Pub Date : 2018-02-11eCollection Date: 2018-01-01DOI: 10.1155/2018/9745198
Temesgen Oljira, Diriba Muleta, Mulissa Jida
Biological wastewater treatment is economically feasible and ecofriendly. This study was aimed at isolating bacteria from brewery wastes and evaluating their bioremediation potential as individual isolate and/or their consortium in reducing the pollutants of brewery effluents. A total of 40 bacterial isolates were recovered and of these the three best isolates were selected. The selected bacteria were identified to genus level by using morphological and biochemical characteristics. Accordingly, the isolates were identified as Aeromonas sp., Pseudomonas sp., and Bacillus sp. After 12 days of incubation, the removal efficiency of these three isolates and their combinations for biological oxygen demand and chemical oxygen demand varied from 73.55% to 94.85% and 76.78% to 93.25%, respectively. Total nitrogen and phosphorus removal was within the range of 54.43% to 77.21% and 41.80% to 78.18%, respectively. Total suspended solid, total solid, and total dissolved solids removal ranged from 66.74% to 90.3%, 54.69% to 88.5%, and 53.02% to 88.2%, respectively. The pH and electrical conductivity values ranged from 6.81 to 8.65 and 3.31 mS/cm to 3.67 mS/cm, respectively. The treated effluent increased Beta vulgaris seeds germination from 80% to 100%, with mean germination time of 3.1 to 5.2 days and seedlings length of 2.3 cm to 6.3 cm. Therefore, the development of this finding into a large scale offers an attractive technology for brewery waste treatment.
{"title":"Potential Applications of Some Indigenous Bacteria Isolated from Polluted Areas in the Treatment of Brewery Effluents.","authors":"Temesgen Oljira, Diriba Muleta, Mulissa Jida","doi":"10.1155/2018/9745198","DOIUrl":"https://doi.org/10.1155/2018/9745198","url":null,"abstract":"<p><p>Biological wastewater treatment is economically feasible and ecofriendly. This study was aimed at isolating bacteria from brewery wastes and evaluating their bioremediation potential as individual isolate and/or their consortium in reducing the pollutants of brewery effluents. A total of 40 bacterial isolates were recovered and of these the three best isolates were selected. The selected bacteria were identified to genus level by using morphological and biochemical characteristics. Accordingly, the isolates were identified as <i>Aeromonas</i> sp., <i>Pseudomonas</i> sp., and <i>Bacillus</i> sp. After 12 days of incubation, the removal efficiency of these three isolates and their combinations for biological oxygen demand and chemical oxygen demand varied from 73.55% to 94.85% and 76.78% to 93.25%, respectively. Total nitrogen and phosphorus removal was within the range of 54.43% to 77.21% and 41.80% to 78.18%, respectively. Total suspended solid, total solid, and total dissolved solids removal ranged from 66.74% to 90.3%, 54.69% to 88.5%, and 53.02% to 88.2%, respectively. The pH and electrical conductivity values ranged from 6.81 to 8.65 and 3.31 mS/cm to 3.67 mS/cm, respectively. The treated effluent increased <i>Beta vulgaris</i> seeds germination from 80% to 100%, with mean germination time of 3.1 to 5.2 days and seedlings length of 2.3 cm to 6.3 cm. Therefore, the development of this finding into a large scale offers an attractive technology for brewery waste treatment.</p>","PeriodicalId":9268,"journal":{"name":"Biotechnology Research International","volume":"2018 ","pages":"9745198"},"PeriodicalIF":0.0,"publicationDate":"2018-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2018/9745198","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35972379","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-01-01Epub Date: 2017-05-23DOI: 10.1155/2017/8791359
Jörn Josewski, Sabine Buchmeier, André Frenzel, Philip Tinnefeld, Stefan Dübel, Udo Rau
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.
