ISRN Biotechnology http://www.hindawi.com The latest articles from Hindawi Publishing Corporation © 2014 , Hindawi Publishing Corporation . All rights reserved. Use of Metagenomics and Isolation of Actinobacteria in Brazil’s Atlantic Rainforest Soil for Antimicrobial Prospecting Wed, 12 Mar 2014 06:59:14 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2014/909601/ Modern techniques involving molecular biology, such as metagenomics, have the advantage of exploiting a higher number of microorganisms; however, classic isolation and culture methods used to obtain antimicrobials continue to be promising, especially in the isolation of Actinobacteria, which are responsible for the production of many of these compounds. In this work, two methodologies were used to search for antimicrobial substances—isolation of Actinobacteria and metagenomics of the Atlantic Rainforest soil and of the cultivation of cocoa intercropped with acai berry in the Atlantic Rainforest. The metagenomic libraries were constructed with the CopyControl Fosmid Library kit EPICENTRE, resulting in a total of 2688 clones, 1344 of each soil sample. None of the clones presented antimicrobial activity against the microorganisms tested: S. aureus, Bacillus subtilis, and Salmonella choleraesuis. A total of 46 isolates were obtained from the isolation of soil Actinobacteria: 24 isolates from Atlantic Rainforest soil and 22 isolates from the intercrop cultivation soil. Of these, two Atlantic Rainforest soil isolates inhibited the growth of S. aureus including a clinical isolate of S. aureus MRSA—a promising result, since it is an important multidrug-resistant human pathogen. Danyelle Alves Martins Assis, Rachel Passos Rezende, and João Carlos Teixeira Dias Copyright © 2014 Danyelle Alves Martins Assis et al. All rights reserved. Detection of Tannery Effluents Induced DNA Damage in Mung Bean by Use of Random Amplified Polymorphic DNA Markers Tue, 11 Mar 2014 00:00:00 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2014/727623/ Common effluent treatment plant (CETP) is employed for treatment of tannery effluent. However, the performance of CETP for reducing the genotoxic substances from the raw effluent is not known. In this study, phytotoxic and genotoxic effects of tannery effluents were investigated in mung bean (Vigna radiata (L.) Wilczek). For this purpose, untreated and treated tannery effluents were collected from CETP Unnao (UP), India. Seeds of mung bean were grown in soil irrigated with various concentrations of tannery effluents (0, 25, 50, 75, and 100%) for 15 days. Inhibition of seed germination was 90% by 25% untreated effluent and 75% treated effluent, compared to the control. Plant growth was inhibited by 51% and 41% when irrigated with untreated and treated effluents at 25% concentration. RAPD technique was used to evaluate the genotoxic effect of tannery effluents (untreated and treated) irrigation on the mung bean. The RAPD profiles obtained showed that both untreated and treated were having genotoxic effects on mung bean plants. This was discernible with appearance/disappearance of bands in the treatments compared with control plants. A total of 87 RAPD bands were obtained using eight primers and 42 (48%) of these showed polymorphism. Irrigating plants with untreated effluent caused 12 new bands to appear and 18 to disappear. Treated effluent caused 8 new bands and the loss of 15 bands. The genetic distances shown on the dendrogram revealed that control plants and those irrigated with treated effluent were clustered in one group (joined at distance of 0.28), whereas those irrigated with untreated effluent were separated in another cluster at larger distance (joined at distance of 0.42). This indicates that treated effluent is less genotoxic than the untreated. Nei’s genetic similarity indices calculated between the treatments and the control plants showed that the control and the plants irrigated with treated tannery effluent had a similarity index of 0.75, the control and plants irrigated with untreated 0.65, and between the treatments 0.68. We conclude that both untreated and treated effluents contain genotoxic substances that caused DNA damage to mung beans. CETP Unnao removes some, but not all, genotoxic substances from tannery effluent. Consequently, use of both untreated and treated wastewater for irrigation poses health hazard to human and the environment. Abhay Raj, Sharad Kumar, Izharul Haq, and Mahadeo Kumar Copyright © 2014 Abhay Raj et al. All rights reserved. Effect of Chitosan on Rhizome Rot Disease of Turmeric Caused by Pythium aphanidermatum Thu, 06 Mar 2014 20:07:47 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2014/305349/ Chitosan was evaluated for its potential to induce antifungal hydrolases in susceptible turmeric plant (Curcuma longa L.). Under field conditions, the application of chitosan (crab shell) to turmeric plants by foliar spray method induces defense enzymes such as chitinases and chitosanases. Such an increase in enzyme activity was enhanced by spraying chitosan (0.1% w/v) on leaves of turmeric plants at regular intervals. Gel electrophoresis revealed new chitinase and chitosanase isoforms in leaves of turmeric plants treated with chitosan. Treated turmeric plants showed increased resistance towards rhizome rot disease caused by Pythium aphanidermatum, whereas control plants expressed severe rhizome rot disease. Increased activity of defense enzymes in leaves of chitosan treated turmeric plants may play a role in restricting the development of disease symptoms. The eliciting properties of chitosan make chitosan a potential antifungal