Biotechnology Research International The latest articles from Hindawi Publishing Corporation © 2015 , Hindawi Publishing Corporation . All rights reserved. Purification, Characterization of L-Methioninase from Candida tropicalis, and Its Application as an Anticancer Tue, 24 Nov 2015 07:17:21 +0000 The aim of the present study is to purify L-methioninase from Candida tropicalis 34.19-fold with 27.98% recovery after ion exchange chromatography followed by gel filtration. The purified enzyme revealed a single band on SDS-PAGE gel with a molecular weight of 46 KDa. Its optimum temperature was 45 to 55 and thermal stability was 55°C for 15 min. The enzyme had optimum pH at 6.5 and stability at a pH range of 5.5 to 7.0 for 24 hr. The maximum activity was observed with substrate concentration of 30 µM and Km was 0.5 mM. The enzyme was strongly inhibited by Cd+2 and Cu+2 while it was enhanced by Na+, Ni+2, and Mg+2 at 10 mM while Ca+2 had slight activation at 20 mM. In addition, the potential application of the L-methioninase as an anticancer agent against various types of tumor cell lines is discussed. Mohsen Helmy Selim, El-Zahraa Karm Eldin, Moataza Mahmoud Saad, El-Sayed Eliwa Mostafa, Yosrea Hassan Shetia, and Amany Ahmed Hassabo Anise Copyright © 2015 Mohsen Helmy Selim et al. All rights reserved. Production of Oxidative and Hydrolytic Enzymes by Coprinus cinereus (Schaeff.) Gray from Sisal Wastes Supplemented with Cow Dung Manure Thu, 19 Nov 2015 17:19:43 +0000 The activity of oxidative and hydrolytic enzymes of the edible and medicinal white rot fungi Coprinus cinereus (Schaeff.) Gray mushroom was observed during mycelia growth and fruiting body development in solid substrate fermentation using sisal waste fractions amended with cow dung manure as supplement. Laccase had the highest titre value among the five detected enzymes. Its activity was higher during mycelia growth compared to fruiting phase, with 10% supplemented substrate formulation unmixed sisal leaf decortication residues [abbreviated SL : SB (100 : 0)] displaying the highest activity of  Ug−1. Lignin peroxidase (LiP) exhibited a characteristic wave-like pattern with the highest peaks found either during full mycelia colonization or soon after first flush harvest; the highest activity of  Ug−1 was observed on unsupplemented SL : SB (100 : 0) substrate formulation during mycelia colonization. For hydrolytic enzymes, the highest carboxymethyl cellulase (CMCase) activity of  Ug−1 was observed on 20% supplemented SL : SB (0 : 100) after first flush; that of pectinase ( Ug−1) was revealed after third flush on 10% supplemented SL : SB (0 : 100) substrate formulation while 10% supplemented SL : SB (25 : 75) exhibited the highest xylanase activity ( Ug−1) after first flush. These findings show that the activities of both oxidative and hydrolytic enzymes were regulated in line with developmental phase of growth of Coprinus cinereus. Prosper Raymond, Anthony Manoni Mshandete, and Amelia Kajumulo Kivaisi Copyright © 2015 Prosper Raymond et al. All rights reserved. Technical and Socioeconomic Potential of Biogas from Cassava Waste in Ghana Wed, 18 Nov 2015 06:35:40 +0000 This study analyses technical potential and ex ante socioeconomic impacts of biogas production using cassava waste from agroprocessing plants. An analysis was performed for two biodigesters in two cassava processing communities in Ghana. The results showed that the two communities generate an excess of 4,500 tonnes of cassava peels per year. Using approximately 5% of the peels generated and livestock manure as inoculum can generate approximately 75,000 m3 of gas with an estimated 60% methane content from two separate plants of capacities 500 m3 and 300 m3 in the two communities. If used internally as process fuel, the potential gas available could replace over 300 tonnes of firewood per year for cassava processing. The displacement of firewood with gas could have environmental, economic, and social benefits in creating sustainable development. With a 10 percent discount rate, an assumed 20-year biodigester will have a Net Present Value of approximately US$ 148,000, 7-year Payback Period, and an Internal Rate of Return of 18.7%. The project will create 10 full-time unskilled labour positions during the investment year and 4 positions during operation years. Francis Kemausuor, Ahmad Addo, and Lawrence Darkwah Copyright © 2015 Francis Kemausuor et al. All rights reserved. Metagenomics: Retrospect and Prospects in High Throughput Age Tue, 17 Nov 2015 09:44:45 +0000 In recent years, metagenomics has emerged as a powerful tool for mining of hidden microbial treasure in a culture independent manner. In the last two decades, metagenomics has been applied extensively to exploit concealed potential of microbial communities from almost all sorts of habitats. A brief historic progress made over the period is discussed in terms of origin of metagenomics to its current state and also the discovery of novel biological functions of commercial importance from metagenomes of diverse habitats. The present review also highlights the paradigm shift of metagenomics from basic study of community composition to insight into the microbial community dynamics for harnessing the full potential of uncultured microbes with more emphasis on the implication of breakthrough developments, namely, Next Generation Sequencing, advanced bioinformatics