来自担子菌Schizophyllum commune的β -(1,6)-分支β -(1,3)- d -葡聚糖(如schizophyllan))激发各种免疫刺激作用,并已作为佐剂进行临床试验。由于其在水溶液中的粘度和温度稳定性,一些葡聚糖也适用于食品或汽油工业。针对这些葡聚糖的抗体可作为分析葡聚糖制剂或进一步研究其生物活性的工具。因此,利用裂叶菌免疫小鼠,构建了免疫噬菌体展示文库。通过对裂叶植物的亲和选择,从该文库中分离到3个重组单克隆抗体。这些抗体的半最大有效浓度(EC50)值在16.4 ~ 21.3 ng mL-1之间变化。该克隆不仅对裂叶植物具有结合特异性,而且对具有相似大分子结构的β -(1,6)支链β -(1,3)- d -葡聚糖也具有结合特异性。二级结构的变性导致抗体结合减少,表明表位需要葡聚糖的三螺旋结构的正确构象。
{"title":"Generation of Recombinant Antibodies against the beta-(1,6)-Branched beta-(1,3)-D-Glucan Schizophyllan from Immunized Mice via Phage Display.","authors":"Jörn Josewski, Sabine Buchmeier, André Frenzel, Philip Tinnefeld, Stefan Dübel, Udo Rau","doi":"10.1155/2017/8791359","DOIUrl":"https://doi.org/10.1155/2017/8791359","url":null,"abstract":"<p><p>beta-(1,6)-Branched beta-(1,3)-D-glucans like schizophyllan from the basidiomycete <i>Schizophyllum commune</i> 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<sup>-1</sup> and 21.3 ng mL<sup>-1</sup>. 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.</p>","PeriodicalId":9268,"journal":{"name":"Biotechnology Research International","volume":"2017 ","pages":"8791359"},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1155/2017/8791359","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35092891","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2017-01-01Epub Date: 2017-12-13DOI: 10.1155/2017/1925820
Tika B Karki, Parash Mani Timilsina, Archana Yadav, Gyanu Raj Pandey, Yogesh Joshi, Sahansila Bhujel, Rojina Adhikari, Katyayanee Neupane
The study aims to isolate the yeast strains that could be used effectively as baker's yeast and compare them with the commercial baker's yeast available in the market of Nepal. A total of 10 samples including locally available sources like fruits, Murcha, and a local tree "Dar" were collected from different localities of Bhaktapur, Kavre, and Syangja districts of Nepal, respectively. Following enrichment and fermentation of the samples, 26 yeast strains were isolated using selective medium Wallerstein Laboratory Nutrient Agar. From the differential tests which included morphological and microscopic observation and physiological and biochemical characterization such as nitrate reduction and lactose utilization tests, 8 strains were selected as possible Saccharomyces strain. The selected strains were further assessed for their efficient leavening ability by tests such as ethanol tolerance, osmotolerance, invertase test, and stress exclusion test. The three most potent strains ENG, MUR3B, and SUG1 isolated from grape, Murcha, and sugarcane, respectively, were used in the fermentation and baking of dough. These strains also carried a possibility of being used as industrial baker's yeast.
{"title":"Selection and Characterization of Potential Baker's Yeast from Indigenous Resources of Nepal.","authors":"Tika B Karki, Parash Mani Timilsina, Archana Yadav, Gyanu Raj Pandey, Yogesh Joshi, Sahansila Bhujel, Rojina Adhikari, Katyayanee Neupane","doi":"10.1155/2017/1925820","DOIUrl":"10.1155/2017/1925820","url":null,"abstract":"<p><p>The study aims to isolate the yeast strains that could be used effectively as baker's yeast and compare them with the commercial baker's yeast available in the market of Nepal. A total of 10 samples including locally available sources like fruits, Murcha, and a local tree \"Dar\" were collected from different localities of Bhaktapur, Kavre, and Syangja districts of Nepal, respectively. Following enrichment and fermentation of the samples, 26 yeast strains were isolated using selective medium Wallerstein Laboratory Nutrient Agar. From the differential tests which included morphological and microscopic observation and physiological and biochemical characterization such as nitrate reduction and lactose utilization tests, 8 strains were selected as possible <i>Saccharomyces</i> strain. The selected strains were further assessed for their efficient leavening ability by tests such as ethanol tolerance, osmotolerance, invertase test, and stress exclusion test. The three most potent strains ENG, MUR3B, and SUG1 isolated from grape, Murcha, and sugarcane, respectively, were used in the fermentation and baking of dough. These strains also carried a possibility of being used as industrial baker's yeast.</p>","PeriodicalId":9268,"journal":{"name":"Biotechnology Research International","volume":"2017 ","pages":"1925820"},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5745694/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35781651","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
P. Virupakshappa, Manjunatha Bukkambudhi Krishnaswamy, G. Mishra, Mohammed Ameenuddin Mehkri
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 R 2 as 0.7678 with statistically significant P 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.
{"title":"Optimization of Crude Oil and PAHs Degradation by Stenotrophomonas rhizophila KX082814 Strain through Response Surface Methodology Using Box-Behnken Design","authors":"P. Virupakshappa, Manjunatha Bukkambudhi Krishnaswamy, G. Mishra, Mohammed Ameenuddin Mehkri","doi":"10.1155/2016/4769542","DOIUrl":"https://doi.org/10.1155/2016/4769542","url":null,"abstract":"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 R 2 as 0.7678 with statistically significant P 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.","PeriodicalId":9268,"journal":{"name":"Biotechnology Research International","volume":"10 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-12-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88856916","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
[This corrects the article DOI: 10.1155/2016/3427098.].
