Extraction and Purification of Enzyme Actinidin from Kiwi Fruit (Actinidia chinencis)
الباحث الأول:
Rasha Adnan Kazem
الباحثين الآخرين:
Adnan Wahhsb Habeeb
المجلة:
Plant Archives
تاريخ النشر:
1 مايو، 2020
مختصر البحث:
The aim of this study is to extract, purify and estimate molecule weight of the actinidin enzyme from kiwi fruit. The extraction process is carried out using a buffer consists of 6% sodium chloride + 2% boric acid in which it results in having 39 un…
The aim of this study is to extract, purify and estimate molecule weight of the actinidin enzyme from kiwi fruit. The extraction process is carried out using a buffer consists of 6% sodium chloride + 2% boric acid in which it results in having 39 units/mg quality activity for the actinidin. After that, the process of purification is started by concentrated the crude, that is resulted from the extraction process, by saline gradient using ammonium phosphate with a saturation ratio varying from (20-80)%. At 60% saturation, the highest enzymatic activity of 48.7 units/ml is achieved. Then, the dialysis was carried out using potassium phosphate and the specific activity resulted from this step is 128.3 units/mg protein. The purification process was completed using ion exchange chromatography column DEAE - Sephadex A-50 and a quality activity at this step is 192 units / mg protein. Then, gel filtration with a column Sephadex G-75 is used in which it results in having an enzymatic activity of 20.4 units /ml and a specific activity of 226.6 units/mg protein at 5.81 number of purification and 11.33% enzymatic yield. Using electrophoresis, the purification process reveals that there is one protein bundle which is one of the signs to the purity of the enzyme. Moreover, the molecule weight of actinidin enzyme which is 245000 Dalton has been determined.