tools, and systems biology. Satish Kumar, Kishore Kumar Krishnani, Bharat Bhushan, and Manoj Pandit Brahmane Copyright © 2015 Satish Kumar et al. All rights reserved. Mass and Energy Balances of Dry Thermophilic Anaerobic Digestion Treating Swine Manure Mixed with Rice Straw Mon, 02 Nov 2015 13:14:35 +0000 To evaluate the feasibility of swine manure treatment by a proposed Dry Thermophilic Anaerobic Digestion (DT-AD) system, we evaluated the methane yield of swine manure treated using a DT-AD method with rice straw under different C/N ratios and solid retention time (SRT) and calculated the mass and energy balances when the DT-AD system is used for swine manure treatment from a model farm with 1000 pigs and the digested residue is used for forage rice production. A traditional swine manure treatment Oxidation Ditch system was used as the study control. The results suggest that methane yield using the proposed DT-AD system increased with a higher C/N ratio and shorter SRT. Correspondently, for the DT-AD system running with SRT of 80 days, the net energy yields for all treatments were negative, due to low biogas production and high heat loss of digestion tank. However, the biogas yield increased when the SRT was shortened to 40 days, and the generated energy was greater than consumed energy when C/N ratio was and . The results suggest that with the correct optimization of C/N ratio and SRT, the proposed DT-AD system, followed by using digestate for forage rice production, can attain energy self-sufficiency. Sheng Zhou, Jining Zhang, Guoyan Zou, Shohei Riya, and Masaaki Hosomi Copyright © 2015 Sheng Zhou et al. All rights reserved. Extracellular α-Galactosidase from Trichoderma sp. (WF-3): Optimization of Enzyme Production and Biochemical Characterization Mon, 02 Nov 2015 06:52:26 +0000 Trichoderma spp. have been reported earlier for their excellent capacity of secreting extracellular α-galactosidase. This communication focuses on the optimization of culture conditions for optimal production of enzyme and its characterization. The evaluation of the effects of different enzyme assay parameters such as stability, pH, temperature, substrate concentrations, and incubation time on enzyme activity has been made. The most suitable buffer for enzyme assay was found to be citrate phosphate buffer (50 mM, pH 6.0) for optimal enzyme activity. This enzyme was fairly stable at higher temperature as it exhibited 72% activity at 60°C. The enzyme when incubated at room temperature up to two hours did not show any significant loss in activity. It followed Michaelis-Menten curve and showed direct relationship with varying substrate concentrations. Higher substrate concentration was not inhibitory to enzyme activity. The apparent Michaelis-Menten constant (), maximum rate of reaction (), , and catalytic efficiency values for this enzyme were calculated from the Lineweaver-Burk double reciprocal plot and were found to be 0.5 mM, 10 mM/s, 1.30 U mg−1, and 2.33 U mg−1 mM−1, respectively. This information would be helpful in understanding the biophysical and biochemical characteristics of extracellular α-galactosidase from other microbial sources. Aishwarya Singh Chauhan, Arunesh Kumar, Nikhat J. Siddiqi, and B. Sharma Copyright © 2015 Aishwarya Singh Chauhan et al. All rights reserved. Effect of Nitrite and Nitrate Concentrations on the Performance of AFB-MFC Enriched with High-Strength Synthetic Wastewater Thu, 01 Oct 2015 06:49:50 +0000 In order to study the effect of nitrite and nitrate on the performance of microbial fuel cell, a system combining an anaerobic fluidized bed (AFB) and a microbial fuel cell (MFC) was employed for high-strength nitrogen-containing synthetic wastewater treatment. Before this study, the AFB-MFC had been used to treat high-strength organic wastewater for about one year in a continuous flow mode. The results showed that when the concentrations of nitrite nitrogen and nitrate nitrogen were increased from 1700 mg/L to 4045 mg/L and 545 mg/L to 1427 mg/L, respectively, the nitrite nitrogen and nitrate nitrogen removal efficiencies were both above 99%; the COD removal efficiency went up from 60.00% to 88.95%; the voltage was about 375 ± 15 mV while the power density was at 70 ± 5 mW/m2. However, when the concentrations of nitrite nitrogen and nitrate nitrogen were above 4045 mg/L and 1427 mg/L, respectively, the removal of nitrite nitrogen, nitrate nitrogen, COD, voltage, and power density were decreased to be 86%, 88%, 77%, 180 mV, and 17 mW/m2 when nitrite nitrogen and nitrate nitrogen were increased to 4265 mg/L and 1661 mg/L. In addition, the composition of biogas generated in the anode chamber was analyzed by a gas chromatograph. Nitrogen gas, methane, and carbon dioxide were obtained. The results indicated that denitrification happened in anode chamber. Jian-sheng Huang, Ping Yang, Chong-ming Li, Yong Guo, Bo Lai, Ye Wang, Li Feng, and Yun Zhang Copyright © 2015 Jian-sheng Huang et al. All rights reserved. Establishment of MDCK Stable Cell Lines Expressing TMPRSS2 and MSPL and Their Applications in Propagating Influenza Vaccine Viruses in Absence of Exogenous Trypsin Mon, 30 Mar 2015 11:38:10 +0000 We established two Madin-Darby canine kidney (MDCK) cell lines stably expressing human airway transmembrane protease: transmembrane protease, serine 2 (TMPRSS2) and mosaic serine protease large form (MSPL) which support