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008 131009t2013 my da f m 000 0 eng d
020 _aTHE0002246(Local)
039 9 _a201905241114
_bshamsul
_c201710061522
_daishah
_c201311220748
_dnabilah
_c201311201556
_dnabilah
_y201310091225
_znabilah
040 _aUMP
090 _aQP601 .Y84 2013 rs Thesis
100 1 _aYuhai, He
245 1 0 _aDirect recovery of papain from papaya juice using expanded bed adsorption chromatography /
_cHe Yuhai
260 _aKuantan, Pahang :
_bUMP,
_c2013
300 _axvii, 80 p. :
_bill. ;
_c30 cm. +
_e1 CD-ROM
502 _aThesis (Master of Engineering (Bioprocess Engineering)) -- Universiti Malaysia Pahang – 2013
504 _aBibliography : p. 67-77
520 3 _aPapain is an enzyme from Carica papaya which of great research interest in recent years due to its pharmaceutical and biological application. The adsorption of papain on AmberliteTM XAD7HP adsorbent with the mechanism of reverse phase liquid chromatography (RPLC) was investigated in this study. The equilibrium adsorption isotherm, binding buffer and elution buffer conditions were studied in a batch system. 15% ethanol was obtained as the optimum binding buffer with binding efficiency of 76.53% and 40% ethanol was confirmed as the optimum elution buffer with elution efficiency of 84.09%. At the optimum conditions, the adsorption capacity of AmberliteTM XAD7HP for papain was found to be 4.5 mg papain/g adsorbent. The equilibrium isotherm was fitted to Freundlich isotherm which has an n value of 1.3927 and the K of the adsorption of papain onto the AmberliteTM was 267.67 (mg/g)(ml/mg)n, R2 is 0.9746. Papain in the concentrated extract was purified 4.3-folds and 55% purity corresponding to around 15% of the total papain in the crude juice. More than 84% of the adsorbed papain was eluted using 40% ethanol as the elution buffer. The efficiency of purifying papain from papaya juice using the method developed above was around 65.5%. A purified papain with 65% yield and 50% purity was recovered. A direct recovery of papain enzyme from unclarified Carica papaya juice was developed successfully using a reversed phase expanded bed adsorption chromatography (RP-EBAC). The dynamic binding capacity for the RP-EBAC at 10% breakthrough of 810.52 μg papain/g adsorbent was achieved at a linear flow velocity of 936 cm/h, bed expansion degree of 2 and feedstock viscosity of 2.68 mPa·s. Papain purity of 58.39% and purification factor of 4.96 was obtained by one-step elution in RP-EBAC. Two-step elution was employed to enhance the purity of papain in RP-EBAC which gives higher papain purity of 74.98% and higher purification factor of 7.04. Absolute ethanol (100%) was successfully used in the Clean-in-place (CIP) for cleaning of the AmberliteTM XAD7HP adsorbent. After the first CIP about 15% of total protein was still retained on the adsorbent and about 3% of total protein still retained on the adsorbent after second CIP. It shows that there is no strong cumulative effect of contaminants happens on the adsorbent. This work shows a great potential of using RP-EBAC to purify papain from unclarified Carica papaya juice.
650 0 _aPapain
650 0 _aAdsorption
650 0 _aPapaya juice
856 4 0 _uhttp://ecollib.ump.edu.my/25640/
_zLibrary access only
999 _aVIRTUA40
_c4036
_d4042
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