agent for the control of rhizome rot disease of turmeric. Sathiyanarayanan Anusuya and Muthukrishnan Sathiyabama Copyright © 2014 Sathiyanarayanan Anusuya and Muthukrishnan Sathiyabama. All rights reserved. Improvement of Polyunsaturated Fatty Acid Production in Echium acanthocarpum Transformed Hairy Root Cultures by Application of Different Abiotic Stress Conditions Wed, 13 Nov 2013 18:22:06 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/169510/ Fatty acids are of great nutritional, therapeutic, and physiological importance, especially the polyunsaturated n-3 fatty acids, possessing larger carbon chains and abundant double bonds or their immediate precursors. A few higher plant species are able to accumulate these compounds, like those belonging to the Echium genus. Here, the novel E. acanthocarpum hairy root system, which is able to accumulate many fatty acids, including stearidonic and α-linolenic acids, was optimized for a better production. The application of abiotic stress resulted in larger yields of stearidonic and α-linolenic acids, 60 and 35%, respectively, with a decrease in linoleic acid, when grown in a nutrient medium consisting of B5 basal salts, sucrose or glucose, and, more importantly, at a temperature of 15∘C. The application of osmotic stress employing sorbitol showed no positive influence on the fatty acid yields; furthermore, the combination of a lower culture temperature and glucose did not show a cumulative boosting effect on the yield, although this carbon source was similarly attractive. The abiotic stress also influenced the lipid profile of the cultures, significantly increasing the phosphatidylglycerol fraction but not the total lipid neither their biomass, proving the appropriateness of applying various abiotic stress in this culture to achieve larger yields. Rafael Zárate, Elena Cequier-Sánchez, Covadonga Rodríguez, Roberto Dorta-Guerra, Nabil El Jaber-Vazdekis, and Ángel G. Ravelo Copyright © 2013 Rafael Zárate et al. All rights reserved. The Medicinal Timber Canarium patentinervium Miq. (Burseraceae Kunth.) Is an Anti-Inflammatory Bioresource of Dual Inhibitors of Cyclooxygenase (COX) and 5-Lipoxygenase (5-LOX) Tue, 01 Oct 2013 10:55:24 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/986361/ The barks and leaves extracts of Canarium patentinervium Miq. (Burseraceae Kunth.) were investigated for cyclooxygenase (COX) and 5-lipoxygenase (LOX) inhibition via in vitro models. The corresponding antioxidative power of the plant extract was also tested via nonenzyme and enzyme in vitro assays. The ethanolic extract of leaves inhibited the enzymatic activity of 5-LOX, COX-1, and COX-2 with IC50 equal to  μg/mL,  μg/mL, and  μg/mL, respectively, with selective COX-2 activity noted in ethanolic extract of barks with COX-1/COX-2 ratio of 1.22. The ethanol extract of barks confronted oxidation in the ABTS, DPPH, and FRAP assay with EC50 values equal to  μg/mL,  μg/mL, and  μg/mL, respectively, while the ethanol extract of leaves confronted oxidation in β-carotene bleaching assay and superoxide dismutase (SOD) assay with EC50 value of  μg/mL and IC50 value of  μg/mL. The ethanol extract acts as a dual inhibitor of LOX and COX enzymes with potent antioxidant capacity. The clinical significance of these data is quite clear that they support a role for Canarium patentinervium Miq. (Burseraceae Kunth.) as a source of lead compounds in the management of inflammatory diseases. R. Mogana, K. Teng-Jin, and C. Wiart Copyright © 2013 R. Mogana et al. All rights reserved. Encapsulation of Phaseolus lunatus Protein Hydrolysate with Angiotensin-Converting Enzyme Inhibitory Activity Tue, 24 Sep 2013 16:07:40 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/341974/ The objective of recent research has been to seek alternative therapeutic treatments; for this reason, the use of protein hydrolysates from diverse sources has been studied. A way to guarantee that these treatments reach the site of action is through protection with covers, such as microcapsules. Therefore, proteins from the legume Phaseolus lunatus were hydrolyzed and encapsulated with a blend of Delonix regia carboxymethylated gum/sodium alginate (50 : 50 w/w). Hydrolysis release conditions in a simulated gastrointestinal system were obtained using CaCl2 concentrations as the main factor, indicating that lower CaCl2 concentrations lead to an increased hydrolysis release. Beads obtained with 1.0 mM of CaCl2 exhibited a better hydrolysate release rate under intestinal simulated conditions and the proteins maintained an IC50 of 2.9 mg/mL. Capsules obtained with the blend of Delonix regia carboxymethylated gum/sodium alginate would be used for the controlled delivery of hydrolysates with potential use as nutraceutical or therapeutic agents. Jorge Carlos Ruiz Ruiz, Maira Rubí Segura Campos, David Abram Betancur Ancona, and Luis Antonio Chel Guerrero Copyright © 2013 Jorge Carlos Ruiz Ruiz et al. All rights reserved. Use of PCR-DGGE Based Molecular Methods to Analyse Microbial Community Diversity and Stability during the Thermophilic Stages of an ATAD Wastewater Sludge Treatment Process as an Aid to Performance Monitoring Mon, 23 Sep 2013 11:52:46 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/162645/ PCR and PCR-DGGE techniques have been evaluated to monitor biodiversity indexes within an ATAD (autothermal thermophilic aerobic digestion) system treating domestic sludge for land spread, by examining microbial dynamics in response to elevated temperatures during treatment. The ATAD process utilises a thermophilic population to generate heat and operates at elevated pH