multicycle growth of two H5 highly pathogenic avian influenza viruses (HPAIV) recombinant vaccines (Re-5 and Re-6) and an H9 avian influenza virus (AIV) recombinant vaccine (Re-9) in the absence of trypsin. Data showed that the cell lines stably expressed TMPRSS2 and MSPL after 20 serial passages. Both MDCK-TMPRSS2 and MDCK-MSPL could proteolytically cleave the HA of Re-5, Re-6, and Re-9 and supported high-titer growth of the vaccine without exogenous trypsin. Re-5, Re-6, and Re-9 efficiently infected and replicated within MDCK-TMPRSS2 and MDCK-MSPL cells and viral titer were comparable to the virus grown in MDCK cells with TPCK-trypsin. Thus, our results indicate a potential application for these cell lines in cell-based influenza vaccine production and may serve as a useful tool for HA proteolytic cleavage-related studies. Zhiyuan Wen, Chao Wu, Weiye Chen, Xianying Zeng, Jianzhong Shi, Jinying Ge, Hualan Chen, and Zhigao Bu Copyright © 2015 Zhiyuan Wen et al. All rights reserved. In Silico Analysis of Bioethanol Overproduction by Genetically Modified Microorganisms in Coculture Fermentation Mon, 16 Feb 2015 11:03:03 +0000 Lignocellulosic biomass is an attractive sustainable carbon source for fermentative production of bioethanol. In this context, use of microbial consortia consisting of substrate-selective microbes is advantageous as it eliminates the negative impacts of glucose catabolite repression. In this study, a detailed in silico analysis of bioethanol production from glucose-xylose mixtures of various compositions by coculture fermentation of xylose-selective Escherichia coli strain ZSC113 and glucose-selective wild-type Saccharomyces cerevisiae is presented. Dynamic flux balance models based on available genome-scale metabolic networks of the microorganisms have been used to analyze bioethanol production and the maximization of ethanol productivity is addressed by computing optimal aerobic-anaerobic switching times. A set of genetic engineering strategies for ethanol overproduction by E. coli strain ZSC113 have been evaluated for their efficiency in the context of batch coculture process. Finally, simulations are carried out to determine the pairs of genetically modified E. coli strain ZSC113 and S. cerevisiae that significantly enhance ethanol productivity in batch coculture fermentation. Lisha K. Parambil and Debasis Sarkar Copyright © 2015 Lisha K. Parambil and Debasis Sarkar. All rights reserved. Application of Two-Stage Cultivation for Exploring the Nutritional Requirements for Sporulation of Three Biocontrol Fungi Thu, 29 Jan 2015 16:46:02 +0000 Fungicide was an important part in mycopesticides, which play an important role in pest management, while their mass production and commercialization faced problem. We found that the nutrition for mycelia growth and sporulation differences a lot. So, we developed “two-step method” to define the nutrition for sporulation in this paper. The results indicated that the novel method led to a great increase of spore yields for Beauveria bassiana (IBC1201), Lecanicillium lecanii (CA-1-G), and Pochonia chlamydosporia (HSY-12-14), respectively, of about 100, 2, and 16 times and, also reduced the cycle of mass production to 1/3 compared with common time for culturing. Li Gao Copyright © 2015 Li Gao. All rights reserved. Production and Multiplication of Native Compost Fungal Activator by Using Different Substrates and Its Influence on Growth and Development of Capsicum chinensis Jacq. “Bhut Jolokia” Tue, 06 Jan 2015 12:37:23 +0000 In vitro experiment was carried out to see the effect of saw dusts of Pinus kesiya, Shorea robusta, and Callicarpa arborea on Trichoderma harzianum, isolate TH-13 mass production, along with its biotization effect on Capsicum chinensis Jacq. “Bhut Jolokia.” Early mycelium initiation (2 days) occurred in S. robusta followed by P. kesiya and C. arborea (3.5 days). The sporulation was observed earlier in S. robusta (100% after 6 days) than P. kesiya (33.4% after 8 days) and C. arborea (16.7% after 9 days) but no sporulation was observed in control. The complete sporulation was also earlier in S. robusta (100% after 10 days) than P. kesiya (33.4% after 15 days) and C. arborea (16.4% after 18 days). Higher conidial yield was also in S. robusta than P. kesiya and C. arborea , respectively. The increase in height (60–70 cm), number of leaves (600–650), and yield of chili (120–150 fruits) were also more in inoculated C. chinensis seedlings than control. It is concluded that S. robusta saw dust is the best substrate for mass production of compost fungal activator and can be used in nursery practices for quality stock production of various crops/plantations. Vipin Parkash and Ankur Jyoti Saikia Copyright © 2015 Vipin Parkash and Ankur Jyoti Saikia. All rights reserved. Enhanced Production of Ligninolytic Enzymes by a Mushroom Stereum ostrea Tue, 30 Dec 2014 12:11:53 +0000 The white rot fungi Stereum ostrea displayed a wide diversity in their response to supplemented inducers, surfactants, and copper sulphate in solid state fermentation. Among the inducers tested, 0.02% veratryl alcohol increased the ligninolytic enzyme production to a significant extent. The addition of copper sulphate at 300 μM concentration has a positive effect on laccase production increasing its activity by 2 times compared to control. Among the surfactants, Tween 20, Tween 80, and Triton