http://www.hindawi.com The latest articles from Hindawi Publishing Corporation © 2013 , Hindawi Publishing Corporation . All rights reserved. Sun, 17 Mar 2013 16:14:25 +0000 http://www.hindawi.com/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. Wed, 13 Mar 2013 11:34:22 +0000 http://www.hindawi.com/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. Thu, 07 Mar 2013 10:05:16 +0000 http://www.hindawi.com/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. Tue, 05 Mar 2013 14:43:07 +0000 http://www.hindawi.com/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. Thu, 21 Feb 2013 14:48:59 +0000 http://www.hindawi.com/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. Thu, 21 Feb 2013 07:59:41 +0000 http://www.hindawi.com/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. Tue, 19 Feb 2013 15:20:31 +0000 http://www.hindawi.com/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. Wed, 13 Feb 2013 16:01:46 +0000 http://www.hindawi.com/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. Wed, 13 Feb 2013 15:53:01 +0000 http://www.hindawi.com/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. Wed, 13 Feb 2013 09:39:50 +0000 http://www.hindawi.com/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. Mon, 28 Jan 2013 09:25:52 +0000 http://www.hindawi.com/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. Mon, 21 Jan 2013 12:10:05 +0000 http://www.hindawi.com/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. Mon, 14 Jan 2013 14:57:44 +0000 http://www.hindawi.com/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. Sun, 30 Dec 2012 15:54:56 +0000 http://www.hindawi.com/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. Sun, 30 Dec 2012 13:45:02 +0000 http://www.hindawi.com/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. Sat, 22 Dec 2012 11:45:24 +0000 http://www.hindawi.com/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. Wed, 05 Dec 2012 17:06:25 +0000 http://www.hindawi.com/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. Wed, 28 Nov 2012 09:51:07 +0000 http://www.hindawi.com/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. Wed, 28 Nov 2012 09:50:26 +0000 http://www.hindawi.com/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. Tue, 27 Nov 2012 09:26:07 +0000 http://www.hindawi.com/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. Tue, 27 Nov 2012 09:05:02 +0000 http://www.hindawi.com/isrn/biotechnology/2013/450948/ Flavonoids are exploited as antioxidants, antimicrobial, antithrombogenic, antiviral, and antihypercholesterolemic agents. Normally, conventional extraction techniques like soxhlet or shake flask methods provide low yield of flavonoids with structural loss, and thereby, these techniques may be considered as inefficient. In this regard, an attempt was made to optimize the flavonoid extraction using orthogonal design of experiment and subsequent structural elucidation by high-performance liquid chromatography-diode array detector-electron spray ionization/mass spectrometry (HPLC-DAD-ESI/MS) techniques. The shake flask method of flavonoid extraction was observed to provide a yield of  (mg/g tissue). With the two different solvents, namely, ethanol and ethyl acetate, tried for the extraction optimization of flavonoid, ethanol (80.1 mg/g tissue) has been proved better than ethyl acetate (20.5 mg/g tissue). The optimal conditions of the extraction of flavonoid were found to be 85°C, 3 hours with a material ratio of 1 : 20, 75% ethanol, and 1 cycle of extraction. About seven different phenolics like robinin, quercetin, rutin, sinapoyl-hexoside, dicaffeic acid, and two unknown compounds were identified for the first time in the flowers of T. heyneana. The study has also concluded that L16 orthogonal design of experiment is an effective method for the extraction of flavonoid than the shake flask method. Thiyagarajan Sathishkumar, Ramakrishnan Baskar, Mohan Aravind, Suryanarayanan Tilak, Sri Deepthi, and Vellalore Maruthachalam Bharathikumar Copyright © 2013 Thiyagarajan Sathishkumar et al. All rights reserved. Mon, 26 Nov 2012 15:57:53 +0000 http://www.hindawi.com/isrn/biotechnology/2013/515091/ The development of reliable processes for the synthesis of gold nanoparticles is an important aspect of current