due to degradation of sludge solids, thus allowing pasteurisation and stabilisation of the sludge. Genera-specific PCR revealed that Archaea, Eukarya and Fungi decline when the temperature reaches 59°C, while the bacterial lineage constitutes the dominant group at this stage. The bacterial community at the thermophilic stage, its similarity index to the feed material, and the species richness present were evaluated by PCR-DGGE. Parameters such as choice of molecular target (16S rDNA or rpoB genes), and electrophoresis condition, were optimised to maximise the resolution of the method for ATAD. Dynamic analysis of microbial communities was best observed utilising PCR-DGGE analysis of the V6-V8 region of 16S rDNA, while rpoB gene profiles were less informative. Unique thermophilic communities were shown to quickly adapt to process changes, and shown to be quite stable during the process. Such techniques may be used as a monitoring technique for process health and efficiency. Anna V. Piterina and J. Tony Pembroke Copyright © 2013 Anna V. Piterina and J. Tony Pembroke. All rights reserved. NaCl Effects on In Vitro Germination and Growth of Some Senegalese Cowpea (Vigna unguiculata (L.) Walp.) Cultivars Thu, 25 Jul 2013 13:42:28 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/382417/ Cowpea (Vigna unguiculata (L.) Walp.) is one of the most important grain legumes in sub-Saharian regions. It contributes to man food security by providing a protein-rich diet. However, its production is limited by abiotic stresses such as salinity. This study aims to evaluate the salt tolerance of 15 cowpea cultivars, at germination stage. The seed germination process consisted of sowing them in agarified water (8 g·L−1) supplemented with 6 different concentrations of NaCl (0, 10, 50, 100, 150, and 200 mM). Results highlighted that high salt concentrations drastically reduced germination and significantly delayed the process for all varieties. A cowpea varietal effect towards the salt tolerance was noticed. Genotypes Diongoma, 58-78, and 58-191 were more salt-tolerant cultivars while Mougne and Yacine were more salt-sensitive ones as confirmed in the three groups of the dendrogram. NaCl effects on the early vegetative growth of seedlings were assessed with a tolerant (58-191) and a susceptible (Yacine) cultivar. Morphological (length and dry biomass) and physiological (chlorophyll and proline contents) parameter measurements revealed a negative effect of high (NaCl). However, 58-191 was much more salt tolerant, and the chlorophyll and proline contents were higher than those of Yacine genotype at increasing salt concentrations. Mahamadou Thiam, Antony Champion, Diaga Diouf, and Mame Ourèye SY Copyright © 2013 Mahamadou Thiam et al. All rights reserved. Decolourisation of Synthetic Dyes by Endophytic Fungal Flora Isolated from Senduduk Plant (Melastoma malabathricum) Mon, 22 Jul 2013 10:24:57 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/260730/ A total of twenty endophytic fungi successfully isolated from Melastoma malabathricum (Senduduk) were examined for their ability to decolourise azo dyes: Congo red, Orange G, and Methyl red and an anthraquinone dye, Remazol Brilliant Blue R. Initial screening on the glucose minimal media agar plates amended with 200 mg L−1 of each respective dye showed that only isolate MS8 was able to decolourise all of the four dyes. The isolate decolourised completely both the RBBR and Orange G in the agar medium within 8 days. Further quantitative analysis of the dye decolourisation by isolate MS8 in aqueous minimal medium showed that isolate MS8 was able to decolourise all the tested dyes at varying levels. Dye decolourisation by the isolate MS8 was determined to be 97% for RBBR, 33% for Orange G, 48% for Congo red, and 56% for Methyl red, respectively, within a period of 16 days. Molecular identification of the fungal isolate MS8 using primer ITS1 and ITS4 showed that isolate MS8 shared 99% sequence similarity with Marasmius cladophyllus, a Basidiomycete. The ability to decolourise different types of dyes by isolate MS8 thus suggested a possible application of this fungus in the decolourisation of dyestuff effluents. Ngui Sing Ngieng, Azham Zulkharnain, Hairul Azman Roslan, and Ahmad Husaini Copyright © 2013 Ngui Sing Ngieng et al. All rights reserved. Potential of Aqueous Ozone to Control Aflatoxigenic Fungi in Brazil Nuts Wed, 17 Jul 2013 09:20:44 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/859830/ This study aimed to verify the use of aqueous ozone as alternative technology for fungal control. Brazil nuts sterilized were inoculated with either or  conidia mL−1 of Aspergillus flavus (MUM 9201) to determine optimal treatment parameters and different aqueous ozone contact times. These assays showed that the effect of ozone is almost immediate against A. flavus, and the optimum ozone concentration depended on the number of initial viable spores on the shell. The remaining viable spores in the ozone solution were recorded, and the rate of inactivation for each treatment was determined by assessing the ratio between the cfu of each treatment and the control. The ozonized nuts were also cultured to recover the fungal population. Aqueous ozone was effective in reducing the conidia of A. flavus and the natural fungal population associated with Brazil nuts. Aqueous ozone presented a great potential to reduce microorganisms counts in Brazil nuts with a great potential use in packing houses for decontamination step. Otniel Freitas-Silva, Héctor Morales-Valle, and Armando Venâncio Copyright © 2013 Otniel Freitas-Silva et al. All rights