- Email: [email protected]
Effects of stirring and aeration rates on lipase production and growth of R. delemar
New Biotechnology · Volume 29S · September 2012
fact that Royal Botanic Garden, Kew is among the special gardens in the world and possesses the largest seed bank of the world shows that it has a high potential value. This study examined the differences among the socio-economic, cultural and demographic features of visitors of RBG and analyzed whether these elements are effective upon the protection of natural resources and determination of economic value. Examining the results of 460 surveys that were performed in Royal Botanic Garden Kew; it was determined that 39% are male and 61% are female visitors. According to the survey results, it was determined that people from different age groups visit the area. Examining the visitors in terms of age groups; it was determined that the age range of 26–65 predominates. While 51% of the study participants have a degree of technical university, 32% have a degree of post-graduate. 32% of the visitors work in private sector, 20% have an income of 20,000–30,000 and 22% have an income of 50,000 and above. It was determined that 81% of the visitors are not associated with any NGOs. Keywords: Role of botanic gardens; Visitor’s preferences; Protection value; Royal Botanic Garden-Kew http://dx.doi.org/10.1016/j.nbt.2012.08.546 Poster 5.0.107 Bioaugmentation of biobed biomixture with fungal pellets of Anthracophyllum discolor to improve pesticides degradation M.C. Diez1,2,∗ , S. Elgueta2 , S. Fernandez2 , F. Gallardo2 1
Chemical Engineering Department, Universidad de La Frontera, PO Box 54-D, Temuco, Chile 2 Scientific and Technological Bioresource Nucleus, Universidad de La Frontera, PO Box 54-D, Temuco, Chile Biobed is a biopurification system used to minimize environmental contamination during manipulation of pesticides, especially when filling the spraying equipment. Biobeds technology is based on the adsorption and degradation of pesticides by a biomixture composed of top soil, peat, and straw. Their efficiency could be improved by bioaugmentation with specific microorganisms. Therefore, we studied the bioaugmentation of biomixture with fungal pellets of Anthracophyllum discolor, a white-rot fungus with high ligninolytic activity, on pesticides degradation and on biomixture biological activities. The biomixture was prepared mixing top soil (Andisol), straw and peat (1:2:1, v/v/v) and was inoculated with the fungal pellets (10%, w/w). The biomixture was contaminated with 100 mg kg−1 of a mix of pesticides (atrazine, chlorpyrifos, iprodione and isoproturon) and with each single pesticide. Biomixtures were incubated during 90 days at 20◦ C and samples were extracted periodically to evaluate residual pesticides and biological activities (FDA, respiratory activity. and phenoloxidases). The biomixture inoculated and contaminated with a mix of 4 pesticides showed higher and faster degradation than noninoculated biomixture, except for isoproturon. Atrazine was 100% degraded even after 30 days, 18% faster than the biomixture noninoculated. After 90th days of incubation, the FDA activity was highest for single pesticide, especially atrazine (17 g FDA g−1 h−1 ); however, the respiratory activity was highest for the mix of 4 pes-
ticides (4118 mg CO2 g−1 ) and the highest phenoloxidases activity was for single chlorpyrifos (3.3 U kg−1 ). Acknowledgement: Financed partially by Fondecyt 1120963 and FONDEF D09R1006 projects. http://dx.doi.org/10.1016/j.nbt.2012.08.547 Poster 5.0.108 Comparison of heavy metal uptake capacities of growing, resting and dead cells of R. delemar via acid phosphatase activity Mehtap Ers¸an∗ , Ünsal Ac¸ıkel Department of Chemical Engineering, Cumhuriyet University, Sivas, Turkey The bioaccumulation of heavy metals via acid phosphatase activity and growth of R. delemar were investigated. The phosphatase activity and