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Abstracts / Journal of Biotechnology 208 (2015) S5–S120
Studies on purification of capsaicinoids from various Capsicum species cultivated in Romania Roxana M˘ad˘alina Stoica ∗ , Narcisa B˘abeanu, Ovidiu Popa Faculty of Biotechnologies, University of Agronomic Sciences and Veterinary Medicine of Bucharest, Romania E-mail address: roxym [email protected] (R.M. Stoica). Plants produce a diversity of low molecular secondary metabolites. The most important components in the group of secondary metabolites derived from Capsicum species are capsaicinoids. The purpose of our study was to evaluate the total content of capsaicinoids in three varieties of pepper (Capsicum annuum L.), such as Fiesta, Fresno and Hot Cherry cultivars, which were grown in Romania agro-climatic conditions, and also, to check the influence of particle diameter on the extraction yield. Extraction was done using several organic solvents (ethanol, methanol, acetone and acetonitrile) and milled pepper were separated in three granulometric classes: 0.8–1 m, 0.5–0.8 m and <0.5 m. The solvents used for the extraction showed different capsaicinoids extraction yield. So, ethanol extract showed the higher content of capsaicinoids comparing with other solvents (0.82%), whereas methanol extract showed the smallest content of capsaicinoids (0.61%). The influence of particle sizes on extraction yield was not statistically significant. Acknowledgements: This paper was published under the frame of European Social Fund, Human Resources Development Operational Programme 2007–2013, project no. POSDRU/159/1.5/S/132765. http://dx.doi.org/10.1016/j.jbiotec.2015.06.322 Complex characterization of native Anethum graveolens volatile oil Alina Ortan 1 , Alexandru Ioan Calin 1,∗ , Cristina Dinu Pirvu 2 1 University of Agronomic Science and Veterinary Medicine, Bucharest, Romania 2 University of Medicine and Pharmacy “Carol Davila”, Bucharest, Romania
E-mail address: [email protected] (A.I. Calin). Changing in consumer demands, increasing of public interest, awareness of health requirements and, not at least, the possibility of using volatile oils in biotechnology have opened a new page in the modern fitomedicine. Therefore, in many countries and communities quality requirements of volatile oils are regulated in pharmacopoeia and monographs. The aim of this study was the characterization of native Anethum graveolens volatile oil using modern analytical techniques. UV–vis and GC/MS analysis confirmed that, from 62 identified compounds, phenolic compounds, terpenoids and flavonoid derivatives are dominant. The phytochemical quantification procedures were used for the determination of total terpenoids, total flavonoids, and total phenolics content. The antioxidant activity was determined following two protocols (the DPPH assay and the chemiluminescence assay), showing that analysed volatile oil presents a significant total antioxidant capacity for all tested concentrations. The antifungal effect against C. albicans strain was evaluated using the disc diffusion method and showed that all the samples present a significant effect on the growth of the targeted fungal line.
S103
The performed tests on native Anethum graveolens volatile oil recommend it as a potential candidate for antimicrobial phytopreparates with low toxicity and convenient therapeutic effect. This work was supported by the Romanian UEFISCDI, project 176/01/07/2014 (PN-II-PT-PCCA-2013-4-0953). http://dx.doi.org/10.1016/j.jbiotec.2015.06.323 Optimization of submerged culture conditions for Flammulina velutipes on SBD culture medium Georgeta Fidler 1,∗ , Steliana Rodino 1 , Alina Butu 2 , Marian Butu 2 , Gabriela Popa 1 , Calina Petruta Cornea 1 1
University of Agronomic Sciences and Veterinary Medicine, Bucharest, Romania 2 National Institute of Research and Development for Biological Sciences, Bucharest, Romania E-mail address: gina [email protected] (G. Fidler). Flammulina velutipes is an edible and medicinal mushroom, able to produce large amounts of various bioactive compounds. Its activities are related to inhibition of pathogenic bacteria, antioxidant properties, antitumor action and immunomodulatory properties. Local isolate of this mushroom was selected in previous experiments and cultivated in low cost culture medium based on the digestate resulted from bioethanol production from Sorghum bicolor biomass (SBD). The aim of this study was the optimization of culture conditions of F. velutipes, in submerse system, using SBD medium in order to obtain increased quantities of biomass and specific bioactive compounds. Several parameters of cultivation like pH, stirring rate, addition of micro