000 03607ntm a2200373 i 4500
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008 180118s2017 my da f a m 000 0 eng d
020 _aTHE0000981(Local)
039 9 _a201905241626
_bnazirah
_c201802081544
_dfateeha
_c201802021152
_dfateeha
_c201801231143
_dfateeha
_y201801181553
_zfateeha
040 _aUMP
_beng
_cUMP
_erda
090 _aFKKSA .N87 2017 r Bc.
100 0 _aNursyahira Mohd Rafi,
_eauthor.
245 1 0 _aEffect of freeze and thaw cycle and incubation period on the solubilisation of inclusion body protein /
_cNursyahira Mohd Rafi
264 1 _aKuantan, Pahang :
_bUMP,
_c2017
264 4 _c© 2017
300 _axiii, 70 pages :
_billustrations (some color), charts ;
_c30 cm. +
_e1 CD-ROM
336 _atext
_2rdacontent
336 _atext
_2rdacontent
337 _aunmediated
_2rdamedia
337 _acomputer
_2rdamedia
338 _avolume
_2rdacarrier
338 _acomputer disc
_2rdacarrier
347 _atext file
_bPDF
_2rda
500 _aFaculty of Chemical & Natural Resources Engineering
502 _aProject Paper (Bachelors of Chemical Engineering) -- Universiti Malaysia Pahang – 2017
504 _aIncludes bibliographical references
520 3 _aOverexpression of recombinant protein in bacteria result in the formation of inactive protein. These inactive proteins associate forming insoluble protein aggregates which is referring to inclusion bodies (IBs). Generally, IBs are pure and the aggregated protein inside it has native-like secondary structure which is a bioactive protein. To recover the insoluble and active protein is a major problem encountered. Solubilisation does play a crucial role by unfolded the protein and thus help it to refold properly so that functional bioactive protein can be recovered. Example of mild solubilisation method using low concentration of urea and combine with freeze and thaw method has been proven to increase the efficiency of the recovering of bioactive protein. For freeze and thaw process there are factors that affect the overall process which are freezing incubation period and number of process cycle. Incubation period affect the process by determining the amount of stress needed to be applied so that unfolding process can occur. Number of cycle does affect the protein stability in terms of the occurrence of protein degradation probability. Thus the objective of this research is to investigate effect of freeze and thaw cycle an incubation period on the solubilisation of IBs. For incubation period the experiment was conducted between 1 to 4 days whereas for freeze and thaw cycle experiment conducted between cycle 1 to 4. Moreover, the performance of these two parameters were analysed using native-polyacrylamide gel electrophoresis (n-PAGE) to determine the functional for enhance green fluorescent protein (EGFP) amount and Bradford assay to determine the total protein amount. In this study, incubation period did affected the performance of solubilisation rate in which the IBs being solubilised and then proceed for refolding process and has been proven achieved active form of EGFP. For number of process cycle, it did not affect the solubilisation rate on determining the amount of functional EGFP recovered.
610 2 0 _aFaculty of Chemical & Natural Resources Engineering
_xDissertations
650 0 _aUniversities and Colleges
_xDissertations
650 0 _aTheses
999 _aVIRTUA40
_c7673
_d7679
999 _aVTLSSORT0080*0200*0400*0900*1000*2450*2640*2641*3000*3360*3361*3370*3371*3380*3381*3470*5000*5020*5040*5200*6100*6500*6501*9992