nanotechnology research. Recently, reports are published on the extracellular as well as intracellular biosynthesis of gold nanoparticles using microorganisms. However, these methods of synthesis are rather slow. In present study, rapid and extracellular synthesis of gold nanoparticles using a plant pathogenic fungus F. oxysporum f. sp. cubense JT1 (FocJT1) is reported. Incubation of FocJT1 mycelium with auric chloride solution produces gold nanoparticles in 60 min. Gold nanoparticles were characterized by UV-Vis spectroscopy, FTIR, and particle size analysis. The particles synthesized were of 22 nm sized, capped by proteins, and posed antimicrobial activity against Pseudomonas sp. Janki N. Thakker, Pranay Dalwadi, and Pinakin C. Dhandhukia Copyright © 2013 Janki N. Thakker et al. All rights reserved. Tue, 23 Oct 2012 09:59:56 +0000 http://www.hindawi.com/isrn/biotechnology/2013/521067/ Flaxseed (Linum usitatissimum) meal, the main byproduct of the flaxseed oil extraction process, is composed mainly of proteins, mucilage, and phenolic compounds. The extraction methods of phenolics either commonly employed the use of mixed solvents (dioxane/ethanol, water/acetone, water/methanol, and water/ethanol) or are done with the aid of alkaline, acid, or enzymatic hydrolysis. This work aimed at the study of optimal conditions for a clean process, using renewable solvents and enzymes, for the extraction of phenolics and proteins from flaxseed meal. After a screening of the most promising commercial preparations based on different carbohydrases and proteases, a central composite rotatable design and a mixture design were applied, achieving as optimal results a solution containing 6.6 and 152 g of phenolics and proteins, respectively. The statistical approach used in the present study for the enzyme-enhanced extraction of phenolics and proteins from the major flaxseed byproduct was effective. By means of the sequential experimental design methodology, the extraction of such compounds was increased 10-fold and 14-fold, when compared to a conventional nonenzymatic extraction. Bernardo Dias Ribeiro, Daniel Weingart Barreto, and Maria Alice Zarur Coelho Copyright © 2013 Bernardo Dias Ribeiro et al. All rights reserved. Mon, 22 Oct 2012 10:32:26 +0000 http://www.hindawi.com/isrn/biotechnology/2013/528708/ The optimization of extracellular lipase production by Fusarium isolani strain SKWF7 isolated from dairy wastewater was carried out in this study. Initially, the physicochemical factors significantly influencing enzyme production were studied by varying one-factor-at-a-time (OFAT). A mesophilic temperature of 40°C, alkaline pH of 8, and incubation period of 72 hours were found to be the optimal conditions for lipase production. Among the media components, the disaccharide sucrose acted as the best carbon source; palm oil as the best inducing lipid substrate; casein and (NH4)2SO4 as the best organic and inorganic nitrogen sources; Ca2+ ion as the best trace element. In the next phase of work, statistical optimization of medium components was performed by employing the Box-Behnken design of Response Surface Methodology (RSM). The optimum concentrations of three significant factors, namely, palm oil, (NH4)2SO4, and CaCO3 were determined by this method to be 5% (v/v), 5.5 g/L, and 0.1 g/L, respectively. RSM-guided design of experiments resulted in a maximum lipase production of 73.3 U/ml, which is a 1.7-fold increase in comparison with that obtained in the unoptimized medium. These results point towards the success of the model in developing a process for the production of lipase, an enzyme of enormous industrial significance. P. Kanmani, S. Karthik, J. Aravind, and K. Kumaresan Copyright © 2013 P. Kanmani et al. All rights reserved. Sun, 21 Oct 2012 07:50:01 +0000 http://www.hindawi.com/isrn/biotechnology/2013/958945/ A high survival rate for transformed papaya plants when transferred to the field is useful in the quest for improving the commercial quality traits. We report in this paper an improved rooting method for the production of transformed Malaysian Eksotika papaya with high survival rate when transferred to the field. Shoots were regenerated from embryogenic calli transformed with antisense