Biotechnology Research International The latest articles from Hindawi © 2017 , Hindawi Limited . All rights reserved. Generation of Recombinant Antibodies against the beta-(1,6)-Branched beta-(1,3)-D-Glucan Schizophyllan from Immunized Mice via Phage Display Tue, 23 May 2017 00:00:00 +0000 beta-(1,6)-Branched beta-(1,3)-D-glucans like schizophyllan from the basidiomycete Schizophyllum commune excite various immunostimulatory effects and have been clinically tested as adjuvants. Some of the glucans are also applicable in food or petrol industry due to their viscosity and temperature stability in aqueous solution. Antibodies against these glucans could be used as tool for analysis of glucan preparations or for further research of its bioactivity. Therefore, an immune phage display library was constructed from mice immunized with schizophyllan. Three recombinant monoclonal antibodies were isolated from this library by affinity selection (panning) on schizophyllan. The half-maximal effective concentration (EC50) values for those antibodies varied between 16.4 ng mL−1 and 21.3 ng mL−1. The clones showed binding specificity not only for schizophyllan but also for other beta-(1,6)-branched beta-(1,3)-D-glucans of similar macromolecular structure. Denaturation of the secondary structure led to a reduced antibody binding, indicating an epitope requiring the correct conformation of the triple helical structure of the glucans. Jörn Josewski, Sabine Buchmeier, André Frenzel, Philip Tinnefeld, Stefan Dübel, and Udo Rau Copyright © 2017 Jörn Josewski et al. All rights reserved. Optimization of Crude Oil and PAHs Degradation by Stenotrophomonas rhizophila KX082814 Strain through Response Surface Methodology Using Box-Behnken Design Wed, 28 Dec 2016 13:10:47 +0000 The present paper describes the process optimization study for crude oil degradation which is a continuation of our earlier work on hydrocarbon degradation study of the isolate Stenotrophomonas rhizophila (PM-1) with GenBank accession number KX082814. Response Surface Methodology with Box-Behnken Design was used to optimize the process wherein temperature, pH, salinity, and inoculum size (at three levels) were used as independent variables and Total Petroleum Hydrocarbon, Biological Oxygen Demand, and Chemical Oxygen Demand of crude oil and PAHs as dependent variables (response). The statistical analysis, via ANOVA, showed coefficient of determination as 0.7678 with statistically significant value 0.0163 fitting in second-order quadratic regression model for crude oil removal. The predicted optimum parameters, namely, temperature, pH, salinity, and inoculum size, were found to be 32.5°C, 9, 12.5, and 12.5 mL, respectively. At this optimum condition, the observed and predicted PAHs and crude oil removal were found to be 71.82% and 79.53% in validation experiments, respectively. The % TPH results correlate with GC/MS studies, BOD, COD, and TPC. The validation of numerical optimization was done through GC/MS studies and   % removal of crude oil. Praveen Kumar Siddalingappa Virupakshappa, Manjunatha Bukkambudhi Krishnaswamy, Gaurav Mishra, and Mohammed Ameenuddin Mehkri Copyright © 2016 Praveen Kumar Siddalingappa Virupakshappa et al. All rights reserved. Corrigendum to “Proteases from Canavalia ensiformis: Active and Thermostable Enzymes with Potential of Application in Biotechnology” Wed, 12 Oct 2016 09:52:43 +0000 Rayane Natashe Gonçalves, Suellen Duarte Gozzini Barbosa, and Raquel Elisa da Silva-López Copyright © 2016 Rayane Natashe Gonçalves et al. All rights reserved. Influence of Halide Solutions on Collagen Networks: Measurements of Physical Properties by Atomic Force Microscopy Sun, 18 Sep 2016 09:10:48 +0000 The influence of aqueous halide solutions on collagen coatings was tested. The effects on resistance against indentation/penetration on adhesion forces were measured by atomic force microscopy (AFM) and the change of Young’s modulus of the coating was derived. Comparative measurements over time were conducted with halide solutions of various concentrations. Physical properties of the mesh-like coating generally showed large variability. Starting with a compact set of