Research Journal of Biotechnology

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Research Journal of Biotechnology

Transformation of L-Tyrosine to L-Dopa by Acremonium rutilum W.Gams: Effect of Nutritional Parameters

Rathod Vandana and Ramakrishna Krishnaveni

In previous study transformation of L-tyrosine to L-dopa by A.rutilum under submerged fermentation was reported where physical parameters and substrate concentration for the optimization of L-dopa were studied. Present study reports the effect of nutritional parameters on L-dopa yield by A.rutilum. Results showed that 4% glucose showed highest yield of L-dopa (0.89mg/ml) followed by 3% starch (0.85 mg/ml). Egg flakes probably are known to slow down the catecholase activity of tyrosinase thus increasing L-Dopa production proving it to be the best organic nitrogen source with maximum L-Dopa production (1.06mg/ml). At 4 mg/100ml of copper sulphate, the yield of L-Dopa was 1.09 mg/ml. Enriched potato glucose albumen broth (EGAB) proved to be the best media, easier and economical for L-dopa pro­d­uction. Overall process consistency revealed tyr­os­inase activity to be 40U/ml and 1.12mg/ml L-Dopa yield.

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Production of Poly-3-hydroxybutyrate (PHB) from lactose and whey by Bacillus thuringiensis IAM 12077

Srividya Shivakumar

Bacillus thuringiensis IAM 12077, an indigenous soil bacterium, is able to grow in a medium containing 10 g l−1 lactose as a sole source of carbon, giving 3.6 g l−1 biomass yield and poly-3-hydroxybutyrate (PHB) up to 40% of its dry weight in 48 h. The isolate was also able to utilize whole cheese whey and whey supernate and produce 3•9 g l−1 and 3.3 g l-1 PHB respectively by 72 h. Lactose at a concentration of 2 % (w/v) when supplemented with whey decreased the time required for maximum PHB production from 72 h to 48 h. Similarly a combination of lactose (1 %, w/v) supplemented with different concentrations of whey amounting to a final concentration of carbon source in the range from 1 % to 10 %, also showed 2 % (w/v) carbon source to be optimum for supporting maximum PHB production by this strain. Whole whey showed 67.5 % accumulation with 3.9 g/L yield by 72 h and whey supernate showed 62.5 % accumulation with 3.3 g/L PHB yield. B.thuringiensis IAM 12077 is capable of accumulating appreciable levels of PHB from lactose, whey (whole and supernate) and therefore offers much potential for economic production of PHB from such raw materials.

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Development of an Efficient Marker-Assisted Backcrossing Scheme in Rice using Microsatellite Markers

Joshi Raj Kumar

A marker-assisted backcrossing scheme was performed using micro satellite markers in the genetic background of rice to deduce the best possible criteria for performing marker-assisted background selection. 72 polymorphic SSR markers were selected for genotyping 123 BC1 rice plants developed from a cross between two rice cultivars IR64 (recurrent parent) and Tetep (Donor parent with Pi-kh resistance gene). The BC2 generation produced 144 plants and screened with markers not amplified in the BC1 generation. 400 plants were produced in the BC3 generation and 48 plants were selected for recurrent parent genotype recovery. The target marker (polymorphic micro satellite RM206) was transferred efficiently. The recurrent parent genotype recovery averages in three backcrosses indicates that marker-assisted backcrossing produced a genetic gain. Backcross transfer at genotype level showed high accuracy when six markers per chromosomes were used even in a saturated rice genome. Morpho-agronomically, the plants in SC3 criteria (six markers per chromosome) were closer to the recurrent parent than in any other criteria. Thus, it can be concluded that six markers per chromosome can be considered as the best option to perform marker-assisted background selection in three generations.

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In Vitro Anti Bacterial Studies of Gardenia Resinifera Roth. and Gardenia Latifolia Ait.

Jhansi Lakshmi B. and Jaganmohanreddy K.

The aqueous and methanolic extractions of Gardenia resinifera and Gardenia latifolia were evaluated for antibacterial activity against Bacillus cereus, Bacillus megatherium, Staphylococcus aureus, Enterobacter aerogenes and Salmonella paratyphi. The in vitro antibacterial activity was performed by agar well diffusion method. Gram positive bacteria showed more antibacterial activity than Gram negative bacteria. Leaf extracts showed higher activity when compared with the bark extracts of G.resinifera and G.latifolia. Methanolic extractions sho­wed higher activity than aqueous extractions. The DMSO did not exhibit any antibacterial activity against any bacterial organism tested in the present study.