reserved. Differential Inhibition of Helicoverpa armigera (Hubner) Gut Proteinases by Proteinase Inhibitors of Okra and It's Wild Relatives Sun, 17 Mar 2013 16:14:25 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/632173/ The seeds of ten genotypes and twenty-nine wild relatives of okra were analysed for the presence of trypsin, chymotrypsin, and Helicoverpa gut proteinases (HGPs) inhibitors (HGPIs), with the aim to identify potent inhibitors of H. armigera gut proteinases. Proteinase inhibitors (PIs) obtained from wild relatives of okra exhibited stronger inhibition of HGPs than the genotypes of okra. In in vitro inhibitory assay against HGPs, A. tuberculatus 90396 and 90515 showed high tryptic inhibitory (71.8% and 69.2%), chymotryptic inhibitory (68.5% and 66.2%), and Helicoverpa gut proteinase activity (70.2% and 68.2%). In electrophoretic profile showed the same variation in the number of trypsin inhibitors (TIs), chymotrypsin Inhibitors (CIs), and HGPIs isoforms with different intensities, whereas genotypes of okra mostly showed monomorphic profile. Maximum eight HGPIs isoforms were found in A. tuberculatus (90396 and 90515). In bioassay studies, significant reduction in weight of H. armigera larvae was found, when larvae fed on PIs obtained from A. tuberculatus (90396 and 90515). Thus, the result of the present investigation indicates that further exploration of PIs obtained from A. tuberculatus (90396 and 90515) will be helpful for developing PIs-based insect resistance management strategies. Shilpa K. Udamale, M. P. Moharil, T. B. Ugale, and J. M. Mankar Copyright © 2013 Shilpa K. Udamale et al. All rights reserved. Zinc Uptake by Lactic Acid Bacteria Wed, 13 Mar 2013 11:34:22 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/312917/ The study aims to investigate zinc biosorption by strains of lactobacilli and bifidobacteria with a view to exploit them as organic matrixes for zinc dietary supplementation. Sixteen human strains of Lactobacillus and Bifidobacterium were assayed for zinc uptake. The minimum inhibitory concentration of zinc salts differed among the strains, but was never below 15 mmol L−1. When cultured in MRS broth containing 10 mmol L−1 ZnSO4, all the strains were capable of accumulating zinc in the range between 11 and 135 μmol g−1. The highest amount of cell-bound zinc was obtained in L. acidophilus WC 0203. pH-controlled batch cultures of this strain revealed that zinc uptake started in the growth phase, but occurred mostly during the stationary phase. Pasteurized and viable cultures accumulated similar amount of zinc, suggesting that a nonmetabolically mediated mechanism is involved in zinc uptake. These results provide new perspectives on the specific use of probiotics, since L. acidophilus WC 0203 could function as an organic matrix for zinc incorporation. The bioavailability of Lactobacillus-bound zinc deserves to be investigated to provide a future basis for optimization of zinc supplementation or fortification. Alan Leonardi, Simona Zanoni, Marzia De Lucia, Alberto Amaretti, Stefano Raimondi, and Maddalena Rossi Copyright © 2013 Alan Leonardi et al. All rights reserved. Genetically Modified Plants: Public and Scientific Perceptions Thu, 07 Mar 2013 10:05:16 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/820671/ The potential of genetically modified plants to meet the requirements of growing population is not being recognized at present. This is a consequence of concerns raised by the public and the critics about their applications and release into the environment. These include effect on human health and environment, biosafety, world trade monopolies, trustworthiness of public institutions, integrity of regulatory agencies, loss of individual choice, and ethics as well as skepticism about the real potential of the genetically modified plants, and so on. Such concerns are enormous and prevalent even today. However, it should be acknowledged that most of them are not specific for genetically modified plants, and the public should not forget that the conventionally bred plants consumed by them are also associated with similar risks where no information about the gene(s) transfer is available. Moreover, most of the concerns are hypothetical and lack scientific background. Though a few concerns are still to be disproved, it is viewed that, with proper management, these genetically modified plants have immense potential for the betterment of mankind. In the present paper, an overview of the raised concerns and wherever possible reasons assigned to explain their intensity or unsuitability are reviewed. Smita Rastogi Verma Copyright © 2013 Smita Rastogi Verma. All rights reserved. Biofilm-Mediated Enhanced Crude Oil Degradation by Newly Isolated Pseudomonas Species Tue, 05 Mar 2013 14:43:07 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/250749/ The bioavailability of organic contaminants to the degrading bacteria is a major limitation to efficient bioremediation of sites contaminated with hydrophobic pollutants. Such limitation of bioavailability can be overcome by steady-state biofilm-based reactor. The aim of this study was to examine the effect of such multicellular aggregation by naturally existing oil-degrading bacteria on crude oil degradation. Microorganisms, capable of utilizing crude oil as sole carbon source, were isolated from river, estuary and sea-water samples. Biochemical and 16S rDNA analysis of the best degraders of the three sources was found to belong to the Pseudomonas species. Interestingly, one of the isolates was found to be close to Pseudomonas otitidis family which is not reported yet