and RNAi constructs of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) genes using the Agrobacterium tumefaciens-mediated transformation method. Regenerated transformed shoots, each measuring approximately 3-4 cm in height, were cultured in liquid half-strength Murashige and Skoog (MS) medium or sterile distilled water, and with either perlite or vermiculite supplementation. All the culturing processes were conducted either under sterile or nonsterile condition. The results showed that rooting under sterile condition was better. Shoots cultured in half-strength MS medium supplemented with vermiculite exhibited a 92.5% rooting efficiency while perlite showed 77.5%. The survival rate of the vermiculite-grown transformed papaya plantlets after transfer into soil, contained in polybags, was 94%, and the rate after transfer into the ground was 92%. Morpho-histological analyses revealed that the tap roots were more compact, which might have contributed to the high survival rates of the plantlets. Rogayah Sekeli, Janna Ong Abdullah, Parameswari Namasivayam, Pauziah Muda, and Umi Kalsom Abu Bakar Copyright © 2013 Rogayah Sekeli et al. All rights reserved. Sun, 21 Oct 2012 07:48:53 +0000 http://www.hindawi.com/isrn/biotechnology/2013/453910/ A convenient and accurate reversed-phase high-performance liquid chromatography (RP-HPLC) method for angiotensin I-converting enzyme inhibitory peptides assay was described in this paper. The mobile phase consisted of 70% A (0.05% TFA and 0.05% triethylamine in water, pH = 2.9–3.3) and 30% B (100% acetonitrile) using an Isogradient program. The flow rate was 0.5 mL/min. The absorb wavelength was 226.5 nm; the column temperature was controlled at 25°C. This method for angiotensin I-converting enzyme inhibitory peptides assay was convenient for the Iso-gradient program. The accuracy of the RT-HPLC method was verified by analyzing ACE inhibitory activity of the hydrolysate peptides of silkworm pupae protein, and the results showed that the RT-HPLC method was available for exploring new source of angiotensin I-converting enzyme inhibitory peptides rapidly and veraciously. Wei Wang, Nan Wang, Yu Zhang, Zheng Cai, Qihe Chen, and Guoqing He Copyright © 2013 Wei Wang et al. All rights reserved. Thu, 27 Sep 2012 10:58:46 +0000 http://www.hindawi.com/isrn/biotechnology/2013/980562/ A medium using fish waste as substrate was designed for production of lipase by Enterococcus faecium MTCC 5695 and Pediococcus acidilactici MTCC 11361. Medium components and culture conditions (fish waste protein hydrolysate (FWPH) concentration, fish waste oil (FWO) concentration, pH, temperature, and fermentation time) which affect lipase production were screened using factorial (5 factors ∗ 2 levels) design of which FWPH concentration, FWO concentration, and fermentation time showed significance (). The levels of these factors were optimized further by Box-Behnken design using response surface methodology (RSM). Optimized conditions were found to be 5% v/v FWO, 0.15 mg/mL FWPH and 24 h of fermentation time for MTCC 5695, and 4% v/v FWO, 0.15 mg/mL FWPH and 24 h of fermentation for MTCC 11361, which were further validated. Under optimized conditions, MTCC 5695 and MTCC 11361 showed 3.15- (543.63 to 1715 U/mL) and 2.3- (214.74 to 493 U/mL) fold increase in lipase production, respectively, as compared to unoptimized conditions. Vrinda Ramakrishnan, Louella Concepta Goveas, Bhaskar Narayan, and Prakash M. Halami Copyright © 2013 Vrinda Ramakrishnan et al. All rights reserved. Tue, 18 Sep 2012 14:11:15 +0000 http://www.hindawi.com/isrn/biotechnology/2013/968241/ Bisphenol A (BPA) and triclosan (TCS) are known or suspected potential endocrine disrupting chemicals (EDCs) which may pose a risk to human health and have an environmental impact. Enzyme preparations containing mainly laccases, obtained from Ganoderma stipitatum and Lentinus swartzii, two autochthonous Colombian forest white rot fungi (WRF), previously identified as high enzyme producers, were used to remove BPA and TCS from aqueous solutions. A Box-Behnken factorial design showed that pH, temperature, and duration of treatment were significant model terms for the elimination of BPA and TCS. Our results demonstrated that these EDCs were extensively removed from 5 mg L−1 solutions after a contact time of 6 hours. Ninety-four percent of TCS and 97.8% of BPA were removed with the enzyme solution from G. stipitatum; 83.2% of TCS and 88.2% of BPA were removed with the L. swartzii enzyme solution. After a 6-hour treatment with enzymes from G. stipitatum and L. swartzii, up to 90% of the estrogenic activity of BPA was lost, as shown by the yeast estrogen screen assay. 