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Thermal Inactivation Kinetics of Opuntia vulgaris xylose isomerase isoenzymes

Sambandam Ravikumar and Kotteazeth Srikumar

The thermal inactivation kinetics of two isoenzyme preparations of the xerophyte Opuntia vulgaris xylose isomerase was investigated in solution. The kinetic parameters were determined from graphical plots by use of least-square method using nonlinear regression analysis. Each of the two isoenzyme exhibited first-order kinetics with different inactivation rates. Inactivation kinetics of this isoenzyme in the presence of metal ions (Mn2+, Co2+ and Mg2+) also exhibited first-order. Heat inactivation data (based on percent residual activity) for this enzyme activity in the presence and absence of metal ions was determined employing the first-order inactivation rate constants and enzyme reaction rates. The results aided thermodynamical interpretation of the relationship between inactivation rates and metal binding. Differences in the inactivation kinetics between the isoenzymes could be explained on the basis of their differences in heat resistance.

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Study on the antimicrobial activity of honey products and some Saudi Folkloric substances

Ghanem Nevine B.

Alcoholic and aqueous extracts from 17 Saudi Arabia folkloric substances were screened for antimicrobial against some tested pathogenic microorganisms by zone of inhibition assay and minimal inhibitory concentration (MIC). Compared to honey and bee wax, ethanolic extract of propolis showed the highest zone of inhibition (23mm) against S. aureus ATCC255923. Aqueous extract of alum (Ming Fan) was highly effective against all tested strains with inhibition zones ranging from 25 to 29 mm followed by Juglans regia (28mm) with S. epidermis ATCC12228, Rhazya stricta (24mm) with Str. pyogenes ATCC19615, and Commiphora myrrha (24mm) with S. aureus ATCC255923. The combined effect of equimixture of ethanolic extracts of propolis and bee wax was more effective than a single extract showing 1.5 fold increase of inhibition zone against S. aureus ATCC255923 and C. albicans NCTC2708. Substances with the most consistent action against microorganisms were tested to determine their minimum inhibitory concentration (MIC). Alum (Ming Fan) was found to have the greatest activity with MIC mean value of 0.29% (w/v) followed by Rhazya stricta, Juglans regia and propolis with mean MIC values of 0.4, 2.66 and 3.75 respectively.

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Effect of Arbuscular Mycorrhizal Fungi on the growth of Ocimum sanctum and Glomalin a soil related protein

Padmavathi T. and Ranjini R.

Ocimum sanctum (tulsi) is a medicinal plant that confers longevity. Its therapeutic use proves miraculously successful. Each part of the plant possesses medicinal properties. Ocimum sanctum is native throughout the World tropics and widespread as a cultivated plant and an escaped weed. It is cultivated for religious and medicinal purposes and for its essential oil. Although it has been well established that mycorrhizae enhance nutrient uptake and crop yield but there is no information on AM fungal interaction with Ocimum sanctum plants in India. One of the strategies in AM research has been to manage the indigenous population of the AM fungi to different agronomic practice. AM fungi were wide spread in their occurrence in the fields of Bangalore. The no. of spores contained in fifty grams of soil ranged from 26-203 in the samples collected from unplanted soils. When ocimum plants were dually inoculated with Glomus intraradices and Azospirillum lipoferum shoot biomass production increased by 98% when compared to untreated plants. The maximum yield of shoot dry mass was obtained when plants were treated with 10g of A.lipoferum and G.intraradices. Root biomass significantly increased when plants were inoculated with 10g of A.lipoferum and AM fungi. Plants inoculated with 10g of A.lipoferum and AM fungi increased significantly root dry mass. Inoculating Ocimum santum with G.intraradices and 15g of A.lipoferum resulted into 79.3% colonization. Plants treated with 10g of A.lipoferum and AM fungi had 143 spores for 50g of soil sample. The dual inoculation increased the spore no. as well as the growth of ocimum plants. Soil organic matter (SOM) is a key component of soil that greatly influences its structure and productivity as well as aggregate stability. High soil aggregate stability translates to less soil erosion and hence lessens the likelihood of non-point-source water pollution. AM fungi increasing soil aggregate stability led to the soil protein called Glomalin produced in abundance by the hyphae of these fungi. Glomalin, a soil related protein, was found in all the treatments with ocimum plants. 278.47 micrograms/ml of glomalin were obtained when plants were treated with 5g of A.lipoferum and AM fungi. An attempt was also made to predict the 3D structure of soil protein glomalin by Swiss Model Server and its multimeric form using the Cluspro server.

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Study of Biosorptive Potential in the Peel of Citris reticulatae, Punica grantum, Daucus carota and Momordica charantia

Ashraf Muhammad Aqeel, Maah Mohd. Jamil, Yusoff Ismail , Mahmood Karamat and Wajid Abdul

Different physiochemical parameters such as color, order, taste, pH, electrical conductivity, total dissolved solids, concentration of cations and anions were determined in the waste water samples that were collected in the vicinity of Kuala Lumpur city. Peel of Citris reticulatae (Orange), Punica Grantum (Pomegranete), Daucus carota (Carrot) and Momordica charantia (Bitter Gourd) were used to study biosorption potential of the dead biomass. Laboratory batch experiments indicated that Citris reticulatae (Orange), Punica Grantum (Pomeg­ranete), Daucus carota (Carrot) and Momordica charantia (Bitter Gourd) had an excellent ability to bind with these anions and cations. The role of cell structure, cell wall, micropores and macropores describe the potential of adsorbent. Batch laboratory time dependency, pH profile and concentration parameter were performed to determine the binding ability of biosorbents for cations and anions. Time dependency indicated that first three samples showed maximum adsorption at 45 min. and Punica granatum at 60 minutes and at pH 12. More amount of biosorbent is used for that water sample which contains maximum amount of cations and anions. These are recommended as good biosorbent due to their effectiveness.

