استخراج و غني سازي الكل دي هيدروژناز از جگر مرغ با بكار گيري ميكروحباب ها﴿ Aphron﴾

چكيده انگليسي :

Chicken Liver Alcohol Dehydrogenase Extraction and Enrichment using Microbubbles Aphrons Afsaneh Goortanian Nejad a goortanian@ce iut ac ir Date of Submisson 22 September 2013 Department of Chemical Engineering IsfahanUniversity of Technology Isfahan 84156 83111 Iran Degree M Sc Language Farsi Supervisors Kayghobad Shams k shams@cc iut ac ir Abstract Plant tissues and animal organs are the most important sources of enzymes in the growing field of enzyme biotechnology About 70 of the enzymes are extracted from plant tissues or exudates and animal organs Animal derived enzymes represent about 10 of the total enzyme market Alcohol dehydrogenases ADH are a group of dehydrogenase enzymes that occur in many organisms and facilitate the interconversion between alcohols and aldehydes or ketones It is believed that chicken liver is a good source of ADHs Therefore in the present study chicken liver was selected as an affordable accessible animal organ which is rich in a vast variety of enzymes including ADH and the goal of this study was extraction and enrichment of ADH from chicken liver Recovery and purification of biological products is a complex process that usually its production cost is relatively higher than the normal commodities and goods In recent years the production of new proteins in the biotechnology industry using simple and inexpensive separation methods has attracted much attention Conventional protein purification processes are multistep processes that involve a significant loss of enzyme activity One of the promising methods for enzyme separation and enrichment is foam fractionation It is believed that separation of proteins using foam would minimize the separation steps and would maintain satisfactory purification efficiency In this method generated bubbles in the solutions that contain surfactants and enzymes create a gas liquid interface that adsorb the active substances have shown a good potential for separation of enzymes with high efficiency This is because of the fact that aphrons would enhance the transfer of the enzyme to the aphron phase by increasing the gas liquid interface through reduced bubble size and as a result increased specific mass transfer area So in this research project micro bubbles were used for the extraction and enrichment of ADH enzyme In order to produce bubbles ultrasonic homogenizer at various speeds and in different time spans was used to agitate and blend the surfactant containing aqueous solutions Then a certain volume of bubbles was mixed with solution containing the enzyme mixture and enough contact time was spared for enzyme isolation To investigate the effects of mixing and surfactant on possible enzyme denaturation of the isolated enzymes electrophoresis was used Enzyme concentration was determined via light absorption of UV and Beer Lambert correlation was used to calculate the enzyme concentration The results showed that the type and concentration of the surfactants pH and the presence of salts had profound effects on enrichment At first experiments were performed using anionic and cationic surfactants and finally AOT was selected as the best surfactant The salts used for extraction were KCl and NaCl Results showed that in the presence of KCl and at a concentration of 6 mM AOT and at a pH value of 6 smaller bubbles were generated and the highest enrichment yield was obtained 238 9 Measured bubble sizes using light microscope were about 10 100 micrometer Experimental observations indicated that generation of nano sized bubbles in presence of KCl would be plausible however because of the unavailability of suitable measuring devices identification of nanobubbles was not explored In addition electrophoresis results confirmed the enrichment and indicated that the isolated Alcohol Dehydrogenases were not denatured Keywords Alcohol Dehydrogenase enrichment Aphron Electrophoresis PDF created with pdfFactory trial version www pdffactory com

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استخراج و غني سازي الكل دي هيدروژناز از جگر مرغ با بكار گيري ميكروحباب ها﴿ Aphron﴾

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