and macro elements, were evaluated. It was observed that F. velutipes is able grow within a large scale of pH, but best results were obtained at pH 5.5–6.0. Significant increase of mycelial biomass was obtained on medium supplemented with glucose, peptone, KH2 PO4 , K2 HPO4 , MgSO4 . The biomass obtained was used for extraction and tested for antioxidant and antimicrobial activities. The results were obtained under the frame of European Social Fund, Human Resources Development Operational Programme 2007–2013, project no. POSDRU/159/1.5/S/132765. http://dx.doi.org/10.1016/j.jbiotec.2015.06.324 Scaling of the HPLC method for production of the new immunostimulating phytogenesis preparations “Asgipan”, “Glabiloks” and “Sapanox” Pavel G. Alexyuk ∗ , Andrew P. Bogoyavlenskiy, Madina S. Alexyuk, Vladimir E. Berezin Department of Virology, Institute of Microbiology and Virology, Almaty, Kazakhstan E-mail address: [email protected] (P.G. Alexyuk). More and more researches to be carried out in the direction of search of the new medicines based on plant biologically active connections for treatment with infectious, oncological and somatic diseases. In turn, for the mass production of new herbal medicines needed switched from laboratory methods of obtaining active compounds to the technology of industrial production.
S104
Abstracts / Journal of Biotechnology 208 (2015) S5–S120
Earlier it was shown that vegetable preparations “Asgipan”, “Glabiloks” and “Sapanoks” are capable to increase of various vaccine preparations efficiency at various ways of administration. The purpose of this research is to develop scaling technology of laboratory methods of obtaining plant immunostimulatory preparations “Asgipan”, “Glabiloks” and “Sapanox”. Studied herbal preparations obtained by HPLC method from Aesculus hippocastanum, Glycyrrhiza glabra and Saponaria officinalis plant extracts. Scaling of HPLC process carried out in two ways: by increasing the amount of sorbent in preparative column and by varying the time of chromatography, gradient elution, the volume of eluent applied to the column per unit time. During the process of scaling carried out a permanent spectrometric control for the effectiveness of purification and quality of plant preparations “Asgipan”, “Glabiloks” and “Sapanox” was carried out. Studies have revealed that the change of eluent volume from 0.8 ml/min to 5 ml/min and the column type allow to increase the yield of the final product more than 12 times, due to the effective separation of the original plant increased in 6 times used for purification of “Asgipan”, “Glabiloks” and “Sapanox”. http://dx.doi.org/10.1016/j.jbiotec.2015.06.325 Evaluation of the biologic activity of some quaternary ammonium salts against non-pathogenic and pathogenic microorganisms Georgiana Horincar 1 , Ioana Otilia Ghinea 2 , Bianca Furdui 2 , Andreea Veronica Dediu 1 , Gabriela Bahrim 1 , Rodica Mihaela Dinica 2,∗ 1 Department of Food Science, Food Engineering and Applied Biotechnology, “Dunarea de Jos” University, Galati, Romania 2 Department of Chemistry, Physics and Environment, “Dunarea de Jos” University, Galati, Romania
E-mail address: [email protected] (R.M. Dinica). We present herein the antimicrobial activity on non-pathogenic and pathogenic microorganisms of some heterocyclic compounds, synthesized by us using non-conventional “green” methods, which are one of the most used classes of disinfectants, with wide application in hospital environments, water treatment, textiles, paint and food industry. The study on toxicity of heterocyclic compounds was performed by determining the biological activity on non-pathogenic and pathogenic microorganisms through the diffusion method and by studying the cell survival assay using 3-(4,5-dimethylthiazol2-yl)-2,5-diphenyltetrazolium bromide (MTT). MTT measures the mitochondrial function, being widely used to measure the cytotoxic potential of a chemical compound. The antimicrobial activity was strongest against Staphylococcus sp., Streptococcus sp., and E. coli, and the lowest was against Candida albicans. Some compounds, whose Staphylococcus and Streptococcus MIC are very small (≤250 mg/mL), may be of therapeutic interest. Acknowledgements: This work was supported by a grant of the Romanian National Authority for Scientific Research, CNCSUEFISCDI project number PN-II-ID-PCE-2011-3-0226. http://dx.doi.org/10.1016/j.jbiotec.2015.06.326
The susceptibility of chicken embryo to endotoxin solution eliminated by carbonized material Kuanysh T. Tastambek 1,∗ , Nuraly S. Akimbekov 1 , Ilya Digel 2 , Azhar A. Zhubanova 1 , Irina S. Savitskaya 1 , Gulzhamal Zh Abdieva 1 , Perizat S. Ualyeva 1 1 Al-Farabi Kazkah National University, Almaty, Kazakhstan 2 Aachen University of Applied Sciences, Julich, Germany