as a degrader of crude oil. Biodegradation of crude oil was estimated by gas chromatography, and biofilm formation near oil-water interface was quantified by confocal laser scanning microscopy. Biofilm supported batches of the isolated Pseudomonas species were able to degrade crude oil much readily and extensively than the planktonic counterparts. Volumetric and topographic analysis revealed that biofilms formed in presence of crude oil accumulate higher biomass with greater thickness compared to the biofilms produced in presence of glucose as sole carbon source. Debdeep Dasgupta, Ritabrata Ghosh, and Tapas K. Sengupta Copyright © 2013 Debdeep Dasgupta et al. All rights reserved. Enhanced Cellulase Production from Bacillus subtilis by Optimizing Physical Parameters for Bioethanol Production Thu, 21 Feb 2013 14:48:59 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/965310/ Effect of physical parameters such as initial pH, agitation (rpm), and temperature (°C) for cellulase production from Bacillus subtilis AS3 was investigated. Central composite design of experiments followed by multiple desirability function was applied for the optimization of cellulase activity and cell growth. The effect of the temperature and agitation was found to be significant among the three independent variables. The optimum levels of initial pH, temperature, and agitation for alkaline carboxymethylcellulase (CMCase) production predicted by the model were 7.2, 39°C, and 121 rpm, respectively. The CMCase activity with unoptimized physical parameters and previously optimized medium composition was 0.43 U/mL. The maximum activity (0.56 U/mL) and cell growth (2.01 mg/mL) predicted by the model were in consensus with values (0.57 U/mL, 2.1 mg/mL) obtained using optimized medium and optimal values of physical parameters. After optimization, 33% enhancement in CMCase activity (0.57 U/mL) was recorded. On scale-up of cellulase production process in bioreactor with all the optimized conditions, an activity of 0.75 U/mL was achieved. Consequently, the bacterial cellulase employed for bioethanol production expending (5%, w/v) NaOH-pretreated wild grass with Zymomonas mobilis yielded an utmost ethanol titre of 7.56 g/L and 11.65 g/L at shake flask and bioreactor level, respectively. Deepmoni Deka, Saprativ P. Das, Naresh Sahoo, Debasish Das, Mohammad Jawed, Dinesh Goyal, and Arun Goyal Copyright © 2013 Deepmoni Deka et al. All rights reserved. Fermentation and Hydrogen Metabolism Affect Uranium Reduction by Clostridia Thu, 21 Feb 2013 07:59:41 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/657160/ Previously, it has been shown that not only is uranium reduction under fermentation condition common among clostridia species, but also the strains differed in the extent of their capability and the pH of the culture significantly affected uranium(VI) reduction. In this study, using HPLC and GC techniques, metabolic properties of those clostridial strains active in uranium reduction under fermentation conditions have been characterized and their effects on capability variance of uranium reduction discussed. Then, the relationship between hydrogen metabolism and uranium reduction has been further explored and the important role played by hydrogenase in uranium(VI) and iron(III) reduction by clostridia demonstrated. When hydrogen was provided as the headspace gas, uranium(VI) reduction occurred in the presence of whole cells of clostridia. This is in contrast to that of nitrogen as the headspace gas. Without clostridia cells, hydrogen alone could not result in uranium(VI) reduction. In alignment with this observation, it was also found that either copper(II) addition or iron depletion in the medium could compromise uranium reduction by clostridia. In the end, a comprehensive model was proposed to explain uranium reduction by clostridia and its relationship to the overall metabolism especially hydrogen (H2) production. Weimin Gao and Arokiasamy J. Francis Copyright © 2013 Weimin Gao and Arokiasamy J. Francis. All rights reserved. Optimization of Cellulase Production from Bacteria Isolated from Soil Tue, 19 Feb 2013 15:20:31 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/985685/ Cellulase-producing bacteria were isolated from soil and identified as Pseudomonas fluorescens, Bacillus subtilIs, E. coli, and Serratia marcescens. Optimization of the fermentation medium for maximum cellulase production was carried out. The culture conditions like pH, temperature, carbon sources, and nitrogen sources were optimized. The optimum conditions found for cellulase production were 40°C at pH 10 with glucose as carbon source and ammonium sulphate as nitrogen source, and coconut cake stimulates the production of cellulase. Among bacteria, Pseudomonas fluorescens is the best cellulase producer among the four followed by Bacillus subtilis, E. coli, and Serratia marscens. Sonia Sethi, Aparna Datta, B. Lal Gupta, and Saksham Gupta Copyright © 2013 Sonia Sethi et al. All rights reserved. Application of Plackett-Burman Experimental Design for Lipase Production by Aspergillus niger Using Shea Butter Cake Wed, 13 Feb 2013 16:01:46 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/718352/ Plackett-Burman design was used to efficiently select important medium components affecting the lipase production by Aspergillus niger using shea butter cake as the main substrate. Out of the eleven medium components screened, six comprising of sucrose, (NH4)2SO4, Na2HPO4, MgSO4, Tween-80, and olive oil were found to contribute positively to the overall lipase production with a maximum production of 3.35 U/g. Influence