Protein Engineering _ Siti Julaiha

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    Jck)ici`hzh lsked sk}sl lcuhihhk ld}h mhk zhpduhk lcskdlhk ihudkj ihudkj

    dkmdqdms+

    Lc}dlh jck `clcpfh mckjhk `hdl/ }s`sb ld}h `cplci`hkj mhk icidodld askjud thkj

    `hdl+ ]h~d `doh uc`shb jck }skjjho ) `hblhk ucjick lcedo mhpd uc`shb jck }skjjho ickjhohid

    lclhehshk/ mh~h} `cpcacl mckjhk hmhkth eheh} mhk ~ckthld}/ `hblhk lcih}dhk+

    Jck hmhohb us`skd} ~clcpfh mhpd MKH+

    MKH hmhohb `osc ~pdk} mhpd uc}dh~ dkmdqshodu gpjhkdui+ Icps~hlhk mh}h`huc dkagpihud

    ldidh `cuhp thkj ici`hzh uc} dku}pslud ockjlh~ sk}sl ici`sh} ucish ~pg}cdk thkj hlhk

    md`s}sblhk }s`sb+ Uc}dh~ jck `cpdud ucphkjlhdhk dku}pslud }cp}ck}s/ `dhuhkth ickjlgmclhk

    ~pg}cdk }cp}ck}s+ Hjhp uco mh~h} ici`sh} ~pg}cdk/ dkagpihud mhpd jck mduhodk/ `hudu mckjhk

    `hudu/ mhpd MKH lc mhohi hosp `hps mhpd icuuckjcp PKH "iPKH$+ Lcismdhk iPKH `cpfhohkhk

    lcoshp kslocsu icksfs lc ud}g~ohuih/ mhk ~hmh `hjdhk uco thkj mduc`s} gpjhkco pd`gugi/

    iPKH ickjhphblhk ~cphld}hk huhi hidkg thkj }cpod~h} ickfhmd igoclso ~pg}cdk+

    MKH bhmdp uc`hjhd msh ~huhkj phk}hd ~hkfhkj/ icodkjlhp mhohi `ck}sl bcodlu

    jhkmh+ Uc}dh~ sk}hd }cpmdpd mhpd fs}hhk `ogl `hkjskhk ldidh thkj mduc`s} `huc+ Ucick}hph

    bhkth hmh ci~h} mhuhp ldidh thkj `cp`cmh mhohi MKH thd}s Hmckdk/ ]didk/ Ud}gudk/ mhk

    Jshkdk/ sps}hk `huc)`huc thkj }cpfhmd ickck}slhk dkagpihud thkj }cpucmdh+

    MKH `cphmh ~hmh dk}d/ h}hs kslocsu/ ihudkj)ihudkj mhpd }pdodskhk uco }s`sb+ Uc}dh~ uco

    ihksudh "mckjhk ~ckjceshodhk uco mhphb icphb/ thkj }dmhl icidodld dk}d$ `cpdud MKH thkj

    uhih+ Uc}dh~ uco icidodld =1 igoclso MKH sk}hd jhkmh+ Uc}dh~ igoclso }cpmdpd mhpd 3>)

    03>>>>>>> hudu }cpudi~hk ~hmh+ lpgigugi+

    MKH uc}dh~ lpgigugi ickjhkmskj `hkthl jck mhk ici`ck}hkj oshu/ }cpihusl kgk

    lgmdkj MKH thkj askjudkth `cosi mdlc}hbsd+

    H~h d}s ^pg}cdk7

    Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 8

    @DG]CLKGOGJD

    PCLHTHUH ^PG]CDK

    Ud}d Fsohdbh ' >4>134:=0>

    ^hueh Uhpfhkh]clkgogjd @dgicmdu

    Skdqcpud}hu Dkmgkcudh

    ^hueh Uhpfhkh ]clkgogjd @dgicmduSkdqcpud}hu Dkmgkcudh

    \Icd 0>8>Y

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    ^pg}cdk hmhohb ususkhk phk}hd ~godicp thkj }cp`ck}sl mhpd udk}cuh huhi hidkg huhi hidkg

    mhohi phkjlhdhk lgi~oclu `cp`hjhd fckdu dlh}hk mhk u}psl}sp ucbdkjjh }cp`ck}sl fckdu fckdu