X 100, tested in the studies, Tween 80 stimulated the production of ligninolytic enzymes. Biosorption of dyes was carried out by using two lignocellulosic wastes, rice bran and wheat bran, in 50 ppm of remazol brilliant blue and remazol brilliant violet 5R dyes. These dye adsorbed lignocelluloses were then utilized for the production of ligninolytic enzymes in solid state mode. The two dye adsorbed lignocelluloses enhanced the production of laccase and manganese peroxidase but not lignin peroxidase. K. Y. Usha, K. Praveen, and B. Rajasekhar Reddy Copyright © 2014 K. Y. Usha et al. All rights reserved. A Metabonomic Study of the Effect of Methanol Extract of Ginger on Raji Cells Using 1HNMR Spectroscopy Sun, 28 Dec 2014 07:35:05 +0000 Cancer is currently a major international health problem. The development of resistance to chemotherapy has resulted in the search for herbal drugs. Ginger is a medicinal plant with several clinical applications. Metabolomics is a simultaneous detection of all the metabolites by use of 1HNMR or mass spectroscopy and interpretation by modeling software. The purpose of this study was to detect the altered metabolites of Raji cells in the presence of ginger extract in vitro. Cells were cultured in the presence and absence of methanolic ginger extract in RPMI medium. IC50 determined by MTT and lipophilic and hydrophilic extracts were prepared from control and treated groups which were analyzed by 1HNMR. The IC50 was 1000 μg/mL. Modeling of spectra was carried out on the two groups using OSC-PLS with MATLAB software and the main metabolites detected. Further analysis was carried out using MetaboAnalyst database. The main metabolic pathways affected by the ginger extract were detected. Ginger extract was seen to effect the protein biosynthesis, amino acid, and carbohydrate metabolism and had a strong cytotoxic effect on Raji cells in vitro. N. Parvizzadeh, S. Sadeghi, S. Irani, A. Iravani, Z. Kalayee, N. A. Rahimi, M. Azadi, and Z. Zamani Copyright © 2014 N. Parvizzadeh et al. All rights reserved. Assessing the Effect of Composting Cassava Peel Based Substrates on the Yield, Nutritional Quality, and Physical Characteristics of Pleurotus ostreatus (Jacq. ex Fr.) Kummer Wed, 17 Dec 2014 08:37:19 +0000 Cassava peel based substrate formulations as an alternative substrate were used to grow mushrooms. The effect of two compost heights, three composting periods on the mycelia growth, physical characteristics, yield, and nutritional qualities of Pleurotus ostreatus (Jacq. ex Fr.) Kummer was studied. Mean mycelia growth of 16.2 cm after a period of seven (7) weeks was the best for 1.5 m compost height. Cap diameter and stipe length differed significantly () with the compost heights (0.8 m and 1.5 m). The yield on compost height of 1.5 m, composted for 5 days, differed significantly () from that of 0.8 m and gave increasing yields as follows: cassava peels and manure, cassava peels only, cassava peels and corn cobs (1 : 1 ratio), and cassava peels and corn cobs (1 : 1 ratio) with chicken manure. Composting periods (3 and 7 days) gave varying yields depending on the compost height. Based on the findings an interaction of 1.5 m compost height and 5 days composting period on cassava peels and corncobs (1 : 1 ratio) with chicken manure produced the best results. The nutritional quality of the mushrooms also differed significantly (), indicating that cassava peels could be used as a possible substrate in cultivation of mushroom. N. K. Kortei, V. P. Dzogbefia, and M. Obodai Copyright © 2014 N. K. Kortei et al. All rights reserved. Biotransformation of Indigo Pigment by Indigenously Isolated Pseudomonas sp. HAV-1 and Assessment of Its Antioxidant Property Mon, 17 Nov 2014 06:53:44 +0000 Chemical synthesis of indigo poses harsh environmental hazards and adverse human health effects. This necessitates an environment-friendly and producer-friendly approach for indigo production. The present study was thus significant as it reports an indigenously isolated potential indigo pigment producing culture identified as Pseudomonas sp. HAV-1 with noteworthy antioxidant property. The bioindigo pigment was characterized using various analytical techniques. The pigment production was enhanced from 412 μg mL−1 to 700 μg mL−1 by optimizing the growth parameters. Furthermore, the antioxidant property of indigo pigment is hitherto unexplored. This property can significantly append to its therapeutic potential. The bioindigo pigment produced by Pseudomonas sp. HAV-1 depicted 2.2 μM ascorbic acid equivalent antioxidant property. More to the point, the present work addresses a footstep towards green production of indigo. Aditi Dua, Kishor Chauhan, and Hilor Pathak Copyright © 2014 Aditi Dua et al. All rights reserved. Ethanol Production from Extruded Thermoplastic Maize Meal by High Gravity Fermentation with Zymomonas mobilis Mon, 03 Nov 2014 07:01:48 +0000 A comparative study of extruded and ground maize meals as raw materials for the production of regular (12°P) and high gravity (20°P) worts was devised. Extruded water solubility index (WSI) was higher (9.8 percentage units) and crude fat was lower (2.64 percentage units) compared to ground maize. Free-amino nitrogen compounds (FAN), pH, and glucose were evaluated in regular and high gravity worts produced from ground or extruded maize. Extrusion improved glucose content and ethanol