physical properties, data disperse after minutes. A trend of increase in elasticity and permeability was found for all halide solutions. These changes were largest in NaI, displaying a logical trend with ion size. However a correlation with concentration was not measured. Adhesion properties were found to be independent of mechanical properties. The paper also presents practical experience for AFM measurements of soft tissue under liquids, particularly related to data evaluation. The weakening in physical strength found after exposure to halide solutions may be interpreted as widening of the network structure or change in the chemical properties in part of the collagen fibres (swelling). In order to design customized surface coatings at optimized conditions also for medical applications, halide solutions might be used as agents with little impact on the safety of patients. Birgit Spitzer-Sonnleitner, André Kempe, and Maximilian Lackner Copyright © 2016 Birgit Spitzer-Sonnleitner et al. All rights reserved. Effect of Purified Mushroom Tyrosinase on Melanin Content and Melanogenic Protein Expression Tue, 06 Sep 2016 14:13:52 +0000 In mammalian melanocytes, melanosome is a highly specialized organelle where melanin is synthesized. Melanin synthesis is controlled by tyrosinase, the vital enzyme in melanogenic pathway. The present investigation is based on an effect of purified mushroom tyrosinase of Agaricus bisporus on B16F10 melanocytes for the melanin production via blocking pigment cell machinery. Using B16F10 melanocytes showed that the stimulation of melanogenesis by purified tyrosinase is due to increased tyrosinase absorption. Cellular tyrosinase activity and melanin content in B16F10 melanocytes were increased by purified tyrosinase in a dose-dependent manner. Western blot analysis revealed that cellular tyrosinase levels were enhanced after treatment with purified tyrosinase for 48 hours. Furthermore, tyrosinase induced phosphorylation of cyclic adenosine monophosphate (cAMP) response element-binding protein (CREB) in a dose-dependent manner. The purified tyrosinase-mediated increase of tyrosinase activity was significantly attenuated by H89, LY294002, Ro-32-0432, and PD98059, cAMP-dependent protein kinase inhibitors. The results indicate that purified tyrosinase can be used as contestant for the treatment of vitiligous skin conditions. Kamal Uddin Zaidi, Sharique A. Ali, and Ayesha S. Ali Copyright © 2016 Kamal Uddin Zaidi et al. All rights reserved. β-Cyclodextrin Production by Cyclodextrin Glucanotransferase from an Alkaliphile Microbacterium terrae KNR 9 Using Different Starch Substrates Thu, 25 Aug 2016 13:39:06 +0000 Cyclodextrin glucanotransferase (CGTase, EC is an important member of α-amylase family which can degrade the starch and produce cyclodextrins (CDs) as a result of intramolecular transglycosylation (cyclization). β-Cyclodextrin production was carried out using the purified CGTase enzyme from an alkaliphile Microbacterium terrae KNR 9 with different starches in raw as well as gelatinized form. Cyclodextrin production was confirmed using thin layer chromatography. Six different starch substrates, namely, soluble starch, potato starch, sago starch, corn starch, corn flour, and rice flour, were tested for CD production. Raw potato starch granules were found to be the best substrate giving 13.46 gm/L of cyclodextrins after 1 h of incubation at 60°C. Raw sago starch gave 12.96 gm/L of cyclodextrins as the second best substrate. To achieve the maximum cyclodextrin production, statistical optimization using Central Composite Design (CCD) was carried out with three parameters, namely, potato starch concentration, CGTase enzyme concentration, and incubation temperature. Cyclodextrin production of 28.22 (gm/L) was achieved with the optimized parameters suggested by the model which are CGTase 4.8 U/L, starch 150 gm/L, and temperature 55.6°C. The suggested optimized conditions showed about 15% increase in β-cyclodextrin production (28.22 gm/L) at 55.6°C as compared to 24.48 gm/L at 60°C. The degradation of raw potato starch granules by purified CGTase was also confirmed by microscopic observations. Kiransinh