    ~pg}cdk thkj `cp`cmh uh}s uhih ohdk+

    Mhohi }s`sb ihksudh/ ~pg}cdk }cp`ck}sl mhpd mhpd udk}cuh thkj }cpfhmd mhohi uco mdisohd mhpd

    udk}cuh phk}hd jhkmh MKH thkj }cpmh~h} mhohi kslocsu uco thkj ickjhohid ~pgucu udk}cuh}phkulpd~ud ickfhmd phk}hd }skjjho PKH mhk mhpd phk}hd PKH }cpuc`s} ickjhohid ~pgucu

    }phkulpd~ud mckjhk hmhkth pd`gugi ucbdkjjh }cp`ck}sl ~pgucu ~ci`ck}slhk phk}hd jhkmh

    mckjhk ~pgucu ~ci`ck}slhk agomdkj h}hs od~h}hk od~h}hk phk}hd ickfhmd phk}hd lgi~oclu mhohi

    u}psl}sp thkj `cp`cmh+

    PCLHTHUH ^PG]CDK

    U}psl}sp ~pg}cdk thkj `cp`cmh icktc`h`lhk }cpfhmdkth ~cp`cmhhk askjud fsjh ~hmh ~pg}cdk

    ~pg}cdk thkj }cp`ck}sl mhohi uco+

    Lhphl}cpdu}dl huhi hidkg uhkjh} md~cposlhk lc}dlh pclhthuh ~pg}cdk mdjskhlhk sk}sl icisohd

    ush}s ~pgtcl+ @dhuhkth ~pgtcl thkj `cp}sfshk icohlslhk ~cps`hbhk ~hmh ~pg}cdk thkj }cohb

    cludu h}hs hmh/ icisohd ohkjlhb hzhokth mckjhk ~ckck}shk huhi hidkg thkj mdjskhlhk

    sk}sl icked~}hlhk is}hud+ Bho dkd uhkjh} ~ck}dkj lhpckh lgk}pd`sud huhi hidkg }cpbhmh~

    askjud ~pg}cdk icps`hb udah} mhk lhphl}cpdu}dl ~pg}cdk/ ucbdkjjh ~ckjc}hbshk h}hs hkhoduh

    }cpbhmh~ lhphl}cpdu}dl huhi hidkg bhpsu }cohb md~dldplhk+ Lc}dlc icpclhthuh ~pg}cdk sk}sl

    askjud mhk lhphl}cpdudl kgqco'`hps/ bho thkj }dmhl sisi khisk ickhpdl sk}sl mdohlslhk

    hmhohb ickjh}sp `c`cph~h is}hk ickjjskhlhk huhi hidkg thkj `cpqhpdhud mhk fsjh

    icpclhthuh is}hud ucehph iso}d~oc ~hmh uh}s ~pg}cdk+

    Pclhthuh ~pg}cdk hmhohb h~oodlhud ehi~sphk ickhpdl mhpd ~ckjc}hbshk 6

    ) @dgogjd Igoclsocp

    ) Hkhoduh U}psl}sp Ldidh pg}cdk

    ) Ih}cih}dlh

    ) Clgkgid

    ) Lgi~s}hud h}hs @dg Dkagpih}dlh

    ) @dgldidh

    sk}sl ici~pgucu ~ckjci`hkjhk ~pg}cdk ickfhmd ~pgmsl thkj `cpjskh mhk `cpbhpjh+

    Icps~hlhk mdud~odk dois thkj mdlh}cjgpdlhk ismh sudhkth/ mckjhk ~ckcod}dhk ~ckcod}dhk thkj

    Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 0

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    uhh} dkd mdohlslhk sk}sl icihbhid u}psl}sp dlh}hk ~pg}cdk/ pclgjkhud ~pg}cdk sk}sl ~pdkud~

    ~pdkud~ mcuhdk thkj mh~h} md}cph~lhk+

    UCFHPHB PCLHTHUH PG]CDK

    Pclhthuh ~pg}cdk mdisohd mhpd ~ckcishk ~ckcishk }cpbhmh~ ckvdi mhk askjud

    askjudkth/ thd}s ~hmh }hbsk

    8255 Hkucoic ^htck! Fchk)Aphkgdu ^cpugv mhpd ^cphkedu/ icohlslhk

    Dugohud mhpd lgi~ocl hidohud mhpd jcpidkh}dkj hpoct mhk ickhihlhkkth

    mdhu}huc

    U}hpeb usjhpu "iho}guc$

    8253 F+ Fhlg` @cpvcodku mhpd Uzcmdh

    Ickmciguk}phudlhk `hbzhqu}hpeb mh~h} md~cehblhk ucehph cadudck ickjjskhlhkclu}phlud iho}+

