamomum verum, Elettaria cardamomum, Beta vulgaris and Brassica oleracea

The new concept of functional foods has led to the varieties in the production of foods that provide not only basic nutrition, but can also warrant good health and longevity. This study deals with the production and evaluation of fortified yogurts’ with Cinnamomum verum, Elettaria cardamomum, Beta vulgaris and Brassica oleracea. The qualitative and quantitative phytochemical analysis of above mentioned plant extracts before using them into the preparation of functional yoghurt was carried out. The sensory evaluation of enriched yogurts with plant extracts carried out using 9 point hedonic scale. Comparative analysis between enriched yogurts and plain yogurt was carried. The results indicated increase in ash contents, water holding capacity, titratable acidity, total soluble solids, total phenolic content, tannin content, and total flavonoid content in fortified yogurt as compared to plain yogurt. In addition to this fortified yogurts showed greater antioxidant and antibacterial activity in contrast to plain yogurt. However, moisture contents, pH and susceptibility to syneresis of yogurt decreases with the addition of plant extracts. Shelf life of plain and fortified yogurt was determined both at room and refrigerated temperature. The results revealed that shelf life of fortified yogurt was greater as compared to plain yogurt. In silico analysis was carried out by using the galaxy web software. The results indicated that bioactive compounds including ascorbic acid, sinapinic acid, cinnamaldehyde and linalool acetate present in the flavored yogurts binds with angiotensin converting enzyme. All enriched yogurts showed higher anti-Angiotensin converting enzyme activity as compared to plain-yogurt