growth of R. delemar were generally inhibited in the presence of increasing concentrations in the range 10–100 mg/L of heavy metals. Heavy metal removal capacities of growing, resting, dead cells of R. delemar were compared. Maximum Ni(II) removal efficiencies by growing, resting, dead cells of R. delemar in the presence of 10 mg/L Ni(II) were found to be 52.8%, 36.1%, 36.9%, respectively. In case of Cu(II), maximum Cu(II) removal efficiencies by growing, resting, dead cells of R. delemar in the presence of 10 mg/L Cu(II) were determined as 79.5%, 49.7%, 48.8%, respectively. The inhibition effect of Zn(II) on the phosphatase activity of R. delemar began to be effective at an initial Zn(II) concentration of 50 mg/L. Maximum Zn(II) removal efficiencies by growing, resting, dead cells of R. delemar were obtained as 80.8% at an initial concentration of 50 mg/L, 48.1% and 46.4% at initial concentration of 10 mg/L, respectively. The heavy metal sorption capacities of growing, resting, dead cells of R. delemar were evaluated in terms of sorption isotherms and the best correlation was obtained using the Freundlich model. http://dx.doi.org/10.1016/j.nbt.2012.08.548 Poster 5.0.109 Effects of stirring and aeration rates on lipase production and growth of R. delemar g Ac¸ıkel2 Mehtap Ers¸an1,∗ , Ünsal Ac¸ıkel1 , Yes¸im Sa˘ 1
Department of Chemical Engineering, Cumhuriyet University, Sivas, Turkey 2 Department of Chemical Engineering, Hacettepe University, Beytepe, Ankara, Turkey Lipases (triacylglycerol hydrolases) are hydrolytic enzymes that can catalyze the hydrolysis of the ester bond of long-chain acylglycerols at the oil–water interface. The presence of air was essential for lipase production by R. delemar. In this study, the effects of stirring rate, aeration rate, and the amount of perfluorocarbon(PFR), an oxygen carrier, in two different nutrient media containing glucose and sucrose as main carbon sources were investigated in a batch stirred reactor. A maximum lipase activity of 103.12 mol/L min was obtained at 200 rpm stirring rate. The
www.elsevier.com/locate/nbt S195
New Biotechnology · Volume 29S · September 2012
effect of aeration rate on the lipase activity was investigated in the range of 1–4 vvm. The maximum lipase activities in the media containing glucose and sucrose as main C sources were found to be 61.07 mol/L min and 102.83 mol/L min, respectively at the stationary phase of growth (96th hour of fermentation) at 3 vvm and 2 vvm aeration rates, respectively. The inclusion of 10% PFR in the fermentation medium resulted in higher lipase production. The maximum lipase activities in the media containing glucose and sucrose were determined as102.67 mol/L min and 171.20 mol/L min, respectively at 2 vvm aeration rate. http://dx.doi.org/10.1016/j.nbt.2012.08.549 Poster 5.0.110 Modification of explant’s metabolic activity to increase regeneration capacity of flax (Linum usitatissimum L.) hypocotyl segments Cansu Telci Kahramanogullari1 , Behrouz Alizadeh2 , Ramazan Beyaz3 , Mustafa Yildiz1,∗ 1
University of Ankara, Faculty of Agriculture, Department of Field Crops 06110 Dıs¸kapı, Ankara, Turkey 2 University of Ankara, Graduate School of Natural and Applied Sciences, Department of Field Crops, Gölbas¸ı, Ankara, Turkey 3 University of Ankara, Biotechnology Institute, Gölbas¸ı, Ankara, Turkey This study was conducted to investigate the effects of removing one of the cotyledon leaves of in vitro-grown flax (Linum usitatissimum L.) seedling on regeneration capacity of hypocotyls. Seeds from two flax cvs. ‘Madaras’ and ‘1886 Sel.’ were surface sterilized with 40% commercial bleach containing 5% sodium hypochlorite at 10◦ C for 12 min with continuous stirring and then were washed three times with sterile distilled water at the same temperature. Sterilized seeds were germinated on a basal medium containing the mineral salts and vitamins of Murashige and Skoog (MS), 3% (w/v) sucrose