    Bho thkj mdlckho mckjhk khihLh}hodudu

    8251 ]bcgmgp Uebzhkk mhpd Fcpihk

    Ickjdugohud ~c~udk/ ckvdi ~cp}hih thkj mducmdhlhk mhpd fhpdkjhk bczhk+

    82:1 Zdoodhi Lbkc

    Ususkhk }dmhl }cph}sp 'Skgpjhkdvcm acpick acpick mddu}dohblhk ickfhmd Ck vdi thkjmd~c}dl mhpd `hbhuh tskhkd `cphuho mhpd Ck `cphpd dk mhk vtic `cphp}d ohqck'phjd ~hmhtchu}

    824: @sebkcp `cpuhsmhph `cphuho mhpd Fcpihk

    Ickjclu}phlud ucehph `c`hu uco uco Tchu}

    Joseguc c}bhkgo , EG0

    ]SFSHK PCLHTHUH PG]CDK

    ) Ickjci`hkjlhk ~pg}cdk sk}sl ucush}s thkj `cpjskh h}hs `cpbhpjh md `dmhkj dkmsu}pd h}hs

    lcucbh}hk +

    ) Ici`ck}sl ~pgmsl hps mhpd ush}hs ehi~sphk ~pg}cdk

    ) Icps`hb u}psl}sp h}hs udah} ~pg}cdk ucbdkjjh mh~h} mdbhudolhk ~cps`hbhk ~hmh ~pgmsl ~pg}cdk

    mckjhk }hpjc} u}psl}sp h}hs udah} thkj mddkjdklhk+

    ) Icps`hb ~pg}cdk ucbdkjjh mh~h} md}cpfcihblhk sk}sl ici`cpdlhk dkagpihud mhk h~odlhud ohdk+

    ) Ickjhkhoduh udah} ~pg}cdk thkj hmh sk}sl h~odlhud ~pgmsl ohdk

    Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 5

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    ) Ickdkjlh}lhk lcihi~shk mhk cadudckud ~ckjjskhhk ~pgmsl sk}sl ush}s ~pgmsl

    LCJDH]HK PCLHTHUH ^PG]CDK

    ^ckdkjlh}hk lh}hodudu mckjhk us`u}ph} hohid h}hs lgahl}gp

    ^ckdkjlh}hk lh}hodudu mckjhk us`u}ph} kgkkh}spho h}hs lgahl}gp

    ^ckdkjlh}hk lh}hodudu md kgkkh}spho ~cohps}

    ^ckdkjlh}hk odjhk ickjdlh}

    U][email protected]]HU ^PG]CDK

    ^ckdkjlh}hk }cpigu}h`dod}hu

    ^ckdkjlh}hk hl}dqd}hu ~hmh ~B ho}cpkh}da

    ^ckdkjlh}hk hl}dad}hu mhohi lclsh}hk dgk thkj `cp`cmh

    ^ckdkjlh}hk ~pg}cdk od~h}

    ^ckspskhk lcpck}hkhk }cpbhmh~ ~pg}cgodudu

    Ahpihlgldkc}dl

    CLU^PCUD ^PG]CDK

    ^ckdkjlh}hk }dkjlh} clu~pcud md bgu} kgkkh}spho

    ]hpjc} clu~pcud lc oglhud ucosohp thkj `cp`cmh

    ^ckhi`hbhk }hj sk}sl ickmc}clud clu~pcud ~pg}cdk

    ^cps`hbhk igmdadlhud ~gu}}phkuoh}dgkho

    Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc =

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    UDAH])UDAH] OHDK

    Ickhi`hblhk }hj sk}sl iciahudod}hud ~cispkdhk

    Ickjs`hb lceckmcpskjhk sk}sl ~godicpduhud

    Ickhi`hblhkk }hj sk}sl iciqdushoduhudlhk oglhoduhud

    Icpclhthuh ickjdlh} ud}su hogu}cpdl

    Ickjs`hb }d}dl dugcocl}pdl

    Ickspsklhk diskgjckdud}hu

    H ODLHUD PCLHTHUH ^PG]CDK

    ) ^ci`sh}hk ckvdi

    ) Ici~pgmslud fckdu g`h} g`h}hk

    ) Ickjbhudolhk qhludk hk}d`gmt }cp}ck}s

    @c`cph~h Egk}gb H~odlhud ohkjuskj ucehph lgicpudho uc`hjhd `cpdls} 6

    ) Ickjjslhkhk ~cphkehb'uehaagom ~pg}cdk `cpmhuhplhk mgihdk ~hmh ^pg}cdk H sk}slickjci`hkjlhk igoclso igocls uc~cp}d hk}d`gmt "Egi~hkt 6 Haad`gmt$

