03882ntm a2200397 i 4500001001400000003000700014005001700021008004100038020002200079039011800101040002300219090002600242100003500268245013000303264003400433264001200467300007900479336002100558336002100579337002500600337002300625338002300648338003000671347002400701500005600725502009300781504004000874520203500914610007102949650004503020650001103065952012003076952013103196999002503327999013203352vtls000102809KUKTEM20251117113359.0180118s2017 my da f a m 000 0 eng d aTHE0000981(Local) 9a201905241626bnazirahc201802081544dfateehac201802021152dfateehac201801231143dfateehay201801181553zfateeha aUMPbengcUMPerda aFKKSA .N87 2017 r Bc.0 aNursyahira Mohd Rafi,eauthor.10aEffect of freeze and thaw cycle and incubation period on the solubilisation of inclusion body protein /cNursyahira Mohd Rafi 1aKuantan, Pahang :bUMP,c2017 4c© 2017 axiii, 70 pages :billustrations (some color), charts ;c30 cm. +e1 CD-ROM atext2rdacontent atext2rdacontent aunmediated2rdamedia acomputer2rdamedia avolume2rdacarrier acomputer disc2rdacarrier atext filebPDF2rda aFaculty of Chemical & Natural Resources Engineering aProject Paper (Bachelors of Chemical Engineering) -- Universiti Malaysia Pahang – 2017 aIncludes bibliographical references3 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.20aFaculty of Chemical & Natural Resources EngineeringxDissertations 0aUniversities and CollegesxDissertations 0aTheses 00104071a10000b10000d2019-09-04l0oFKKSA .N87 2017 r Bc.p0000122138r2019-09-04 00:00:00t1w2019-09-04yPSM 00104071a10000b10000d2019-09-04l0oCD 11149 | FKKSA .N87 2017 r Bc.p0000122139r2019-09-04 00:00:00t1w2019-09-04yPSM aVIRTUA40c7673d7679 aVTLSSORT0080*0200*0400*0900*1000*2450*2640*2641*3000*3360*3361*3370*3371*3380*3381*3470*5000*5020*5040*5200*6100*6500*6501*9992