of tween-80 on lipase production was investigated, and 1.0% (v/w) of tween-80 resulted in maximum lipase production of 6.10 U/g. Thus, the statistical approach employed in this study allows for rapid identification of important medium parameters affecting the lipase production, and further statistical optimization of medium and process parameters can be explored using response surface methodology. Aliyu Salihu, Muntari Bala, and Shuaibu M. Bala Copyright © 2013 Aliyu Salihu et al. All rights reserved. Thermostable Alkaline Phytase from Alcaligenes sp. in Improving Bioavailability of Phosphorus in Animal Feed: In Vitro Analysis Wed, 13 Feb 2013 15:53:01 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/394305/ A bacterial isolate, Alcaligenes sp. secreting phytase (EC 3.1.3.8), was isolated and characterized. The optimum conditions for the production of phytase included a fermentation period of 96 h, pH 8.0, and the addition of 1% (w/v) maltose and 1% (w/v) beef extract to the culture medium. This enzyme was purified to homogeneity and had an apparent molecular mass of 41 kDa. The optimum pH range and temperature for the activity of phytase were found to be 7.0-8.0 and 60°C, respectively. This enzyme was strongly inhibited by 0.005 M of Mn2+, Mg2+, and Zn2+. In vitro studies revealed that the phytase from Alcaligenes sp. released inorganic phosphate from plant phytates. Phytase released 1930 ± 28, 1740 ± 13, 1050 ± 31, 845 ± 7, 1935 ± 32, and 1655 ± 21 mg inorganic phosphate/kg plant phytates, namely, chick pea, corn, green pea, groundnut, pearl pea, and chick feed, respectively. Ponnuswamy Vijayaraghavan, R. Raja Primiya, and Samuel Gnana Prakash Vincent Copyright © 2013 Ponnuswamy Vijayaraghavan et al. All rights reserved. Sequentially Integrated Optimization of the Conditions to Obtain a High-Protein and Low-Antinutritional Factors Protein Isolate from Edible Jatropha curcas Seed Cake Wed, 13 Feb 2013 09:39:50 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/197201/ Jatropha curcas seed cake is a protein-rich byproduct of oil extraction which could be used to produce protein isolates. The purpose of this study was the optimization of the protein isolation process from the seed cake of an edible provenance of J. curcas by an alkaline extraction followed by isoelectric precipitation method via a sequentially integrated optimization approach. The influence of four different factors (solubilization pH, extraction temperature, NaCl addition, and precipitation pH) on the protein and antinutritional compounds content of the isolate was evaluated. The estimated optimal conditions were an extraction temperature of 20°C, a precipitation pH of 4, and an amount of NaCl in the extraction solution of 0.6 M for a predicted protein content of 93.3%. Under these conditions, it was possible to obtain experimentally a protein isolate with 93.21% of proteins, 316.5 mg 100 g−1 of total phenolics, 2891.84 mg 100 g−1 of phytates and 168 mg 100 g−1 of saponins. The protein content of the this isolate was higher than the content reported by other authors. Liliana León-López, Gloria Dávila-Ortiz, Cristian Jiménez-Martínez, and Humberto Hernández-Sánchez Copyright © 2013 Liliana León-López et al. All rights reserved. Identification of Appropriate Reference Genes for qRT-PCR Analysis of Heat-Stressed Mammary Epithelial Cells in Riverine Buffaloes (Bubalus bubalis) Mon, 28 Jan 2013 09:25:52 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/735053/ Gene expression studies require appropriate normalization methods for proper evaluation of reference genes. To date, not many studies have been reported on the identification of suitable reference genes in buffaloes. The present study was undertaken to determine the panel of suitable reference genes in heat-stressed buffalo mammary epithelial cells (MECs). Briefly, MEC culture from buffalo mammary gland was exposed to 42 °C for one hour and subsequently allowed to recover at 37 °C for different time intervals (from 30 m to 48 h). Three different algorithms, geNorm, NormFinder, and BestKeeper softwares, were used to evaluate the stability of 16 potential reference genes from different functional classes. Our data identified RPL4, EEF1A1, and RPS23 genes to be the most appropriate reference genes that could be utilized for normalization of qPCR data in heat-stressed buffalo MECs. Neha Kapila, Amit Kishore, Monika Sodhi, Ankita Sharma, Pawan Kumar, A. K. Mohanty, Tanushri Jerath, and M. Mukesh Copyright © 2013 Neha Kapila et al. All rights reserved. Yield and Properties of Ethanol Biofuel Produced from Different Whole Cassava Flours Mon, 21 Jan 2013 12:10:05 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/916481/ The yield and properties of ethanol biofuel produced from five different whole cassava flours were investigated. Ethanol was produced from five different whole cassava flours. The effect of quantity of yeast on ethanol yield, effect of whole cassava flour to acid and mineralized media ratio on the yield of ethanol produced, and the physical properties of ethanol produced from different cassava were investigated. Physical properties such as distillation range, density, viscosity, and flash point of ethanol produced differ slightly for different cultivars, while the yield of ethanol and electrical conductivity of ethanol from the different cassava cultivars varies significantly. The variation in mineral composition of the different whole cassava flours could also lead to variation in the electrical conductivity of ethanol produced from the different cassava cultivars. The differences in ethanol yield are attributed