yield. In 20°P mashes, extrusion is enhanced by 2.14% initial glucose compared with regular ground mashes. The 12°P and 20°P extruded treatments averaged 12.2% and 8.4% higher ethanol, respectively, compared to the uncooked counterpart. The 20°P worts fermented with Zymomonas mobilis produced 9.56% more ethanol than the 12°P counterpart. The results show that the combination of extrusion and fermentation of 20°P worts improved ethanol yield per kg flour until 20.93%. This pretreatment stimulates Z. mobilis fermentation efficiency. Mayeli Peralta-Contreras, Edna Aguilar-Zamarripa, Esther Pérez-Carrillo, Erandi Escamilla-García, and Sergio Othon Serna-Saldívar Copyright © 2014 Mayeli Peralta-Contreras et al. All rights reserved. Comparative Studies of Oleaginous Fungal Strains (Mucor circinelloides and Trichoderma reesei) for Effective Wastewater Treatment and Bio-Oil Production Sun, 02 Nov 2014 08:00:36 +0000 Biological wastewater treatment typically requires the use of bacteria for degradation of carbonaceous and nitrogenous compounds present in wastewater. The high lipid containing biomass can be used to extract oil and the contents can be termed as bio-oil (or biodiesel or myco-diesel after transesterification). The separate experiments were conducted on actual wastewater samples with 5% v/v inoculum of Mucor circinelloides MTCC1297 and Trichoderma reesei NCIM992 strains. The observed reductions in chemical oxygen demand (COD) were 88.72% and 86.75% in 96 hrs and the observed substrate based biomass yields were 0.21 mg VSS/mg COD and 0.22 mg VSS/mg COD for M. circinelloides reactor and for T. reesei reactor, respectively. The resulted bio-oil production from wastewater treatment by M. circinelloides and T. reesei reactors was 142.2 mg/L and 74.1 mg/L, whereas biomass containing bio-oil contents (%w/w) were 22.11% and 9.82%, respectively. In this experiment, the fungal wastewater treatment was also compared with conventional bacterial process with respect to specific growth rate, biomass production, and oil content. This study suggests that wastewater can be used as a potential feedstock for bio-oil production with the use of oleaginous fungal strains and which could be a possible route of waste to energy. Anshuman Bhanja, Gauri Minde, Sandip Magdum, and V. Kalyanraman Copyright © 2014 Anshuman Bhanja et al. All rights reserved. Overcome of Carbon Catabolite Repression of Bioinsecticides Production by Sporeless Bacillus thuringiensis through Adequate Fermentation Technology Wed, 17 Sep 2014 09:59:47 +0000 The overcoming of catabolite repression, in bioinsecticides production by sporeless Bacillus thuringiensis strain S22 was investigated into fully controlled 3 L fermenter, using glucose based medium. When applying adequate oxygen profile throughout the fermentation period (75% oxygen saturation), it was possible to partially overcome the catabolite repression, normally occurring at high initial glucose concentrations (30 and 40 g/L glucose). Moreover, toxin production yield by sporeless strain S22 was markedly improved by the adoption of the fed-batch intermittent cultures technology. With 22.5 g/L glucose used into culture medium, toxin production was improved by about 36% when applying fed-batch culture compared to one batch. Consequently, the proposed fed-batch strategy was efficient for the overcome of the carbon catabolite repression. So, it was possible to overproduce insecticidal crystal proteins into highly concentrated medium. Saoussen Ben Khedher, Samir Jaoua, and Nabil Zouari Copyright © 2014 Saoussen Ben Khedher et al. All rights reserved. Antidepressant Fluoxetine Modulates the In Vitro Inhibitory Activity of Buffalo Brain Cystatin: A Thermodynamic Study Using UV and Fluorescence Techniques Thu, 24 Jul 2014 08:59:07 +0000 Cystatins constitute a superfamily of homologous proteins. The major role of cystatins is to regulate the unwanted proteolysis and to protect the organism against endogenous proteases released from lysosomes, invading microorganisms and parasites that use cysteine proteases to enter the body. Imbalance in regulation of proteolytic activity may lead to a wide range of human diseases. An enormous progress has been made in understanding of protein degradation process under normal and pathological conditions; infact proteases are now clearly viewed as important drug targets. Fluoxetine a selective serotonin reuptake inhibitor (SSRI) is an antidepressant. It is used to treat major depressive disorders. In the present study binding of fluoxetine to cystatin was studied by UV and fluorescence quenching technique. Intrinsic fluorescence of fluoxetine complexed with purified buffalo brain cystatin (BC) was measured by selectively exciting the tryptophan residues. Gradual quenching was observed on complex formation. When cystatin was added to fluoxetine solutions at a molar ratio of 1 : 0.5, it not only quenched more than half of its fluorescence but also reduced the activity of cystatin. Stern-Volmer plots obtained from experiments carried out at 25°C showed the quenching of fluorescence to be a collisional phenomenon. Our results suggest the prime binding site for fluoxetine on BC to be at or near tryptophan residues. Fluoxetine quenched the fluorescence by a static process, which specifically indicates the formation of a complex. Fakhra Amin and Bilqees Bano Copyright © 2014 Fakhra Amin and Bilqees