and 0.7% (w/v) agar in Magenta vessels. One of the cotyledon leaves of some seedling was removed 5 days after culture initiation while other seedlings kept as they were. 14 days after seed sown for germination, seedling and root lengths, seedling fresh and dry weights, seedling water and dry matter contents, chlorophyll a, chlorophyll b and total chlorophyll contents in cotyledon leaves of seedlings were recorded. Hypocotyl segments were cultured for regeneration for 4 weeks. Sixteen explants were cultured at 1.0 cm × 1.0 cm distance with a 4 × 4 matrix in a Petri dish. At the end of the culture, shoot regeneration percentage, shoot number per explant, regenerated shoot length and total shoot number per Petri dish were recorded in hypocotyls excised from complete- and one cotyledon-absent-seedlings. According to the results, it could be concluded that removing one cotyledon leaf forced seedlings to increase metabolic activity to sustain their viability as seedlings with two cotyledons. Keywords: Flax; Metabolic activity; Regeneration capacity http://dx.doi.org/10.1016/j.nbt.2012.08.550
S196
www.elsevier.com/locate/nbt
Poster 5.0.111 Recent advances in microbial degradation of sulfur, nitrogen and oxygen heterocycles Ping Xu1,2 1
State Key Laboratory of Microbial Metabolism & School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People’s Republic of China 2 State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, Shandong Province, People’s Republic of China Sulfur (S), nitrogen (N) and oxygen (O) heterocycles are among the most potent environmental pollutants. Microbial degradation of these pollutants is attracting more and more attention because such bioprocesses are environmentally friendly. The biotechnological potential of these processes is being investigated, for example, to achieve better sulfur removal by immobilized biocatalysts with magnetite nanoparticles or by solvent-tolerant bacteria, and to obtain valuable intermediates from these heterocycles. Our recent advances have demonstrated the mechanism of the important nitrogen heterocycle-nicotine degradation by Pseudomonas. However, these technologies are not yet available for large-scale applications so future research must investigate proper modifications for industrial applications of these processes. This presentation focuses on recent progress in understanding how microbes degrade S, N and O heterocycles. Supporting publications: Tang HZ et al., 2012, Sci. Rep. 2:377; Li A et al., Biotechnol. Bioeng. 2012, 109:609–613; Tang HZ et al., J. Biol. Chem. 2011, 286:39179; Tang HZ et al., J. Bacteriol. 2011, 193:6789–6790; Yu H et al., J. Bacteriol. 2011, 193:5541–5542; Gai ZH et al., PLoS ONE 2010, 5:e10018; Li QG et al., Environ. Sci. Technol. 2009, 43:8635–8642; Tang HZ et al., Appl. Environ. Microbiol. 2009, 75:772–778; Tang HZ et al., Appl. Environ. Microbiol. 2008, 74:1567–1574; Xu P. et al., Trends Microbiol. 2006, 14:398; Wang SN et al., Environ. Sci. Technol. 2005, 39:6877–6880. http://dx.doi.org/10.1016/j.nbt.2012.08.551 Poster 5.0.112 Verification of GMO analytical methods: DNA isolation and quantitative detection methods for Roundup Ready soy flour Remziye Yılmaz∗ , Ceren Bayrac¸, Pelin Mutlu, Erdem Boy, Meral Yücel Middle East Technical University, Molecular Biology and Biotechnology R&D Center, Genome Analysis Laboratory, 06800 Ankara, Turkey Quality assurance is a prerequisite for accurate and reliable results in food and feed testing, ISO/IEC 17025 being recognized worldwide as the base standard. Accreditation is a suitable system for harmonising procedures in each testing laboratory. Precisely defined procedures for the verification of methods are the key points for accreditation. Testing for genetically modified organisms (GMOs) is a challenging exercise, especially with more GMOs entering the world market. We describe here the organization and performance of verifying DNA isolation and quantitative detection methods for GMO testing of Roundup Ready soy flour in the con-