to differences in starch content, protein content, and dry matter of cassava cultivars. High yield of ethanol from whole cassava flour is best produced from cultivars with high starch content, low protein content, and low fiber. F. T. Ademiluyi and H. D. Mepba Copyright © 2013 F. T. Ademiluyi and H. D. Mepba. All rights reserved. Variations in the Regulatory Region of Alpha S1-Casein Milk Protein Gene among Tropically Adapted Indian Native (Bos Indicus) Cattle Mon, 14 Jan 2013 14:57:44 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/926025/ Regulatory region of milk protein alpha S1-casein (αS1-CN) gene was sequenced, characterized, and analyzed to detect variations among 13 Indian cattle (Bos indicus) breeds. Comparative analysis of 1,587 bp region comprising promoter (1,418 bp), exon-I (53 bp), and partial intron-I (116 bp) revealed 35 nucleotide substitutions (32 within promoter region, 1 in exon-I, and 2 in partial intron-I region) and 4 Indels. Within promoter, 15 variations at positions −1399 (A > G), −1288 (G > A), −1259 (T > C), −1158 (T > C), −1016 (A > T), −941 (T > G), −778 (C > T), −610 (G > A), −536 (A > G), −521 (A > G), −330 (A > C), −214 (A > G), −205 (A > T), −206 (C > A), and −175 (A > G) were located within the potential transcription factor binding sites (TFBSs), namely, NF-κE1/c-Myc, GATA-1, GATA-1/NF-E, Oct-1/POU3F2, MEF-2/YY1, GATA-1, AP-1, POU1F1a/GR, TMF, GAL4, YY1/Oct-1, HNF-1, GRalpha/AR, GRalpha/AR, and AP-1, respectively. Seventy-four percent (26/35) of the observed SNPs were novel to Indian cattle and 11 of these novel SNPs were located within one or more TFBSs. Collectively, these might influence the binding affinity towards their respective nuclear TFs thus modulating the level of transcripts in milk and affecting overall protein composition. The study provides information on several distinct variations across indicine and taurine αS1-CN regulatory domains. Amit Kishore, Manishi Mukesh, Ranbir C. Sobti, Bishnu P. Mishra, and Monika Sodhi Copyright © 2013 Amit Kishore et al. All rights reserved. Challenges in Enzymatic Route of Mannitol Production Sun, 30 Dec 2012 15:54:56 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/914187/ Mannitol is an important biochemical often used as medicine and in food sector, yet its biotechnological is not preffered in Industry for large scale production, which may be due to the multistep mechanism involved in hydrogenation and reduction. This paper is a comparative preview covering present chemical and biotechnological approaches existing today for mannitol production at industrial scale. Biotechnological routes are suitable for adaptation at industrial level for mannitol production, and whatever concerns are there had been discussed in detail, namely, raw materials, broad range of enzymes with high activity at elevated temperature suitable for use in reactor, cofactor limitation, reduced by-product formation, end product inhibition, and reduced utilization of mannitol for enhancing the yield with maximum volumetric productivity. Sheelendra Mangal Bhatt, Anand Mohan, and Suresh Kumar Srivastava Copyright © 2013 Sheelendra Mangal Bhatt et al. All rights reserved. Linen Most Useful: Perspectives on Structure, Chemistry, and Enzymes for Retting Flax Sun, 30 Dec 2012 13:45:02 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/186534/ The components of flax (Linum usitatissimum) stems are described and illustrated, with reference to the anatomy and chemical makeup and to applications in processing and products. Bast fiber, which is a major economic product of flax along with linseed and linseed oil, is described with particular reference to its application in textiles, composites, and specialty papers. A short history of retting methods, which is the separation of bast fiber from nonfiber components, is presented with emphasis on water retting, field retting (dew retting), and experimental methods. Past research on enzyme retting, particularly by the use of pectinases as a potential replacement for the current commercial practice of field retting, is reviewed. The importance and mechanism of Ca2+ chelators with pectinases in retting are described. Protocols are provided for retting of both fiber-type and linseed-type flax stems with different types of pectinases. Current and future applications are listed for use of a wide array of enzymes to improve processed fibers and blended yarns. Finally, potential lipid and aromatic coproducts derived from the dust and shive waste streams of fiber processing are indicated. Danny E. Akin Copyright © 2013 Danny E. Akin. All rights reserved. Kinetic Study of Acid Hydrolysis of Rice Straw Sat, 22 Dec 2012 11:45:24 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/170615/ Rice straw is a renewable, cheap, and abundant waste in tropical countries. The pentose content of rice straw can be used as a substrate for many types of value-added products such as xylitol and biofuel. Dilute acid hydrolysis mainly releases pentose from rice straw. The objective of the study was to determine the effect of H2SO4 concentration and reaction time on the xylose production. The variation of the main product xylose with the reaction time was described by a kinetic model and kinetic parameters were calculated to describe the variation of the xylose production with time. The optimum yield (19.35 g/L) was obtained at 0.24 mol/L H2SO4 and 30 minutes. Nibedita Sarkar and Kaustav Aikat Copyright © 2013 Nibedita Sarkar and Kaustav Aikat. All rights reserved. Induction of Defense-Related Enzymes in Banana Plants: Effect of Live and Dead Pathogenic Strain of Fusarium