Bano. All rights reserved. Isolation and Characterization of Bacteriophages Infecting Nocardioforms in Wastewater Treatment Plant Tue, 22 Jul 2014 07:33:42 +0000 Activated sludge plants (ASP) are associated with the stable foaming problem worldwide. Apart from the physical and chemical treatment methods, biological treatment method has been least explored and may prove to be a novel and ecofriendly approach to tackle the problem of stable foam formation. In ASP Nocardia species are commonly found and are one of the major causes for forming sticky and stable foam. This study describes the isolation and characterization of three Nocardia bacteriophages NOC1, NOC2, and NOC3 for the control of Nocardia species. The bacteriophages isolated in this study have shown promising results in controlling foam producing bacterial growth under laboratory conditions, suggesting that it may prove useful in the field as an alternative biocontrol agent to reduce the foaming problem. To the best of our knowledge to date no work has been published from India related to biological approach for the control of foaming. Krishna Khairnar, Preeti Pal, Rajshree H. Chandekar, and Waman N. Paunikar Copyright © 2014 Krishna Khairnar et al. All rights reserved. Optimization of Medium Composition for the Production of Neomycin by Streptomyces fradiae NCIM 2418 in Solid State Fermentation Mon, 09 Jun 2014 09:18:59 +0000 Neomycin production of Streptomyces fradiae NCIM 2418 was optimized by using response surface methodology (RSM), which is powerful mathematical approach comprehensively applied in the optimization of solid state fermentation processes. In the first step of optimization, with Placket-Burman design, ammonium chloride, sodium nitrate, L-histidine, and ammonium nitrate were established to be the crucial nutritional factors affecting neomycin production significantly. In the second step, a 24 full factorial central composite design and RSM were applied to determine the optimal concentration of significant variable. A second-order polynomial was determined by the multiple regression analysis of the experimental data. The optimum values for the important nutrients for the maximum were obtained as follows: ammonium chloride 2.00%, sodium nitrate 1.50%, L-histidine 0.250%, and ammonium nitrate 0.250% with a predicted value of maximum neomycin production of 20,000 g kg−1 dry coconut oil cake. Under the optimal condition, the practical neomycin production was 19,642 g kg−1 dry coconut oil cake. The determination coefficient (R2) was 0.9232, which ensures an acceptable admissibility of the model. B. M. Vastrad and S. E. Neelagund Copyright © 2014 B. M. Vastrad and S. E. Neelagund. All rights reserved. Fermented Fruits and Vegetables of Asia: A Potential Source of Probiotics Wed, 28 May 2014 09:56:43 +0000 As world population increases, lactic acid fermentation is expected to become an important role in preserving fresh vegetables, fruits, and other food items for feeding humanity in developing countries. However, several fermented fruits and vegetables products (Sauerkraut, Kimchi, Gundruk, Khalpi, Sinki, etc.) have a long history in human nutrition from ancient ages and are associated with the several social aspects of different communities. Among the food items, fruits and vegetables are easily perishable commodities due to their high water activity and nutritive values. These conditions are more critical in tropical and subtropical countries which favour the growth of spoilage causing microorganisms. Lactic acid fermentation increases shelf life of fruits and vegetables and also enhances several beneficial properties, including nutritive value and flavours, and reduces toxicity. Fermented fruits and vegetables can be used as a potential source of probiotics as they harbour several lactic acid bacteria such as Lactobacillus plantarum, L. pentosus, L. brevis, L. acidophilus, L. fermentum, Leuconostoc fallax, and L. mesenteroides. As a whole, the traditionally fermented fruits and vegetables not only serve as food supplements but also attribute towards health benefits. This review aims to describe some important Asian fermented fruits and vegetables and their significance as a potential source of probiotics. Manas Ranjan Swain, Marimuthu Anandharaj, Ramesh Chandra Ray, and Rizwana Parveen Rani Copyright © 2014 Manas Ranjan Swain et al. All rights reserved. Characterization of Modified Magnetite Nanoparticles for Albumin Immobilization Thu, 22 May 2014 12:12:54 +0000 Magnetite Fe3O4 nanoparticles (NPs) were prepared by chemical coprecipitation method. Silica-coated magnetite NPs were prepared by sol-gel reaction, subsequently coated with 3-aminopropyltriethoxysilane (APTES) via silanization reaction, and then were activated with 2,4,6-trichloro-1,3,5-triazine (TCT) and covalently immobilized with bovine serum albumin (BSA). The size and structure of the particles were characterized by transmission electron microscopy (TEM), X-ray powder diffraction (XRD), and dynamic light scattering (DLS) techniques. The immobilization was confirmed by Fourier transform infrared spectroscopy (FT-IR). XRD analysis showed that the binding process has not done any phase change to Fe3O4. The immobilization time for this process was 4 h and the amount of immobilized BSA for the initial value of 1.05 mg BSA was about 120 mg/gr nanoparticles. Also, the influences of three different