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Bactericidal and Bioactivity guided Fractionation Studies of Mangrove Species Derris Indica and D. Trifoliata

Mitter C. S. and Jadhav B. L.

Bactericidal and bioactivity guided fractionation studies of D. indica and D. trifoliata were carried out. The cold and hot extracts of leaf, bark and tender fruits of these plants were made in petroleum ether, chloroform, ethyl acetate, acetone, ethanol, methanol and water solvent and tested against several human pathogens. Overall better bactericidal activity was registered in D. indica against Staphylococcus aureus, Corynebacterium diphtheriae and Klebseilla pneumoniae. The cold ethyl acetate extracts of bark and tender fruits of D. indica exhibited MIC (Minimum Inhibitory Concentration) up to 5mg/ml against C. diphtheriae and Pseudomonas aeruginosa. Activity guided fractionation of cold bark and tender fruits of ethyl acetate extracts showed activity comparable to streptomycin and ofloxacin in petroleum ether, ethyl acetate and methanol fractions against C. diphtheriae, S. aureus and Ps. aeruginosa.

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Physico-chemical properties of dehydrated olive mill wastewater pellets

Ruiz Celma A., López-Rodríguez F. and Cuadros F.

The current market price of olive mill wastewater might be regarded as negligible. Moreover, its handling often leads to additional costs for the olive oil industry. This way, the pelletizing process for such by-product emerges as a relevant task to achieve a reduction of handling and transport costs as well as to promote new alternatives for its use.The process described in the present paper is the manufacturing of 6 mm diameter pellets from olive mill wastewater samples obtained from sludge processing plants. As a previous step, the concentrated samples were dehydrated to three different values of moisture content (23, 16 and 10 wt% w.b.). A detailed analysis of the following physical-chemical properties of the concentrated olive mill wastewater pellets obtained in the laboratory has been carried out: particle and bulk densities (kg/m3), hardness (N), durability (%) and resistance to wetting. Fluctuations of those parameters caused by the addition of another by-product from the olive oil industry (olive husk, 8 wt% w.b. moisture content) during the manufacturing process are also reported. Pellets obtained exclusively from dehydrated olive mill wastewater, no matter the moisture content of samples, were found to show highest durability (>98.9%) and best results in resistance to wetting. On the other hand, the addition of olive husk during the manufacturing process led to a general tendency governed by an increase in hardness and a decrease in durability. Such behavior was observed for olive husk concentrations up to 75% in weight.

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Impact of Organic Amendment of Substrate on Growth of Agaricus Bisporus (Lange) Sing.

Sharma G.D., Sharan A.K. and Jee Chandrawati

In recent years attempts have been made to amend the nature of the substrate on which A. bisporus can be suitably cultivated. In present study attempts have been made to amend the substrate using neem cake and castor cake at a concentration of 125g, 250g and 375g to monitor the effect. Variable nature of growth pattern has been noticed. It has been found that 375g of neem cake increases the yield after 15 days and 30 days to the extent of 35% and 76%. Castor cake treatment at a concentration of 125g and 250g increases the yield by 61% and 59% after 15 days. Treatment for 30 days with 375g of castor cake exhibits the most suitable result as emergence of sporophore increase by 19% (145) over control (77). An overview of the data suggest reduction in time of emergence of sporophore by five days due to these treatments in natural growing condition and 30 days treatment with 375g of both neem cake and castor cake.

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Plant –Microbe Interaction with Enhanced Bioremediation

Bhatia Divya and Malik Deepak Kumar

Bioremediation of organic pollutants is often a slow and incomplete process, potentially leading to the accumulation of toxic metabolites that can be further introduced into the food chain. Plant-associated bacteria, such as endophytic bacteria (non-pathogenic bacteria that occur naturally in plants) and rhizospheric bacteria (bacteria that live on and near the roots of plants), have been shown to contribute to biodegradation of toxic organic compounds in contaminated soil and could have potential for improving phytoremediation. The combi­nation of bioaugmentation and phytoremed­iation, resulting in rhizoremediation, could solve some of the problems encountered during the application of both individual techniques. The inoculation of pollutant-degrading bacteria on plant seed can be an important additive to improve the efficiency of phyto­remediation or bioaugmentation. Genetic engine­ee­ring of endo­phytic and rhizospheric bacteria for use in plant-associated degradation of toxic compounds in soil is considered one of the most promising new technol­ogies for remediation of contaminated environmental sites. Transgenic plants exhibiting biodegradation capabilities of microorganisms bring the promise of an efficient and environmental-friendly technology for cleaning up polluted soils.

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