Synthesis of poly(3-hydroxybutyrate) using enzymes Azospirillum brasilense in vitro
Introduction:Polyhydroxyalkaniates (PHAS) are bacterial polymers that are formed as naturally occurring storage polyesters by a wide range of microorganisms usually under unbalanced growth conditions. Mechanical properties of PHAS make them suitable replacements for petrochemically produced bulk pla...
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| Format: | Article |
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University of Isfahan
2017-06-01
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| Series: | Biological Journal of Microorganism |
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| Online Access: | http://bjm.ui.ac.ir/article_21567_28e6a3ea41dfb353be5c1d9bb4b8083c.pdf |
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| author | Soheila Abbasi Giti Emtiazi Rasoul Roghanian |
| author_facet | Soheila Abbasi Giti Emtiazi Rasoul Roghanian |
| author_sort | Soheila Abbasi |
| collection | DOAJ |
| description | Introduction:Polyhydroxyalkaniates (PHAS) are bacterial polymers that are formed as naturally occurring storage polyesters by a wide range of microorganisms usually under unbalanced growth conditions. Mechanical properties of PHAS make them suitable replacements for petrochemically produced bulk plastics (polyethylene, polypropylene etc.), but in contrast to these commodity plastics PHA are completely degradable to carbon dioxide and water through natural microbiological mineralization. PHAS can be produced by biotechnological processes under controlled conditions. Polyhydroxybotyric acid (PHB) was the first of thePHAS discovered and is the most abundant polyester found in bacteria. Materials and methods: First, a rich bacterial suspension prepared from poly(3-hydroxybutyrate), then the cells were broken down and ensure the absence of live cells.The cell free enzymes were added to M9 medium.Then the sample turbidity was measured at a wavelength of 600 nm and incubated at 30°C and centrifiuged at 120rpm. Next, the amount of PHB as well as solution turbidity were measured and compared with control. The physical and chemical analyzes of extracted PHB samples such as UV absorbtion, FTIR, HPLC and HNMR assesed and compared with standard sample. Results: In twelve days this experiment was conducted, from the third day, the amount of polyhydroxybutyrate was measurable and the highest polyhydroxybutyrate was gained on the tenth day. The level of PHB was constant. Moreover, the level of glucose was decreased gradually till the tenth day. By Physico-chemical analysis, production of polyhydroxybutyrate was approved. Discussion and conclusion: In this experiment, the reaction efficiency was 37.5 percent. Probably the rest of glucose could be changed to other intermediate compounds by other enzymes in the sample.Therefore, by purification of the enzyme, usage of specific substrate and optimization of conditions outside the cell could help the production of polyhydroxybutyrate continually. Production is very cost-effective, by this method. |
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| language | English |
| last_indexed | 2024-12-16T17:03:28Z |
| publishDate | 2017-06-01 |
| publisher | University of Isfahan |
| record_format | Article |
| series | Biological Journal of Microorganism |
| spelling | doaj.art-29ebf63b65eb4f2ca5a9c98bf82963222022-12-21T22:23:39ZengUniversity of IsfahanBiological Journal of Microorganism2322-51732322-51812017-06-0162212714221567Synthesis of poly(3-hydroxybutyrate) using enzymes Azospirillum brasilense in vitroSoheila Abbasi0Giti Emtiazi1Rasoul Roghanian2M.Sc. of Microbiology, University of Isfahan, IranProfessor of Microbiology, University of Isfahan, IranProfessor of Microbiology, University of Isfahan, IranIntroduction:Polyhydroxyalkaniates (PHAS) are bacterial polymers that are formed as naturally occurring storage polyesters by a wide range of microorganisms usually under unbalanced growth conditions. Mechanical properties of PHAS make them suitable replacements for petrochemically produced bulk plastics (polyethylene, polypropylene etc.), but in contrast to these commodity plastics PHA are completely degradable to carbon dioxide and water through natural microbiological mineralization. PHAS can be produced by biotechnological processes under controlled conditions. Polyhydroxybotyric acid (PHB) was the first of thePHAS discovered and is the most abundant polyester found in bacteria. Materials and methods: First, a rich bacterial suspension prepared from poly(3-hydroxybutyrate), then the cells were broken down and ensure the absence of live cells.The cell free enzymes were added to M9 medium.Then the sample turbidity was measured at a wavelength of 600 nm and incubated at 30°C and centrifiuged at 120rpm. Next, the amount of PHB as well as solution turbidity were measured and compared with control. The physical and chemical analyzes of extracted PHB samples such as UV absorbtion, FTIR, HPLC and HNMR assesed and compared with standard sample. Results: In twelve days this experiment was conducted, from the third day, the amount of polyhydroxybutyrate was measurable and the highest polyhydroxybutyrate was gained on the tenth day. The level of PHB was constant. Moreover, the level of glucose was decreased gradually till the tenth day. By Physico-chemical analysis, production of polyhydroxybutyrate was approved. Discussion and conclusion: In this experiment, the reaction efficiency was 37.5 percent. Probably the rest of glucose could be changed to other intermediate compounds by other enzymes in the sample.Therefore, by purification of the enzyme, usage of specific substrate and optimization of conditions outside the cell could help the production of polyhydroxybutyrate continually. Production is very cost-effective, by this method.http://bjm.ui.ac.ir/article_21567_28e6a3ea41dfb353be5c1d9bb4b8083c.pdfPHBPolyesterAzospirillum brasilenseHPLCNMRFTIR |
| spellingShingle | Soheila Abbasi Giti Emtiazi Rasoul Roghanian Synthesis of poly(3-hydroxybutyrate) using enzymes Azospirillum brasilense in vitro Biological Journal of Microorganism PHB Polyester Azospirillum brasilense HPLC NMR FTIR |
| title | Synthesis of poly(3-hydroxybutyrate) using enzymes Azospirillum brasilense in vitro |
| title_full | Synthesis of poly(3-hydroxybutyrate) using enzymes Azospirillum brasilense in vitro |
| title_fullStr | Synthesis of poly(3-hydroxybutyrate) using enzymes Azospirillum brasilense in vitro |
| title_full_unstemmed | Synthesis of poly(3-hydroxybutyrate) using enzymes Azospirillum brasilense in vitro |
| title_short | Synthesis of poly(3-hydroxybutyrate) using enzymes Azospirillum brasilense in vitro |
| title_sort | synthesis of poly 3 hydroxybutyrate using enzymes azospirillum brasilense in vitro |
| topic | PHB Polyester Azospirillum brasilense HPLC NMR FTIR |
| url | http://bjm.ui.ac.ir/article_21567_28e6a3ea41dfb353be5c1d9bb4b8083c.pdf |
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