buffer solutions and ionic strength on covalent immobilization were evaluated. A. K. Bordbar, A. A. Rastegari, R. Amiri, E. Ranjbakhsh, M. Abbasi, and A. R. Khosropour Copyright © 2014 A. K. Bordbar et al. All rights reserved. Algae Oil: A Sustainable Renewable Fuel of Future Mon, 05 May 2014 12:55:25 +0000 A nonrenewable fuel like petroleum has been used from centuries and its usage has kept on increasing day by day. This also contributes to increased production of greenhouse gases contributing towards global issues like global warming. In order to meet environmental and economic sustainability, renewable, carbon neutral transport fuels are necessary. To meet these demands microalgae are the key source for production of biodiesel. These microalgae do produce oil from sunlight like plants but in a much more efficient manner. Biodiesel provides more environmental benefits, and being a renewable resource it has gained lot of attraction. However, the main obstacle to commercialization of biodiesel is its cost and feasibility. Biodiesel is usually used by blending with petro diesel, but it can also be used in pure form. Biodiesel is a sustainable fuel, as it is available throughout the year and can run any engine. It will satisfy the needs of the future generation to come. It will meet the demands of the future generation to come. Monford Paul Abishek, Jay Patel, and Anand Prem Rajan Copyright © 2014 Monford Paul Abishek et al. All rights reserved. Parametric Optimization of Cultural Conditions for Carboxymethyl Cellulase Production Using Pretreated Rice Straw by Bacillus sp. 313SI under Stationary and Shaking Conditions Tue, 29 Apr 2014 07:01:06 +0000 Carboxymethyl cellulase (CMCase) provides a key opportunity for achieving tremendous benefits of utilizing rice straw as cellulosic biomass. Out of total 80 microbial isolates from different ecological niches one bacterial strain, identified as Bacillus sp. 313SI, was selected for CMCase production under stationary as well as shaking conditions of growth. During two-stage pretreatment, rice straw was first treated with 0.5 M KOH to remove lignin followed by treatment with 0.1 N H2SO4 for removal of hemicellulose. The maximum carboxymethyl cellulase activity of 3.08 U/mL was obtained using 1% (w/v) pretreated rice straw with 1% (v/v) inoculum, pH 8.0 at 35°C after 60 h of growth under stationary conditions, while the same was obtained as 4.15 U/mL using 0.75% (w/v) pretreated substrate with 0.4% (v/v) inoculum, pH 8.0 at 30°C, under shaking conditions of growth for 48 h. For maximum titre of CMCase carboxymethyl cellulose was optimized as the best carbon source under both cultural conditions while ammonium sulphate and ammonium nitrate were optimized as the best nitrogen sources under stationary and shaking conditions, respectively. The present study provides the useful data about the optimized conditions for CMCase production by Bacillus sp. 313SI from pretreated rice straw. Varsha Goyal, Arpana Mittal, Anish Kumari Bhuwal, Gulab Singh, Anita Yadav, and Neeraj Kumar Aggarwal Copyright © 2014 Varsha Goyal et al. All rights reserved. Cloning, Sequencing, and In Silico Analysis of -Propeller Phytase Bacillus licheniformis Strain PB-13 Thu, 24 Apr 2014 10:08:09 +0000 β-Propeller phytases (BPPhy) are widely distributed in nature and play a major role in phytate-phosphorus cycling. In the present study, a BPPhy gene from Bacillus licheniformis strain was expressed in E. coli with a phytase activity of 1.15 U/mL and specific activity of 0.92 U/mg proteins. The expressed enzyme represented a full length ORF “PhyPB13” of 381 amino acid residues and differs by 3 residues from the closest similar existing BPPhy sequences. The PhyPB13 sequence was characterized in silico using various bioinformatic tools to better understand structural, functional, and evolutionary aspects of BPPhy class by multiple sequence alignment and homology search, phylogenetic tree construction, variation in biochemical features, and distribution of motifs and superfamilies. In all sequences, conserved sites were observed toward their N-terminus and C-terminus. Cysteine was not present in the sequence. Overall, three major clusters were observed in phylogenetic tree with variation in biophysical characteristics. A total of 10 motifs were reported with motif “1” observed in all 44 protein sequences and might be used for diversity and expression analysis of BPPhy enzymes. This study revealed important sequence features of BPPhy and pave a way for determining catalytic mechanism and selection of phytase with desirable characteristics. Vinod Kumar, Gopal Singh, Punesh Sangwan, A. K. Verma, and Sanjeev Agrawal Copyright © 2014 Vinod Kumar et al. All rights reserved. Statistical Optimization of Media Components for Production of Fibrinolytic Alkaline Metalloproteases from Xenorhabdus indica KB-3 Wed, 23 Apr 2014 15:03:56 +0000 Xenorhabdus indica KB-3, a well-known protease producer, was isolated from its entomopathogenic nematode symbiont Steinernema thermophilum. Since medium constituents are critical to the protease production, the chemical components of the selected medium (soya casein digest broth) were optimized by rotatable central composite design (RCCD) using response surface methodology (RSM). The effects of all five chemical components (considered as independent variables), namely tryptone, soya peptone, dextrose, NaCl, and