E-mail address: [email protected] (K.T. Tastambek). Bacterial endotoxin (LPS) removal from aqueous solutions is a challenging bioengineering task because of its ubiquity and high physical and chemical stability. Nowadays, adsorption methods of LPS removal are regarded as most advantageous with many different adsorbent materials being developed. In this work, low-cost nanostructured carbonized material (rice husks-CRH) designed by us in accordance for LPS-adsorption. The purpose of the present investigation was to determine the susceptibility of chicken embryos to an injection of endotoxin and CRH-treated endotoxin in vivo during the period of development. Endotoxin extracted from cell walls of E. coli by LPS extraction kid was injected intravenously into fertile eggs, of varying ages. It has been observed that the susceptibility (the development of provisional organs, pyrogenicity, blood analysis and lethal dose (LD50)) of chicken embryos to endotoxin decreased daily as the embryo matured from day 10 to hatching, while CRH-treated LPS had showed weak effect. These data show that the toxin effects of LPS solution was eliminated by CRH, according to the increased resistance of embryos. We consider the obtained results on the basis of novel advanced and economically efficient material CRH are very promising as the increasing number of biotechnological and biomedical applications supposed to be endotoxin-free, as well as still persisting problems with purification of complex mixtures. http://dx.doi.org/10.1016/j.jbiotec.2015.06.327
Studies on purification of capsaicinoids from various Capsicum species cultivated in Romania Roxana M˘ad˘alina Stoica ∗ , Narcisa B˘abeanu, Ovidiu Popa Faculty of Biotechnologies, University of Agronomic Sciences and Veterinary Medicine of Bucharest, Romania E-mail address: roxym [email protected] (R.M. Stoica). Plants produce a diversity of low molecular secondary metabolites. The most important components in the group of secondary metabolites derived from Capsicum species are capsaicinoids. The purpose of our study was to evaluate the total content of capsaicinoids in three varieties of pepper (Capsicum annuum L.), such as Fiesta, Fresno and Hot Cherry cultivars, which were grown in Romania agro-climatic conditions, and also, to check the influence of particle diameter on the extraction yield. Extraction was done using several organic solvents (ethanol, methanol, acetone and acetonitrile) and milled pepper were separated in three granulometric classes: 0.8–1 m, 0.5–0.8 m and <0.5 m. The solvents used for the extraction showed different capsaicinoids extraction yield. So, ethanol extract showed the higher content of capsaicinoids comparing with other solvents (0.82%), whereas methanol extract showed the smallest content of capsaicinoids (0.61%). The influence of particle sizes on extraction yield was not statistically significant. Acknowledgements: This paper was published under the frame of European Social Fund, Human Resources Development Operational Programme 2007–2013, project no. POSDRU/159/1.5/S/132765. http://dx.doi.org/10.1016/j.jbiotec.2015.06.322 Complex characterization of native Anethum graveolens volatile oil Alina Ortan 1 , Alexandru Ioan Calin 1,∗ , Cristina Dinu Pirvu 2 1 University of Agronomic Science and Veterinary Medicine, Bucharest, Romania 2 University of Medicine and Pharmacy “Carol Davila”, Bucharest, Romania
E-mail address: [email protected] (A.I. Calin). Changing in consumer demands, increasing of public interest, awareness of health requirements and, not at least, the possibility of using volatile oils in biotechnology have opened a new page in the modern fitomedicine. Therefore, in many countries and communities quality requirements of volatile oils are regulated in pharmacopoeia and monographs. The aim of this study was the characterization of native Anethum graveolens volatile oil using modern analytical techniques. UV–vis and GC/MS analysis confirmed that, from 62 identified compounds, phenolic compounds, terpenoids and flavonoid derivatives are dominant. The phytochemical quantification procedures were used for the determination of total terpenoids, total flavonoids, and total phenolics content. The antioxidant activity was determined following two protocols (the DPPH assay and the chemiluminescence assay), showing that analysed volatile oil presents a significant total antioxidant capacity for all tested concentrations. The antifungal effect against C. albicans strain was evaluated using the disc diffusion method and showed that all the samples present a significant effect on the growth of the targeted fungal line.
