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Proteomic analysis of Pseudomonas putida KT2440 during polyhydroxyalkanoates synthesis
Abstracts / New Biotechnology 33S (2016) S1–S213
solution at various salt concentrations and at 310 K. Sulfated modified levan structures are less compact than levan structures because of sulfate groups in the structures. Self-diffusion coefficients are calculated to investigate sulfate groups affect as decrease in movement of the structures. Diffusion coefficients decreased with increasing salt concentration for both unmodified and sulfated levan structures. http://dx.doi.org/10.1016/j.nbt.2016.06.1242
P17-2 Proteomic analysis of Pseudomonas putida KT2440 during polyhydroxyalkanoates synthesis ˙ Justyna Mozejko-Ciesielska University of Warmia and Mazury in Olsztyn, Poland As a result of rapid development associating with technological progress a demand for the biomaterials with unique properties is increasing. Among the biodegradable polymers polyhydroxyalkanotes (PHAs) are the most known. These bacterial polyesters are accumulated as discrete granules and used as a storage material for carbon and reducing equivalents. Generally, PHAs are synthesised when a carbon source is present in excess and an essential nutrient such as nitrogen, phosphate or oxygen is available in limited concentrations. Especially, medium-chain-length polyhydroxyalkanoates (mcl-PHAs) – a type of polyhydroxyalkanoates, have gained much interest in research on biopolymers because of their ease of chemical modification. Despite the highly satisfactory properties of PHAs, their production on a large scale is currently limited mainly due to high production costs. The biosynthesis process could be improved by a better understanding of how the metabolic networks that are responsible for mcl-PHAs synthesis respond to culture limiting conditions and the type of substrate. Therefore, the main goal of this study is to determine the influence of the nitrogen on the global changes in the protein expression profile during mcl-PHAs synthesis by Pseudomonas putida KT2440. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) have been used to investigate how substrates (sodium gluconate and oleic acid) and culture conditions (non-limiting and limiting concentrations of ammonium) influence on protein expression profiles. The obtained results enabled to identify proteins that are involved in the regulatory mechanisms of PHAs synthesis. The study is a good starting point to gain insight into the PHAs accumulation machinery.
S151
P17-4 Changes in the physico-chemical properties of bacterial cellulose depending on the synthesis time and Gluconacteobacter xylinus strain Radosław Drozd ∗ , Karol Fijałkowski, Rafał Rakoczy, Maciej ˙ ˛ Konopacki, Artur Frackowiak, Anna Zywicka West Pomeranian University of Technology in Szczecin, Poland The aim of this study was to analyze the impact of the synthesis time on the physico-chemical properties of bacterial cellulose (BC) produced by different strains of Gluconacetobacter xylinus. Cellulose samples were obtained from four reference strains of G. xylinus cultivated for 28 days in the stationary conditions using HerstinSchramm medium. The samples were analyzed using ATR-FTIR and XRD techniques. Furthermore, the ATR-FTIR spectra of BC were analyzed by means of the two-dimensional correlation (2D correlation) analysis. The ATR-FTIR 2D correlation spectra of the region from 1200 cm−1 to 800 cm−1 indicated the presence of the strain specific cross bands. The intensity of the observed changes in the 2D spectra indicated a significant effect of the culturing time on the molecular structure of BC. The significant decrease in the Ialfa crystalline phase within the cultivation time was observed for all of the analyzed BC samples. The rate of change in the structure of BC was found to be strain depeneded. Similarly, the XRD analysis indicated the decrease in the crystallites size occurring within the progressive changes in the proportion of crystalline phases of the BC obtained from cultures lasting for more than 10 days. Summarizing, the results obtained in the present study showed the differences in the physico-chemical properties of BC depending on the synthesis time and the bacterial strain used for its production. Acknowledgement: This study was supported by the National Centre for Research and Development in Poland (Grant No. LIDER/011/221/L-5/13/NCBR/2014). http://dx.doi.org/10.1016/j.nbt.2016.06.1245
P17-5 Sulfated Halomonas levan as a heparin-mimetic bioactive glycan Merve Erginer 1,∗ , Binnaz Coskunkan 2 , Tunc Morova 1 , Deniz Rende 3 , Seyda Bucak 2 , Nihat Baysal 3 , Rahmi Ozisik 3 , Mehmet S. Eroglu 1 , Mehmet Agirbasli 3 , Ebru Toksoy Öner 1 1
Marmara University, Turkey Yeditepe University, Turkey 3 Rensselaer Polytechnic Institute, United States 2
http://dx.doi.org/10.1016/j.nbt.2016.06.1243
P17-3 Withdrawn
http://dx.doi.org/10.1016/j.nbt.2016.06.1244
Highly sulfated glycosaminoglycan, heparin has been used as an anticoagulant and antithrombotic drug for more than 70 years. However, use of heparin has limitations such as contamination risk and production difficulties due to its animal origin and chemical inhomogeneity, therefore need for heparin mimetic compounds for therapeutic uses increased. These compounds are either chemically synthesized or, natural or chemically modified glycans having sulfate groups and exhibiting important biological properties including antioxidant, immunomodulating, antiherpetic, antitumor, antiviral, antithrombotic and anticoagulant activities. Considering the lack of reports about use of levan derivatives as heparin mimetic compounds, levan from Halomonas smyrnensis AAD6T chemically modified with various levels of sulfation. Biological activity tests such as biocompatibility, anticancer and anticoagulant activity (PT, APTT, TT and Heptest) were performed in vitro. Sulfated levan samples showed anticoagulation activity via