[这更正了文章DOI: 10.1155/2016/3427098.]。
{"title":"Corrigendum to “Proteases from Canavalia ensiformis: Active and Thermostable Enzymes with Potential of Application in Biotechnology”","authors":"Rayane Natashe Gonçalves, Suellen Duarte Gozzini Barbosa, Raquel Elisa da Silva-López","doi":"10.1155/2016/9872540","DOIUrl":"https://doi.org/10.1155/2016/9872540","url":null,"abstract":"[This corrects the article DOI: 10.1155/2016/3427098.].","PeriodicalId":9268,"journal":{"name":"Biotechnology Research International","volume":"25 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81726047","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Birgit Spitzer-Sonnleitner, A. Kempe, Maximilian Lackner
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.
{"title":"Influence of Halide Solutions on Collagen Networks: Measurements of Physical Properties by Atomic Force Microscopy","authors":"Birgit Spitzer-Sonnleitner, A. Kempe, Maximilian Lackner","doi":"10.1155/2016/4956756","DOIUrl":"https://doi.org/10.1155/2016/4956756","url":null,"abstract":"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.","PeriodicalId":9268,"journal":{"name":"Biotechnology Research International","volume":"9 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79177837","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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.
{"title":"Effect of Purified Mushroom Tyrosinase on Melanin Content and Melanogenic Protein Expression","authors":"Kamal Uddin Zaidi, Sharique A. Ali, Ayesha S Ali","doi":"10.1155/2016/9706214","DOIUrl":"https://doi.org/10.1155/2016/9706214","url":null,"abstract":"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.","PeriodicalId":9268,"journal":{"name":"Biotechnology Research International","volume":"25 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87308421","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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.
{"title":"Proteases from Canavalia ensiformis: Active and Thermostable Enzymes with Potential of Application in Biotechnology","authors":"Rayane Natashe Gonçalves, Suellen Duarte Gozzini Barbosa, Raquel Elisa da Silva-López","doi":"10.1155/2016/3427098","DOIUrl":"https://doi.org/10.1155/2016/3427098","url":null,"abstract":"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.","PeriodicalId":9268,"journal":{"name":"Biotechnology Research International","volume":"71 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-08-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81514550","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
M. Aljurf, H. Abalkhail, A. Alseraihy, S. Mohamed, M. Ayas, F. Alsharif, H. Alzahrani, A. Al-Jefri, G. Aldawsari, A. Al-Ahmari, A. Belgaumi, C. Walter, H. El-Solh, W. Rasheed, M. Albitar
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 (N = 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.
{"title":"Chimerism Analysis of Cell-Free DNA in Patients Treated with Hematopoietic Stem Cell Transplantation May Predict Early Relapse in Patients with Hematologic Malignancies","authors":"M. Aljurf, H. Abalkhail, A. Alseraihy, S. Mohamed, M. Ayas, F. Alsharif, H. Alzahrani, A. Al-Jefri, G. Aldawsari, A. Al-Ahmari, A. Belgaumi, C. Walter, H. El-Solh, W. Rasheed, M. Albitar","doi":"10.1155/2016/8589270","DOIUrl":"https://doi.org/10.1155/2016/8589270","url":null,"abstract":"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 (N = 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.","PeriodicalId":9268,"journal":{"name":"Biotechnology Research International","volume":"11 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81520275","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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.
生物表面活性剂是由细菌或酵母利用不同的底物如糖、甘油或油生产的。它们在洗涤剂、石油和制药工业中有重要的应用。甘油是生物柴油工业的产物,现有的甘油市场无法容纳过剩的产量;因此,需要开发精制甘油的新市场。本研究的目的是优化利用废甘油生产微生物鼠李糖脂。我们开发了一种生产鼠李糖脂生物表面活性剂的工艺,使用甘油作为唯一的碳源,通过广泛的筛选程序获得当地铜绿假单胞菌分离物。采用因子设计优化鼠李糖脂的产量。细胞在pH为6、含1% (v/v)甘油和2% (w/v)硝酸钠为氮源、温度为37℃、转速为180 rpm的最低盐培养基中培养2天后产量最高,培养54 h (0.04 g/L h)后产量可达2.164 g/L。通过薄层色谱、高效液相色谱和红外光谱分析,证实了该生物表面活性剂为单鼠李糖脂。甘油可以作为微生物分离物生产鼠李糖脂的来源,提供了一种廉价可靠的底物。
{"title":"Utilization of Crude Glycerol as a Substrate for the Production of Rhamnolipid by Pseudomonas aeruginosa","authors":"Walaa A. Eraqi, A. Yassin, Amal E. Ali, M. Amin","doi":"10.1155/2016/3464509","DOIUrl":"https://doi.org/10.1155/2016/3464509","url":null,"abstract":"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.","PeriodicalId":9268,"journal":{"name":"Biotechnology Research International","volume":"8 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84222756","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}