    ) Ickhi`hblhk huhi hidkg thkj }dmhl }cplgmclhk lh~hmh ~pg}cdk/ iciskjldklhk udk}cuh

    ~pg}cdk mckjhk mdqcpud}t `hbhk ldidh "Egi~hkt 6 Hi`pr$

    ) Ickjjskhlhk ~cphkehb ~pg}cdk `cpmhuhplhk ~cp~dkmhbhk sk}sl ickjci`hkjlhk igoclso

    igoclso uc~cp}d hk}d`gmt/ ici`sh} ~ckjjh`skjhk ~pg}cdk ~pg}cdk "Asud$ mhk pcec~}gp

    hjgkdu} "Egi~hkt6 @dgPcrdu$

    ) Ickjjskhlhk ~cphkehb ~pg}cdk `cpmhuhplhk oce}dk ]d~c E sk}sl ickjci`hkjlhk igoclso

    igoclso uc~cp}d hk}d`gmt/ }clkgogjd ~pg}cdk }pdicpduhud "Egi~hkt 6 @gpchk$+

    ) Icpclhthuh ~pg}chucu sk}sl ickspsklhk igoclso igoclso thkj md}hpjc}lhk "Egi~hkt6

    Eh}hotu}$+

    Ud}d Fsohdbh'^hueh Uhpfhkh ]clkgogjd @dgicmdu SD ^hjc 3

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    ) Ickjjskhlhk mgihdk Ad`pgkcl}dk sk}sl ickjci`hkjlhk igoclso igoclso uc~cp}d hk}d`gmt/

    icked~}hlhk ~pg}cdk ~pg}cdk msh askjud mckjhk mgihdk dlh}hk }hpjc} thkj }cpbs`skj

    }cpbhmh~ mgihdk ckvdih}dl "Egi~hkt6 Egi~gskm ]bcp~hcs}deu$+

    ) ^ckjci`hkjhk ~ckdkjlh}hk ic}gmc sk}sl bsihkduhud hk}d`gmt "Egi~hkt6 Lhog`dgu$

    ) Ickjjskhlhk ~cphkehb ~pg}cdk `cpmhuhplhk od~gehodk sk}sl ickjci`hkjlhk igoclso

    igoclso uc~cp}d hk}d`gmt "Egi~hkt6 dcpdu$

    ) Ickjjskhlhk ~cphkehb ~pg}cdk `cpmhuhplhk Lpdu}ho jhiih sk}sl ickjci`hkjlhk

    igoclso igoclso uc~cp}d hk}d`gmt "Egi~hkt6 Uedo$

    ) Ickjjskhlhk ~cphkehb ~pg}cdk `cpmhuhplhk Udu}cdk Lkg}u sk}sl ickjci`hkjlhk igoclso

    igoclso uc~cp}d hk}d`gmt "Egi~hkt6 Ucocegpc$

    ) Icpclhthuh askjud cacl}gp thkj mddkjdklhk }cpbhmh~ ~pg}cdk ~pg}cdk UID^ uc~cp}d hk}d`gmt

    "Egi~hkt 6 ]ps`dgk$+

    ) Ickjjskhlhk }clkgogjd ^MH sk}sl Icpclhthuh askjud cacl}gp thkj mddkjdklhk }cpbhmh~

    hk}d gmt mhk icked~}hlhk ~pg}cdk ~pg}cdk mgihdk kcjh}dqc mhk ~pg}cdk ~pg}cdk mckjhk

    ~ckdkjlh}hk udah} ~pg~cp}dkth "Egi~hkt 6 Rckegp$+

    ) Phkehkjhk mhk lgku}pslud mhpd ~pg}cdk h}hs ckvdi `hps mckjhk }clkgogjd `hps'kgqco h}hs

    askjud askjud thkj