S103
The performed tests on native Anethum graveolens volatile oil recommend it as a potential candidate for antimicrobial phytopreparates with low toxicity and convenient therapeutic effect. This work was supported by the Romanian UEFISCDI, project 176/01/07/2014 (PN-II-PT-PCCA-2013-4-0953). http://dx.doi.org/10.1016/j.jbiotec.2015.06.323 Optimization of submerged culture conditions for Flammulina velutipes on SBD culture medium Georgeta Fidler 1,∗ , Steliana Rodino 1 , Alina Butu 2 , Marian Butu 2 , Gabriela Popa 1 , Calina Petruta Cornea 1 1
University of Agronomic Sciences and Veterinary Medicine, Bucharest, Romania 2 National Institute of Research and Development for Biological Sciences, Bucharest, Romania E-mail address: gina [email protected] (G. Fidler). Flammulina velutipes is an edible and medicinal mushroom, able to produce large amounts of various bioactive compounds. Its activities are related to inhibition of pathogenic bacteria, antioxidant properties, antitumor action and immunomodulatory properties. Local isolate of this mushroom was selected in previous experiments and cultivated in low cost culture medium based on the digestate resulted from bioethanol production from Sorghum bicolor biomass (SBD). The aim of this study was the optimization of culture conditions of F. velutipes, in submerse system, using SBD medium in order to obtain increased quantities of biomass and specific bioactive compounds. Several parameters of cultivation like pH, stirring rate, addition of micro and macro elements, were evaluated. It was observed that F. velutipes is able grow within a large scale of pH, but best results were obtained at pH 5.5–6.0. Significant increase of mycelial biomass was obtained on medium supplemented with glucose, peptone, KH2 PO4 , K2 HPO4 , MgSO4 . The biomass obtained was used for extraction and tested for antioxidant and antimicrobial activities. The results were obtained under the frame of European Social Fund, Human Resources Development Operational Programme 2007–2013, project no. POSDRU/159/1.5/S/132765. http://dx.doi.org/10.1016/j.jbiotec.2015.06.324 Scaling of the HPLC method for production of the new immunostimulating phytogenesis preparations “Asgipan”, “Glabiloks” and “Sapanox” Pavel G. Alexyuk ∗ , Andrew P. Bogoyavlenskiy, Madina S. Alexyuk, Vladimir E. Berezin Department of Virology, Institute of Microbiology and Virology, Almaty, Kazakhstan E-mail address: [email protected] (P.G. Alexyuk). More and more researches to be carried out in the direction of search of the new medicines based on plant biologically active connections for treatment with infectious, oncological and somatic diseases. In turn, for the mass production of new herbal medicines needed switched from laboratory methods of obtaining active compounds to the technology of industrial production.