dipotassium phosphate, on protease production (dependent variable) were studied, and it was found that tryptone and dextrose had maximum influence on protease production. The protease production was increased significantly by 66.31% under optimal medium conditions (tryptone—5.71, soya peptone—4.9, dextrose—1.45, NaCl—6.08, and dipotassium phosphate—0.47 in g/L). To best of knowledge, there are no reports on optimization of medium component for protease production by X. indica KB-3 using RSM and their application in fibrinolysis. This study will be useful for industrial processes for production of protease enzyme from X. indica KB-3 for its application in the field of agriculture and medicine. Kumar Pranaw, Surender Singh, Debjani Dutta, Surabhi Chaudhuri, Sudershan Ganguly, and Lata Nain Copyright © 2014 Kumar Pranaw et al. All rights reserved. Biocontrol Activity of the Local Strain of Metschnikowia pulcherrima on Different Postharvest Pathogens Thu, 17 Apr 2014 06:37:32 +0000 The strains of the yeast Metschnikowia pulcherrima have strong biocontrol activity against various microorganisms. Biocontrol activity of M. pulcherrima largely depends on its iron immobilizing pigment pulcherrimin. Biocontrol activity of pulcherrimin producing strain, M. pulcherrima UMY15, isolated from local vineyards, was tested on different molds that cause food spoilage. M. pulcherrima UMY15 was a very effective biocontrol agent against Penicillium roqueforti, P. italicum, P. expansum, and Aspergillus oryzae in in-vitro plate tests. However, the inhibitory activity of M. pulcherrima UMY15 was less effective on Fusarium sp. and A. niger species in biocontrol assays. In addition, M. pulcherrima UMY15 strain completely inhibited the germination and mycelia growth of A. oryzae, A. parasiticus, and Fusarium sp. spores on artificial wounds of apples when they coinoculated with M. pulcherrima UMY15. Moreover, when coinoculated, M. pulcherrima UMY15 strain also inhibited the growth of P. roqueforti, P. italicum, P. expansum, A. oryzae, Fusarium sp., and Rhizopus sp. in grape juice, indicating that M. pulcherrima UMY15 can be used as a very effective biocontrol yeast against various species of postharvest pathogens, including   Penicillium, Aspergillus, Fusarium, and Rhizopus. Sezai Türkel, Mihriban Korukluoğlu, and Mümine Yavuz Copyright © 2014 Sezai Türkel et al. All rights reserved. Evaluation of Novel Design Strategies for Developing Zinc Finger Nucleases Tools for Treating Human Diseases Sun, 06 Apr 2014 11:09:24 +0000 Zinc finger nucleases (ZFNs) are associated with cell death and apoptosis by binding at countless undesired locations. This cytotoxicity is associated with the binding ability of engineered zinc finger domains to bind dissimilar DNA sequences with high affinity. In general, binding preferences of transcription factors are associated with significant degenerated diversity and complexity which convolutes the design and engineering of precise DNA binding domains. Evolutionary success of natural zinc finger proteins, however, evinces that nature created specific evolutionary traits and strategies, such as modularity and rank-specific recognition to cope with binding complexity that are critical for creating clinical viable tools to precisely modify the human genome. Our findings indicate preservation of general modularity and significant alteration of the rank-specific binding preferences of the three-finger binding domain of transcription factor SP1 when exchanging amino acids in the 2nd finger. Christian Bach, William Sherman, Jani Pallis, Prabir Patra, and Hassan Bajwa Copyright © 2014 Christian Bach et al. All rights reserved. Characterization of β-Glucosidase Produced by Aspergillus niger under Solid-State Fermentation and Partially Purified Using MANAE-Agarose Tue, 01 Apr 2014 12:29:52 +0000 β-Glucosidase (BGL) is a hydrolytic enzyme with specificity for a wide variety of glycoside substrates, being an enzyme with a large range of biotechnological applications. However, enzyme properties can be different depending both on the microorganism and the cultivation procedure employed. Therefore, in order to explore potential biocatalytical applications of novel enzymes, their characterization is essential. In this work, a BGL synthesized by a selected strain of Aspergillus niger cultivated under solid-state fermentation (SSF) was partially purified and fully characterized in terms of optimum pH, temperature, and thermostability. The single-step purification using MANAE-agarose in a chromatographic column yielded an enzyme solution with specific activity (17.1 IU/mg protein) adequate for the characterization procedures. Electrophoresis SDS-PAGE and size-exclusion chromatography analysis resulted in an estimated molecular mass of 60 kDa. Higher enzyme activities were found in the range between 40 and 65°C and between pH 4 and 5.5, indicating an interesting characteristic for application in the hydrolysis of lignocellulosic biomass for biofuels production. Thermostability studies of purified BGL resulted in half-lives at 37°C of 56.3 h and at 50°C of 5.4 h. These results provide support for further studies of this enzyme towards revealing its potential biotechnological applications. Anderson Baraldo Junior, Diogo G. Borges, Paulo W. Tardioli, and Cristiane S. Farinas Copyright © 2014 Anderson Baraldo Junior et al. All rights reserved.