fact that Royal Botanic Garden, Kew is among the special gardens in the world and possesses the largest seed bank of the world shows that it has a high potential value. This study examined the differences among the socio-economic, cultural and demographic features of visitors of RBG and analyzed whether these elements are effective upon the protection of natural resources and determination of economic value. Examining the results of 460 surveys that were performed in Royal Botanic Garden Kew; it was determined that 39% are male and 61% are female visitors. According to the survey results, it was determined that people from different age groups visit the area. Examining the visitors in terms of age groups; it was determined that the age range of 26–65 predominates. While 51% of the study participants have a degree of technical university, 32% have a degree of post-graduate. 32% of the visitors work in private sector, 20% have an income of 20,000–30,000 and 22% have an income of 50,000 and above. It was determined that 81% of the visitors are not associated with any NGOs. Keywords: Role of botanic gardens; Visitor’s preferences; Protection value; Royal Botanic Garden-Kew http://dx.doi.org/10.1016/j.nbt.2012.08.546 Poster 5.0.107 Bioaugmentation of biobed biomixture with fungal pellets of Anthracophyllum discolor to improve pesticides degradation M.C. Diez1,2,∗ , S. Elgueta2 , S. Fernandez2 , F. Gallardo2 1
Chemical Engineering Department, Universidad de La Frontera, PO Box 54-D, Temuco, Chile 2 Scientific and Technological Bioresource Nucleus, Universidad de La Frontera, PO Box 54-D, Temuco, Chile Biobed is a biopurification system used to minimize environmental contamination during manipulation of pesticides, especially when filling the spraying equipment. Biobeds technology is based on the adsorption and degradation of pesticides by a biomixture composed of top soil, peat, and straw. Their efficiency could be improved by bioaugmentation with specific microorganisms. Therefore, we studied the bioaugmentation of biomixture with fungal pellets of Anthracophyllum discolor, a white-rot fungus with high ligninolytic activity, on pesticides degradation and on biomixture biological activities. The biomixture was prepared mixing top soil (Andisol), straw and peat (1:2:1, v/v/v) and was inoculated with the fungal pellets (10%, w/w). The biomixture was contaminated with 100 mg kg−1 of a mix of pesticides (atrazine, chlorpyrifos, iprodione and isoproturon) and with each single pesticide. Biomixtures were incubated during 90 days at 20◦ C and samples were extracted periodically to evaluate residual pesticides and biological activities (FDA, respiratory activity. and phenoloxidases). The biomixture inoculated and contaminated with a mix of 4 pesticides showed higher and faster degradation than noninoculated biomixture, except for isoproturon. Atrazine was 100% degraded even after 30 days, 18% faster than the biomixture noninoculated. After 90th days of incubation, the FDA activity was highest for single pesticide, especially atrazine (17 g FDA g−1 h−1 ); however, the respiratory activity was highest for the mix of 4 pes-
ticides (4118 mg CO2 g−1 ) and the highest phenoloxidases activity was for single chlorpyrifos (3.3 U kg−1 ). Acknowledgement: Financed partially by Fondecyt 1120963 and FONDEF D09R1006 projects. http://dx.doi.org/10.1016/j.nbt.2012.08.547 Poster 5.0.108 Comparison of heavy metal uptake capacities of growing, resting and dead cells of R. delemar via acid phosphatase activity Mehtap Ers¸an∗ , Ünsal Ac¸ıkel Department of Chemical Engineering, Cumhuriyet University, Sivas, Turkey The bioaccumulation of heavy metals via acid phosphatase activity and growth of R. delemar were investigated. The phosphatase activity and growth of R. delemar were generally inhibited in the presence of increasing