oxysporum f. sp. cubense Wed, 05 Dec 2012 17:06:25 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/601303/ The aim of the present study was to scrutinize the response of banana (Grand Naine variety) plants when interacting with dead or live pathogen, Fusarium oxysporum f.sp. cubense, a causative agent of Panama disease. Response of plants was evaluated in terms of induction of defense-related marker enzyme activity, namely, peroxidase (POX), polyphenol oxidase (PPO), -1,3 glucanase, chitinase, and phenolics. Plant's interaction with live pathogen resulted in early induction of defense to restrain penetration as well as antimicrobial productions. However, pathogen overcame the defense of plant and caused disease. Interaction with dead pathogen resulted in escalating defense response in plants. Later on plants inoculated with dead pathogen showed resistance to even forced inoculation of live pathogen. Results obtained in the present study suggest that dead pathogen was able to mount defense response in plants and provide resistance to Panama disease upon subsequent exposure. Therefore, preparation from dead pathogen could be a potential candidate as a biocontrol agent or plant vaccine to combat Panama disease. Janki N. Thakker, Samiksha Patel, and Pinakin C. Dhandhukia Copyright © 2013 Janki N. Thakker et al. All rights reserved. Macromolecular Crowding Enhances Catalytic Efficiency and Stability of α-Amylase Wed, 28 Nov 2012 09:51:07 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/737805/ In the present study an attempt was made to investigate the macromolecular crowding effect on functional attributes of α-amylase. High concentrations of sugar based cosolvents, (e.g., trehalose, sucrose, sorbitol, and glycerol) were used to mimic the macromolecular crowding environment (of cellular milieu) under in vitro conditions. To assess the effect of macromolecular crowding, the activity and structural properties of the enzyme were evaluated in the presence of different concentrations of the above cosolvents. Based on the results it is suggested that the macromolecular crowding significantly improves the catalytic efficiency of the enzyme with marginal change in the structure. Out of four cosolvents examined, trehalose was found to be the most effective in consistently enhancing thermal stability of the enzyme. Moreover, the relative effectiveness of the above cosolvents was found to be dependent on their concentration used. Jay Kant Yadav Copyright © 2013 Jay Kant Yadav. All rights reserved. Effect of Potassium Ions on Protoplast Generation during Yeast Induction from Mucor circinelloides Tieghem Wed, 28 Nov 2012 09:50:26 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/734612/ Mucor circinelloides aerobically exhibits coenocytic thallic growth habit with straight and circinate sporangiophores which culminate in globose or pyriform columellae enclosed within sporangial walls. It undergoes dimorphic switch with its conversion to multipolar budding yeast-like cells or thallic conidia. This paper confirms the induction of plurality of reproductive structures of the pleomorphic microorganism in minimal medium. Furthermore, construction of pH differentials at inflection points in the biphasic profiles during sporangiospore-yeast transformation indicated the intensity of H+ release from intracellular medium of the growing microorganism in a study conducted with K+ levels (0.0, 0.5, 0.7, 0.9, 1.0,1.10 g/L)-mediated broths. Optimum proton release was at 0.00 and 1.0 g/L K+-supplemented broths, but specific growth rate was least in the latter. It also coincided with a preponderance of neoplastic units, protoplasts, and terminal budding yeast cells. On either side of this K+ level, variation in morphologies, including neoplasts, protoplasts, septate hyphae, thallic, holothallic, and holoblastic conidia, was greater, although olive-green septate hyphae with vesicular conidiogenous apparatus occurred at all K+ levels tested. This study suggested that following the establishment of transmembrane pH gradient across protoplast membrane, operation of Mitchellian proton pump was further promoted, thus leading to active transport mechanism, a prelude to yeast morphology induction. C. O. Omoifo Copyright © 2013 C. O. Omoifo. All rights reserved. Development of Biological Oxygen Demand Biosensor for Monitoring the Fermentation Industry Effluent Tue, 27 Nov 2012 09:26:07 +0000 http://www.hindawi.com/journals/isrn.biotechnology/2013/236062/ A biosensor was developed for the determination of BOD value of fermentation industry effluent. The developed biosensor was fabricated by immobilizing the microbial consortium on cellulose acetate (CA) membrane in close proximity to a DO probe electrode. The microbial consortium was harvested from the fermentation industry effluent. The BOD biosensor was calibrated by using a solution containing the equivalent amount of glucose/glutamic acid (GGA) as a standard sample solution. The response time was optimized by immobilizing different concentrations of cell biomass on CA membrane. Once the response time was optimized, it was used for determination of BOD of fermentation industry effluent. For analysis of fermentation industry effluent, the response time was observed 7 minutes with detection limit 1 mg/L. Good linear range with GGA standard solution was observed, 0.99 with relative standard deviation (RSD) <%. The observed BOD value by biosensor showed a good comparison with the conventional method for the determination of BOD. Neelam Verma and Ashish Kumar Singh Copyright © 2013 Neelam Verma and Ashish Kumar Singh. All rights reserved.