N. Rajput, Kamlesh C. Patel, and Ujjval B. Trivedi Copyright © 2016 Kiransinh N. Rajput et al. All rights reserved. Proteases from Canavalia ensiformis: Active and Thermostable Enzymes with Potential of Application in Biotechnology Wed, 17 Aug 2016 16:47:22 +0000 Extracts of leaves, seeds, roots, and stem from a tropical legume, C. ensiformis, were prepared employing buffers and detergent in aqueous solution. Leaf extracts had the highest protein content and the most pronounced peptidase activity with optimal pH in the neutral to alkaline range. All extracts exhibited peaks of activity at various pH values, suggesting the presence of distinctive classes of proteases. N-α-Tosyl-L-arginine methyl ester hydrolysis was maximal at 30°C to 60°C and peptidase activity from all extracts presented very good thermal stability after 24 h incubation at 70°C. C. ensiformis proteases exhibited molecular masses of about 200–57, 40–37, and 20–15 kDa by SDS-PAGE analysis. These enzymes cleaved hemoglobin, bovine serum albumin, casein, and gelatin at different levels. Serine and metalloproteases are the major proteases in C. ensiformis extracts, modulated by divalent cations, stable at 1% of surfactant Triton X-100 and at different concentrations of the reducing agent β-mercaptoethanol. Thus, C. ensiformis expresses a particular set of proteases in distinctive organs with high activity and stability, making this legume an important source of proteases with biotechnological potential. Rayane Natshe Gonçalves, Suellen Duarte Gozzini Barbosa, and Raquel Elisa da Silva-López Copyright © 2016 Rayane Natshe Gonçalves et al. All rights reserved. The Use of Adenovirus Dodecahedron in the Delivery of an Enzymatic Activity in the Cell Sun, 08 May 2016 12:52:45 +0000 Penton-dodecahedron (Pt-Dd) derived from adenovirus type 3 is a symmetric complex of pentameric penton base plus fiber which can be produced in the baculovirus system at a high concentration. The size of Pt-Dd is smaller than the virus, but this virus-like particle (VLP) has the major proteins recognized by specific receptors on the surface of almost all types of cell. In this study, by direct observation with fluorescence microscopy on a fixed and living cell, the intracellular trafficking and localization of Pt-Dd labeled with fluorescence dyes in the cytoplasm of HeLa Tub-GFP showed a rapid internalization characteristic. Subsequently, the linkage of horseradish peroxidase (HRP) with Pt-Dd as the vector demonstrated an efficient system to deliver this enzyme into the cell without interfering its enzymatic activity as shown by biochemical and cellular experiments. These results were supported by additional studies using Bs-Dd or free form of the HRP used as the control. Overall, this study strengthens the potential role of Pt-Dd as an alternative vector for delivering therapeutic agents. Sumarheni, Benoit Gallet, and Pascal Fender Copyright © 2016 Sumarheni et al. All rights reserved. Chimerism Analysis of Cell-Free DNA in Patients Treated with Hematopoietic Stem Cell Transplantation May Predict Early Relapse in Patients with Hematologic Malignancies Tue, 23 Feb 2016 11:07:34 +0000 Background. We studied DNA chimerism in cell-free DNA (cfDNA) in patients treated with HSCT. Methods. Chimerism analysis was performed on CD3+ cells, polymorphonuclear (PMN) cells, and cfDNA using 16 small tandem repeat loci. The resulting labeled PCR-products were size-fractionated and quantified. Results. Analyzing samples from 191 patients treated with HSCT for nonneoplastic hematologic disorders demonstrated that the cfDNA chimerism is comparable to that seen in PMN cells. Analyzing leukemia patients ( = 126) showed that, of 84 patients with 100% donor DNA in PMN, 16 (19%) had evidence of clinical relapse and >10% recipient DNA in the plasma. Additional 16 patients of the 84 (19%) showed >10% recipient DNA in plasma, but without evidence of relapse. Eight patients had mixed chimerism in granulocytes, lymphocytes, and plasma, but three of these patients had >10% recipient DNA in plasma compared to PMN cells and these three patients had clinical evidence of relapse. The remaining 34 patients showed 100% donor