concentrations in the range 10–100 mg/L of heavy metals. Heavy metal removal capacities of growing, resting, dead cells of R. delemar were compared. Maximum Ni(II) removal efficiencies by growing, resting, dead cells of R. delemar in the presence of 10 mg/L Ni(II) were found to be 52.8%, 36.1%, 36.9%, respectively. In case of Cu(II), maximum Cu(II) removal efficiencies by growing, resting, dead cells of R. delemar in the presence of 10 mg/L Cu(II) were determined as 79.5%, 49.7%, 48.8%, respectively. The inhibition effect of Zn(II) on the phosphatase activity of R. delemar began to be effective at an initial Zn(II) concentration of 50 mg/L. Maximum Zn(II) removal efficiencies by growing, resting, dead cells of R. delemar were obtained as 80.8% at an initial concentration of 50 mg/L, 48.1% and 46.4% at initial concentration of 10 mg/L, respectively. The heavy metal sorption capacities of growing, resting, dead cells of R. delemar were evaluated in terms of sorption isotherms and the best correlation was obtained using the Freundlich model. http://dx.doi.org/10.1016/j.nbt.2012.08.548 Poster 5.0.109 Effects of stirring and aeration rates on lipase production and growth of R. delemar g Ac¸ıkel2 Mehtap Ers¸an1,∗ , Ünsal Ac¸ıkel1 , Yes¸im Sa˘ 1
Department of Chemical Engineering, Cumhuriyet University, Sivas, Turkey 2 Department of Chemical Engineering, Hacettepe University, Beytepe, Ankara, Turkey Lipases (triacylglycerol hydrolases) are hydrolytic enzymes that can catalyze the hydrolysis of the ester bond of long-chain acylglycerols at the oil–water interface. The presence of air was essential for lipase production by R. delemar. In this study, the effects of stirring rate, aeration rate, and the amount of perfluorocarbon(PFR), an oxygen carrier, in two different nutrient media containing glucose and sucrose as main carbon sources were investigated in a batch stirred reactor. A maximum lipase activity of 103.12 mol/L min was obtained at 200 rpm stirring rate. The
www.elsevier.com/locate/nbt S195
New Biotechnology · Volume 29S · September 2012
effect of aeration rate on the lipase activity was investigated in the range of 1–4 vvm. The maximum lipase activities in the media containing glucose and sucrose as main C sources were found to be 61.07 mol/L min and 102.83 mol/L min, respectively at the stationary phase of growth (96th hour of fermentation) at 3 vvm and 2 vvm aeration rates, respectively. The inclusion of 10% PFR in the fermentation medium resulted in higher lipase production. The maximum lipase activities in the media containing glucose and sucrose were determined as102.67 mol/L min and 171.20 mol/L min, respectively at 2 vvm aeration rate. http://dx.doi.org/10.1016/j.nbt.2012.08.549 Poster 5.0.110 Modification of explant’s metabolic activity to increase regeneration capacity of flax (Linum usitatissimum L.) hypocotyl segments Cansu Telci Kahramanogullari1 , Behrouz Alizadeh2 , Ramazan Beyaz3 , Mustafa Yildiz1,∗ 1
University of Ankara, Faculty of Agriculture, Department of Field Crops 06110 Dıs¸kapı, Ankara, Turkey 2 University of Ankara, Graduate School of Natural and Applied Sciences, Department of Field Crops, Gölbas¸ı, Ankara, Turkey 3 University of Ankara, Biotechnology Institute, Gölbas¸ı, Ankara, Turkey This study was conducted to investigate the effects of removing one of the cotyledon leaves of in vitro-grown flax (Linum usitatissimum L.) seedling on regeneration capacity of hypocotyls. Seeds from two flax cvs. ‘Madaras’ and ‘1886 Sel.’ were surface sterilized with 40% commercial bleach containing 5% sodium hypochlorite at 10◦ C for 12 min with continuous stirring and then were washed three times with sterile distilled water at the same temperature. Sterilized seeds were germinated on a basal medium containing the mineral salts and vitamins of Murashige and Skoog (MS), 3% (w/v) sucrose and 0.7% (w/v) agar in Magenta vessels. One of the cotyledon leaves of some seedling