solution at various salt concentrations and at 310 K. Sulfated modified levan structures are less compact than levan structures because of sulfate groups in the structures. Self-diffusion coefficients are calculated to investigate sulfate groups affect as decrease in movement of the structures. Diffusion coefficients decreased with increasing salt concentration for both unmodified and sulfated levan structures. http://dx.doi.org/10.1016/j.nbt.2016.06.1242
P17-2 Proteomic analysis of Pseudomonas putida KT2440 during polyhydroxyalkanoates synthesis ˙ Justyna Mozejko-Ciesielska University of Warmia and Mazury in Olsztyn, Poland As a result of rapid development associating with technological progress a demand for the biomaterials with unique properties is increasing. Among the biodegradable polymers polyhydroxyalkanotes (PHAs) are the most known. These bacterial polyesters are accumulated as discrete granules and used as a storage material for carbon and reducing equivalents. Generally, PHAs are synthesised when a carbon source is present in excess and an essential nutrient such as nitrogen, phosphate or oxygen is available in limited concentrations. Especially, medium-chain-length polyhydroxyalkanoates (mcl-PHAs) – a type of polyhydroxyalkanoates, have gained much interest in research on biopolymers because of their ease of chemical modification. Despite the highly satisfactory properties of PHAs, their production on a large scale is currently limited mainly due to high production costs. The biosynthesis process could be improved by a better understanding of how the metabolic networks that are responsible for mcl-PHAs synthesis respond to culture limiting conditions and the type of substrate. Therefore, the main goal of this study is to determine the influence of the nitrogen on the global changes in the protein expression profile during mcl-PHAs synthesis by Pseudomonas putida KT2440. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) have been used to investigate how substrates (sodium gluconate and oleic acid) and culture conditions (non-limiting and limiting concentrations of ammonium) influence on protein expression profiles. The obtained results enabled to identify proteins that are involved in the regulatory mechanisms of PHAs synthesis. The study is a good starting point to gain insight into the PHAs accumulation machinery.
S151
P17-4 Changes in the physico-chemical properties of bacterial cellulose depending on the synthesis time and Gluconacteobacter xylinus strain Radosław Drozd ∗ , Karol Fijałkowski, Rafał Rakoczy, Maciej ˙ ˛ Konopacki, Artur Frackowiak, Anna Zywicka West Pomeranian University of Technology in Szczecin, Poland The aim of this study was to analyze the impact of the synthesis time on the physico-chemical properties of bacterial cellulose (BC) produced by different strains of Gluconacetobacter xylinus. Cellulose samples were obtained from four reference strains of G. xylinus cultivated for 28 days in the stationary conditions using HerstinSchramm medium. The samples were analyzed using ATR-FTIR and XRD techniques. Furthermore, the ATR-FTIR spectra of BC were analyzed by means of the two-dimensional correlation (2D correlation) analysis. The ATR-FTIR 2D correlation spectra of the region from 1200 cm−1 to 800 cm−1 indicated the presence of the strain specific cross bands. The intensity of the observed changes in the 2D spectra indicated a significant effect of the culturing time on the molecular structure of BC. The significant decrease in the Ialfa crystalline phase within the cultivation time was observed for all of the analyzed BC samples. The rate of change in the structure of BC was found to be strain depeneded. Similarly, the XRD analysis indicated the decrease in the crystallites size occurring within the progressive changes in the proportion of crystalline phases of the BC obtained from cultures lasting for more than 10 days. Summarizing, the results obtained in the present study showed the differences in the physico-chemical properties of BC depending on the synthesis time and the bacterial strain used for its production. Acknowledgement: This study was supported by the National Centre for Research and Development in Poland (Grant No. LIDER/011/221/L-5/13/NCBR/2014). http://dx.doi.org/10.1016/j.nbt.2016.06.1245
P17-5 Sulfated Halomonas levan as a heparin-mimetic bioactive glycan Merve Erginer 1,∗ , Binnaz Coskunkan 2 , Tunc Morova 1 , Deniz Rende 3 , Seyda Bucak 2 , Nihat Baysal 3 , Rahmi Ozisik 3 , Mehmet S. Eroglu 1 , Mehmet Agirbasli 3 , Ebru Toksoy Öner 1 1
Marmara University, Turkey Yeditepe University, Turkey 3 Rensselaer Polytechnic Institute, United States 2
http://dx.doi.org/10.1016/j.nbt.2016.06.1243
P17-3 Withdrawn
http://dx.doi.org/10.1016/j.nbt.2016.06.1244
Highly sulfated glycosaminoglycan, heparin has been used as an anticoagulant and antithrombotic drug for more than 70 years. However, use of heparin has limitations such as contamination risk and production difficulties due to its animal origin and chemical inhomogeneity, therefore need for heparin mimetic compounds for therapeutic uses increased. These compounds are either chemically synthesized or, natural or chemically modified glycans having sulfate groups and exhibiting important biological properties including antioxidant, immunomodulating, antiherpetic, antitumor, antiviral, antithrombotic and anticoagulant activities. Considering the lack of reports about use of levan derivatives as heparin mimetic compounds, levan from Halomonas smyrnensis AAD6T chemically modified with various levels of sulfation. Biological activity tests such as biocompatibility, anticancer and anticoagulant activity (PT, APTT, TT and Heptest) were performed in vitro. Sulfated levan samples showed anticoagulation activity via