DNA in both PMN and lymphocytes, but cfDNA showed various levels of chimerism. Of these patients 14 (41%) showed laboratory or clinical evidence of relapse and all had >10% recipient DNA in cfDNA. Conclusion. Monitoring patients after HSCT using cfDNA might be more reliable than cellular DNA in predicting early relapse. Mahmoud Aljurf, Hala Abalkhail, Amal Alseraihy, Said Y. Mohamed, Mouhab Ayas, Fahad Alsharif, Hazza Alzahrani, Abdullah Al-Jefri, Ghuzayel Aldawsari, Ali Al-Ahmari, Asim F. Belgaumi, Claudia Ulrike Walter, Hassan El-Solh, Walid Rasheed, and Maher Albitar Copyright © 2016 Mahmoud Aljurf et al. All rights reserved. Screening and Selection of Medium Components for Cyclodextrin Glucanotransferase Production by New Alkaliphile Microbacterium terrae KNR 9 Using Plackett-Burman Design Wed, 03 Feb 2016 09:43:14 +0000 Cyclodextrin glucanotransferase (CGTase, EC production using new alkaliphile Microbacterium terrae KNR 9 was investigated by submerged fermentation. Statistical screening for components belonging to different categories, namely, soluble and raw starches as carbon sources, complex organic and inorganic nitrogen sources, minerals, a buffering agent, and a surfactant, has been carried out for CGTase production using Plackett-Burman factorial design. To screen out (19), number of variables, (20), number of experiments, were performed. Among the fourteen components screened, four components, namely, soluble starch, corn flour, yeast extract, and K2HPO4, were identified as significant with reference to their concentration effect and corresponding value. Although soluble starch showed highest significance, comparable significance was also observed with corn flour and hence it was selected as a sole carbon source along with yeast extract and K2HPO4 for further media optimization studies. Using screened components, CGTase production was increased to 45% and 87% at shake flask level and laboratory scale fermenter, respectively, as compared to basal media. Kiransinh N. Rajput, Kamlesh C. Patel, and Ujjval B. Trivedi Copyright © 2016 Kiransinh N. Rajput et al. All rights reserved. Utilization of Crude Glycerol as a Substrate for the Production of Rhamnolipid by Pseudomonas aeruginosa Thu, 28 Jan 2016 11:59:47 +0000 Biosurfactants are produced by bacteria or yeast utilizing different substrates as sugars, glycerol, or oils. They have important applications in the detergent, oil, and pharmaceutical industries. Glycerol is the product of biodiesel industry and the existing glycerol market cannot accommodate the excess amounts generated; consequently, new markets for refined glycerol need to be developed. The aim of present work is to optimize the production of microbial rhamnolipid using waste glycerol. We have developed a process for the production of rhamnolipid biosurfactants using glycerol as the sole carbon source by a local Pseudomonas aeruginosa isolate that was obtained from an extensive screening program. A factorial design was applied with the goal of optimizing the rhamnolipid production. The highest production yield was obtained after 2 days when cells were grown in minimal salt media at pH 6, containing 1% (v/v) glycerol and 2% (w/v) sodium nitrate as nitrogen source, at 37°C and at 180 rpm, and reached 2.164 g/L after 54 hours (0.04 g/L h). Analysis of the produced rhamnolipids by TLC, HPLC, and FTIR confirmed the nature of the biosurfactant as monorhamnolipid. Glycerol can serve as a source for the production of rhamnolipid from microbial isolates providing a cheap and reliable substrate. Walaa A. Eraqi, Aymen S. Yassin, Amal E. Ali, and Magdy A. Amin Copyright © 2016 Walaa A. Eraqi et al. All rights reserved. Synergistic Effects of Plant Growth Promoting Rhizobacteria and Chitosan on In Vitro Seeds Germination, Greenhouse Growth, and Nutrient Uptake of Maize (Zea mays L.) Wed, 20 Jan 2016 09:37:08 +0000 This study aimed to assess the effects of three plant growth promoting rhizobacteria (PGPR) and chitosan either singly or in combination on maize seeds germination and growth and nutrient uptake. Maize seeds were treated with chitosan and bacterial solution. The germination and growth tests were