was removed 5 days after culture initiation while other seedlings kept as they were. 14 days after seed sown for germination, seedling and root lengths, seedling fresh and dry weights, seedling water and dry matter contents, chlorophyll a, chlorophyll b and total chlorophyll contents in cotyledon leaves of seedlings were recorded. Hypocotyl segments were cultured for regeneration for 4 weeks. Sixteen explants were cultured at 1.0 cm × 1.0 cm distance with a 4 × 4 matrix in a Petri dish. At the end of the culture, shoot regeneration percentage, shoot number per explant, regenerated shoot length and total shoot number per Petri dish were recorded in hypocotyls excised from complete- and one cotyledon-absent-seedlings. According to the results, it could be concluded that removing one cotyledon leaf forced seedlings to increase metabolic activity to sustain their viability as seedlings with two cotyledons. Keywords: Flax; Metabolic activity; Regeneration capacity http://dx.doi.org/10.1016/j.nbt.2012.08.550
S196
www.elsevier.com/locate/nbt
Poster 5.0.111 Recent advances in microbial degradation of sulfur, nitrogen and oxygen heterocycles Ping Xu1,2 1
State Key Laboratory of Microbial Metabolism & School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People’s Republic of China 2 State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, Shandong Province, People’s Republic of China Sulfur (S), nitrogen (N) and oxygen (O) heterocycles are among the most potent environmental pollutants. Microbial degradation of these pollutants is attracting more and more attention because such bioprocesses are environmentally friendly. The biotechnological potential of these processes is being investigated, for example, to achieve better sulfur removal by immobilized biocatalysts with magnetite nanoparticles or by solvent-tolerant bacteria, and to obtain valuable intermediates from these heterocycles. Our recent advances have demonstrated the mechanism of the important nitrogen heterocycle-nicotine degradation by Pseudomonas. However, these technologies are not yet available for large-scale applications so future research must investigate proper modifications for industrial applications of these processes. This presentation focuses on recent progress in understanding how microbes degrade S, N and O heterocycles. Supporting publications: Tang HZ et al., 2012, Sci. Rep. 2:377; Li A et al., Biotechnol. Bioeng. 2012, 109:609–613; Tang HZ et al., J. Biol. Chem. 2011, 286:39179; Tang HZ et al., J. Bacteriol. 2011, 193:6789–6790; Yu H et al., J. Bacteriol. 2011, 193:5541–5542; Gai ZH et al., PLoS ONE 2010, 5:e10018; Li QG et al., Environ. Sci. Technol. 2009, 43:8635–8642; Tang HZ et al., Appl. Environ. Microbiol. 2009, 75:772–778; Tang HZ et al., Appl. Environ. Microbiol. 2008, 74:1567–1574; Xu P. et al., Trends Microbiol. 2006, 14:398; Wang SN et al., Environ. Sci. Technol. 2005, 39:6877–6880. http://dx.doi.org/10.1016/j.nbt.2012.08.551 Poster 5.0.112 Verification of GMO analytical methods: DNA isolation and quantitative detection methods for Roundup Ready soy flour Remziye Yılmaz∗ , Ceren Bayrac¸, Pelin Mutlu, Erdem Boy, Meral Yücel Middle East Technical University, Molecular Biology and Biotechnology R&D Center, Genome Analysis Laboratory, 06800 Ankara, Turkey Quality assurance is a prerequisite for accurate and reliable results in food and feed testing, ISO/IEC 17025 being recognized worldwide as the base standard. Accreditation is a suitable system for harmonising procedures in each testing laboratory. Precisely defined procedures for the verification of methods are the key points for accreditation. Testing for genetically modified organisms (GMOs) is a challenging exercise, especially with more GMOs entering the world market. We describe here the organization and performance of verifying DNA isolation and quantitative detection methods for GMO testing of Roundup Ready soy flour in the con-