S104
Abstracts / Journal of Biotechnology 208 (2015) S5–S120
Earlier it was shown that vegetable preparations “Asgipan”, “Glabiloks” and “Sapanoks” are capable to increase of various vaccine preparations efficiency at various ways of administration. The purpose of this research is to develop scaling technology of laboratory methods of obtaining plant immunostimulatory preparations “Asgipan”, “Glabiloks” and “Sapanox”. Studied herbal preparations obtained by HPLC method from Aesculus hippocastanum, Glycyrrhiza glabra and Saponaria officinalis plant extracts. Scaling of HPLC process carried out in two ways: by increasing the amount of sorbent in preparative column and by varying the time of chromatography, gradient elution, the volume of eluent applied to the column per unit time. During the process of scaling carried out a permanent spectrometric control for the effectiveness of purification and quality of plant preparations “Asgipan”, “Glabiloks” and “Sapanox” was carried out. Studies have revealed that the change of eluent volume from 0.8 ml/min to 5 ml/min and the column type allow to increase the yield of the final product more than 12 times, due to the effective separation of the original plant increased in 6 times used for purification of “Asgipan”, “Glabiloks” and “Sapanox”. http://dx.doi.org/10.1016/j.jbiotec.2015.06.325 Evaluation of the biologic activity of some quaternary ammonium salts against non-pathogenic and pathogenic microorganisms Georgiana Horincar 1 , Ioana Otilia Ghinea 2 , Bianca Furdui 2 , Andreea Veronica Dediu 1 , Gabriela Bahrim 1 , Rodica Mihaela Dinica 2,∗ 1 Department of Food Science, Food Engineering and Applied Biotechnology, “Dunarea de Jos” University, Galati, Romania 2 Department of Chemistry, Physics and Environment, “Dunarea de Jos” University, Galati, Romania
E-mail address: [email protected] (R.M. Dinica). We present herein the antimicrobial activity on non-pathogenic and pathogenic microorganisms of some heterocyclic compounds, synthesized by us using non-conventional “green” methods, which are one of the most used classes of disinfectants, with wide application in hospital environments, water treatment, textiles, paint and food industry. The study on toxicity of heterocyclic compounds was performed by determining the biological activity on non-pathogenic and pathogenic microorganisms through the diffusion method and by studying the cell survival assay using 3-(4,5-dimethylthiazol2-yl)-2,5-diphenyltetrazolium bromide (MTT). MTT measures the mitochondrial function, being widely used to measure the cytotoxic potential of a chemical compound. The antimicrobial activity was strongest against Staphylococcus sp., Streptococcus sp., and E. coli, and the lowest was against Candida albicans. Some compounds, whose Staphylococcus and Streptococcus MIC are very small (≤250 mg/mL), may be of therapeutic interest. Acknowledgements: This work was supported by a grant of the Romanian National Authority for Scientific Research, CNCSUEFISCDI project number PN-II-ID-PCE-2011-3-0226. http://dx.doi.org/10.1016/j.jbiotec.2015.06.326
The susceptibility of chicken embryo to endotoxin solution eliminated by carbonized material Kuanysh T. Tastambek 1,∗ , Nuraly S. Akimbekov 1 , Ilya Digel 2 , Azhar A. Zhubanova 1 , Irina S. Savitskaya 1 , Gulzhamal Zh Abdieva 1 , Perizat S. Ualyeva 1 1 Al-Farabi Kazkah National University, Almaty, Kazakhstan 2 Aachen University of Applied Sciences, Julich, Germany
E-mail address: [email protected] (K.T. Tastambek). Bacterial endotoxin (LPS) removal from aqueous solutions is a challenging bioengineering task because of its ubiquity and high physical and chemical stability. Nowadays, adsorption methods of LPS removal are regarded as most advantageous with many different adsorbent materials being developed. In this work, low-cost nanostructured carbonized material (rice husks-CRH) designed by us in accordance for LPS-adsorption. The purpose of the present investigation was to determine the susceptibility of chicken embryos to an injection of endotoxin and CRH-treated endotoxin in vivo during the period of development. Endotoxin extracted from cell walls of E. coli by LPS extraction kid was injected intravenously into fertile eggs, of varying ages. It has been observed that the susceptibility (the development of provisional organs, pyrogenicity, blood analysis and lethal dose (LD50)) of chicken embryos to endotoxin decreased daily as the embryo matured from day 10 to hatching, while CRH-treated LPS had showed weak effect. These data show that the toxin effects of LPS solution was eliminated by CRH, according to the increased resistance of embryos. We consider the obtained results on the basis of novel advanced and economically efficient material CRH are very promising as the increasing number of biotechnological and biomedical applications supposed to be endotoxin-free, as well as still persisting problems with purification of complex mixtures. http://dx.doi.org/10.1016/j.jbiotec.2015.06.327