carried out in square Petri dishes and plastic pots. The combination chitosan-A. lipoferum-P. fluorescens has increased the seeds vigor index up to 36.44% compared to the control. In comparison to the control, P. putida has significantly improved root weight (44.84%) and germinated seed weight (31.39%) whereas chitosan-P. putida has increased the shoot weight (65.67%). For the growth test, the maximal heights (17.66%) were obtained by plants treated with the combination A. lipoferum-P. fluorescens-P. putida. Chitosan-P. fluorescens induced the highest increases of leaves per plant (50.09%), aerial (84.66%), and underground biomass (108.77%) production. The plants inoculated with A. lipoferum had the large leaf areas with an increase of 54.08%, while combinations P. fluorescens-P. putida and chitosan-A. lipoferum improved the aerial and underground dry matter of plants to 26.35% and 18.18%. The nitrogen content of the plants was increased by chitosan-A. lipoferum-P. fluorescens-P. putida with an increasing of 41.61%. The combination of chitosan and PGPR can be used as biological fertilizers to increase maize production. Nadège A. Agbodjato, Pacôme A. Noumavo, Adolphe Adjanohoun, Léonce Agbessi, and Lamine Baba-Moussa Copyright © 2016 Nadège A. Agbodjato et al. All rights reserved. Role of Heat-Shock Proteins in Cellular Function and in the Biology of Fungi Thu, 31 Dec 2015 06:22:40 +0000 Stress (biotic or abiotic) is an unfavourable condition for an organism including fungus. To overcome stress, organism expresses heat-shock proteins (Hsps) or chaperons to perform biological function. Hsps are involved in various routine biological processes such as transcription, translation and posttranslational modifications, protein folding, and aggregation and disaggregation of proteins. Thus, it is important to understand holistic role of Hsps in response to stress and other biological conditions in fungi. Hsp104, Hsp70, and Hsp40 are found predominant in replication and Hsp90 is found in transcriptional and posttranscriptional process. Hsp90 and Hsp70 in combination or alone play a major role in morphogenesis and dimorphism. Heat stress in fungi expresses Hsp60, Hsp90, Hsp104, Hsp30, and Hsp10 proteins, whereas expression of Hsp12 protein was observed in response to cold stress. Hsp30, Hsp70, and Hsp90 proteins showed expression in response to pH stress. Osmotic stress is controlled by small heat-shock proteins and Hsp60. Expression of Hsp104 is observed under high pressure conditions. Out of these heat-shock proteins, Hsp90 has been predicted as a potential antifungal target due to its role in morphogenesis. Thus, current review focuses on role of Hsps in fungi during morphogenesis and various stress conditions (temperature, pH, and osmotic pressure) and in antifungal drug tolerance. Shraddha Tiwari, Raman Thakur, and Jata Shankar Copyright © 2015 Shraddha Tiwari et al. All rights reserved. Prospects for Irradiation in Cellulosic Ethanol Production Tue, 29 Dec 2015 13:30:22 +0000 Second generation bioethanol production technology relies on lignocellulosic biomass composed of hemicelluloses, celluloses, and lignin components. Cellulose and hemicellulose are sources of fermentable sugars. But the structural characteristics of lignocelluloses pose hindrance to the conversion of these sugar polysaccharides into ethanol. The process of ethanol production, therefore, involves an expensive and energy intensive step of pretreatment, which reduces the recalcitrance of lignocellulose and makes feedstock more susceptible to saccharification. Various physical, chemical, biological, or combined methods are employed to pretreat lignocelluloses. Irradiation is one of the common and promising physical methods of pretreatment, which involves ultrasonic waves, microwaves, γ-rays, and electron beam. Irradiation is also known to enhance the effect of saccharification. This review explains the role of different radiations in the production of cellulosic ethanol. Anita Saini, Neeraj K. Aggarwal, Anuja Sharma, and Anita Yadav Copyright © 2015 Anita Saini et al. All rights reserved. Response Surface Optimization of Bioethanol Production from Sugarcane Molasses