2,2-Azino-bis-(3-ethylthiazoline-6-sulfonate) (ABTS) was used as a mediator (laccase/mediator system) and significantly improved the laccase catalyzed elimination of BPA and TCS. The elimination of BPA in the absence of a mediator resulted in production of oligomers of molecular weights of 454, 680, and 906 amu as determined by mass spectra analysis. The elimination of TCS in the same conditions produced dimers, trimers, and tetramers of molecular weights of 574, 859, and 1146 amu. Ecotoxicological studies using Daphnia pulex to determine lethal concentration (LC50) showed an important reduction of the toxicity of BPA and TCS solutions after enzymatic treatments. Use of laccases emerges thus as a key alternative in the development of innovative wastewater treatment technologies. Moreover, the exploitation of local biodiversity appears as a potentially promising approach for identifying new efficient strains for biotechnological applications. Carolina Arboleda, H. Cabana, E. De Pril, J. Peter Jones, G. A. Jiménez, A. I. Mejía, S. N. Agathos, and M. J. Penninckx Copyright © 2013 Carolina Arboleda et al. All rights reserved. Thu, 13 Sep 2012 15:57:37 +0000 http://www.hindawi.com/isrn/biotechnology/2013/590587/ E. coli is the most frequently used host for production of enzymes and other proteins by recombinant DNA technology. E. coli is preferable for its relative simplicity, inexpensive and fast high-density cultivation, well-known genetics, and large number of compatible molecular tools available. Despite all these advantages, expression and production of recombinant enzymes are not always successful and often result in insoluble and nonfunctional proteins. There are many factors that affect the success of cloning, expression, and mass production of enzymes by recombinant E. coli. In this paper, these critical factors and approaches to overcome these obstacles are summarized focusing controlled expression of target protein/enzyme in an unmodified form at industrial level. Md. Fakruddin, Reaz Mohammad Mazumdar, Khanjada Shahnewaj Bin Mannan, Abhijit Chowdhury, and Md. Nur Hossain Copyright © 2013 Md. Fakruddin et al. All rights reserved. Wed, 29 Aug 2012 14:07:20 +0000 http://www.hindawi.com/isrn/biotechnology/2013/652340/ Diversity among biosurfactant producing Bacillus spp. from diverse habitats was studied among 77 isolates. Cluster analysis based on phenotypic characteristics using unweighted pair-group method with arithmetic averages (UPGMAs) method was performed. Bacillus isolates possessing high surface tension activity and five reference strains were subjected to amplified 16S rDNA restriction analysis (ARDRA). A correlation between the phenotypic and genotypic characterization of Bacillus spp. is explored. Most of the oil reservoir isolates showing high surface activity clustered with B. licheniformis and B. subtilis, the hot water spring isolates clustered in two ingroups, while the petroleum contaminated soil isolates were randomly distributed in all the three ingroups. Present work revealed that diversity exists in distribution of Bacillus spp. from thermal and hydrocarbon containing habitats where majority of organisms belonged to B. licheniformis and B. subtilis group. Isolate B. licheniformis TT42 produced biosurfactant which reduced the surface tension of water from 72 mNm−1 to 28 mNm−1, and 0.05 mNm−1 interfacial tension against crude oil at 80∘C. This isolate clustered with B. subtilis and B. licheniformis group on the basis of ARDRA. These findings increase the possibility of exploiting the Bacillus spp. from different habitats and their possible use in oil recovery. Sanket J. Joshi, Harish Suthar, Amit Kumar Yadav, Krushi Hingurao, and Anuradha Nerurkar Copyright © 2013 Sanket J. Joshi et al. All rights reserved.