Purification and characterization of papain enzyme from papaya / Norashikin Zulkaple

Project paper (Bachelor of Chemical Engineering (Biotechnology)) -- Universiti Malaysia Pahang – 2012

Bibliography : p. 40-42

Purification and characterization of papain enzyme isolated from unripe Carica papaya fruit is proposed by using expanded bed adsorption chromatography. Papain also known as cysteine protease is a type of industrial enzyme. The high request of this enzyme has encourage research involving the purification of papain, in the most efficient was by reducing cost and time span and increasing the purity and concentration and recovery ratio of the product. The expanded bed adsorption process was carried out using Fastline 10 column and Amberlite XAD7HP as the adsorbent. The latex of unripe papaya was diluted in buffer solution. Before starting the expanded bed adsorption operation, there are certain parameters were determined in order to achieve optimum conditions for the expanded bed adsorption process. The binding buffer and elution buffer that is suitable for each stage was determined in percentage of ethanol. After that, the expansion characteristic of adsorbent Amberlite XAD7HP was determined. Then, the dynamic binding capacity of the adsorption stage was studied. Accordingly to the parameters that have been determined, the expanded bed adsorption process was operated based on the condition that has been determined. From the batch adsorption in the binding stage and elution stage, it was found that 10% ethanol was suitable for binding stage and 90% ethanol was suitable for elution stage. The maximum adsorption capacity obtained is 13.14 μg and the velocity when H/H0 is two has the value of 2.6 x 10-6 mm/s. The dissociation constant,Kd in the Langmuir equation is 6.97 μg/ml. Profile for expanded bed adsorption process is plotted and the three stage of expanded bed adsorption process can be seen where the stages include binding stage, washing stage and elution stage.