by Pichia veronae Strain HSC-22 Tue, 08 Dec 2015 13:35:42 +0000 Pichia veronae strain HSC-22 (accession number KP012558) showed a good tolerance to relatively high temperature, ethanol and sugar concentrations. Response surface optimization based on central composite design of experiments predicted the optimal values of the influencing parameters that affect the production of bioethanol from sugarcane molasses to be as follows: initial pH 5, 25% (w : v) initial molasses concentration, 35°C, 116 rpm, and 60 h. Under these optimum operating conditions the maximum bioethanol production on a batch fermenter scale was recorded as 32.32 g/L with 44% bioethanol yield. Hamed I. Hamouda, Hussein N. Nassar, Hekmat R. Madian, Salem S. Abu Amr, and Nour Sh. El-Gendy Copyright © 2015 Hamed I. Hamouda et al. All rights reserved. Enhanced Bioremediation of Soil Artificially Contaminated with Petroleum Hydrocarbons after Amendment with Capra aegagrus hircus (Goat) Manure Sun, 06 Dec 2015 09:11:02 +0000 This study was carried out to evaluate the biostimulant potentials of Capra aegagrus hircus manure for bioremediation of crude oil contaminated soil (COCS) under tropical conditions. 1 kg of COCS sample was amended with 0.02 kg of C. a. hircus manure and monitored at 14-day intervals for total petroleum hydrocarbon (TPH), nutrient content, and changes in microbial counts. At the end of the study period, there was 62.08% decrease in the concentration of TPH in the amended sample compared to 8.15% decrease in the unamended sample, with significant differences () in TPH concentrations for both samples at different time intervals. Similarly, there was a gradual decrease in the concentrations of total organic carbon, nitrogen, phosphorus, and potassium in both samples. The culturable hydrocarbon-utilizing bacteria (CHUB) increased steadily from 8.5 × 105 cfu/g to 2.70 × 106 cfu/g and from 8.0 × 105 cfu/g to 1.78 × 106 cfu/g for both samples. Acinetobacter, Achromobacter, Bacillus, Flavobacterium, Klebsiella, Micrococcus, Pseudomonas, and Staphylococcus were isolated from amended sample with Pseudomonas being the predominant isolated bacterial genus. This study demonstrated that C. a. hircus manure is a good biostimulant, which enhanced the activities of indigenous hydrocarbonoclastic bacteria resulting in significant decrease in TPH concentration of COCS. T. P. Nwogu, C. C. Azubuike, and C. J. Ogugbue Copyright © 2015 T. P. Nwogu et al. All rights reserved. Plant Growth Promotion Activity of Keratinolytic Fungi Growing on a Recalcitrant Waste Known as “Hair Waste” Mon, 30 Nov 2015 07:53:15 +0000 Purpureocillium lilacinum (Thom) Samsom is one of the most studied fungi in the control of plant parasitic nematodes. However, there is not specific information on its ability to inhibit some pathogenic bacteria, fungi, or yeast. This work reports the production of several antifungal hydrolytic enzymes by a strain of P. lilacinum when it is grown in a medium containing hair waste. The growth of several plant-pathogenic fungi, Alternaria alternata, Aspergillus niger, and Fusarium culmorum, was considerably affected by the presence of P. lilacinum’s supernatant. Besides antifungal activity, P. lilacinum demonstrates the capability to produce indoleacetic acid and ammonia during time cultivation on hair waste medium. Plant growth-promoting activity by cell-free supernatant was evidenced through the increase of the percentage of tomato seed germination from 71 to 85% after 48 hours. A 21-day plant growth assay using tomato plants indicates that crude supernatant promotes the growth of the plants similar to a reference fertilizer (). These results suggest that both strain and the supernatant may have potential to be considered as a potent biocontrol agent with multiple plant growth-promoting properties. To our knowledge, this is the first report on the antifungal, IAA production and tomato growth enhancing compounds produced by P. lilacinum LPSC #876. Ivana A. Cavello, Juan M. Crespo, Sabrina S. García, José M. Zapiola, María F. Luna, and Sebastián F. Cavalitto Copyright © 2015 Ivana A. Cavello et al. 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.