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$)RTSTEK

PUSPIPTEX

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tsBN 978-979-98404-9-3

00il0Gotrf.r.nco on !ndustd.l Enzth. .nd Elot.chnologtf

ProceedingEnzyme Technology

Serpong, TangorangSeminar, 3 August 2010j

t

'1Ftra .Flz!'ilaLrE!

'l', I,-

Deputy ofTechnolow toa Agroindustry and BioteclnolAgencyfor the Assessm€nt d|d Application of Tecrnobgy

cdt r.E n hdsti.l E|lm rnd g,ti.dnoldvlots r6{n'F;'w Trlmloft fe EsFndlv hdr6a.1a|d 2OlO

ISBN 97&979-9U04-9-3

PROCEEDING:CONFERENCE ON INDUSTRIAL ENZYMES AND

BToTECHNOLOGY (CIEB 2010)

Enryme Technology

for Eco-Friendly Industry

Orgrrlr.d byl

CeEter for T€chnolos/ of Blo hduttryGoverrrn€nt Agency for Acsgamert .trd Appllcrtlor of TechDology

itr cooParadon with

R6e.rcb Ccller for saleDc{3 rDd Techlolos/MlDlrtry of Relc|rch rrd Tecbtrolog/

rnd

IteutlcherAtrdemilchcr Au*rNchdlcnat (GerDrtr AcadeEicErcbrtrge S€rvice)

T.nSer| g' 2010

cdtr@d l"d,nti.l Elrm.,n €ldldtn ro.v lotl to10l'Fid T.dftldv tr FaFnadrY I ||d'.I tu 2rlo

ISBN 978-979-9U04-9-3

PROCEEDING:COI\IFIRENCE ON INDUSTRIAL ENZYMES AI\ID

BIOTECENOLOGY (CIEB 2010)

Enzyme Technology

for Eco-Friendly Industry

Tangerang, Brnten 03 August 2010

Edibd by:

Budiasih WahlrntadSuyanio Pawiroharsono

Sise.a Setyrhadils Hclia$iTrismilah

@ All nght r€served

Publish€d bY:

Center for Technolog/ of BlolndwtryGovernment Ag€ncy for Ass6sment rnd Applicrdon of Technologl

(BPPT)BPPT bullding 2, 15'floor' Jl' MH Thrmrln 8, Jrkrrtr 10340

Telp. 02r-31G9513. FrI. 021-31G9510

20t0

cdt r.* d hd6ti: Fnrh. nd Blft('jn ldt loti 2otol'Ellzft T-Iialr,e fr FGFndd! bdr6/. I A'|d 20lo

Itrtroduction

Enzym€s ar! bio.atalysts gbich scceleratc a certah Oio) cheDicil r€aclioa a[d foud in €vcry

cell of atl living beiDgs, fifi|r sirele singlc ccllubr or8ldsrDs to highly conplex flrlticellularorgadisrds. Enzyn€s phy 6n ir4ona role in our live, not only in our oEn hctlbolism, but also ino|h€i proccss€s susrlhing our dsily activiti€s. Thcy are us€d lo sssist tll€ productiotr of th€ food *ccat aDd contribule to the carE of ou. healh by lroviding tb€rspeutic aSenls or scBitive a.d sp.ci6cdirgdo€tic t .{s. Thus ir tss hid conerci.l rd indusr.ful Yahrc.

Enzytrs also play atr irDponrot tolc in 'gftcn indstry'. lDrtrtd of using mor€ hszardous

cbcmicals, €trzyrmtic proccss is c rhore envitonrnentslly fti€ndly choic€. For cxerqle, ifftead ofusing cblorinc for tbc bldhing p.ocess€s, ptot ax. is a 'gt€toet' choice for detcrgrd as wells pulp

asd paF hdlrstri€s.

In additio& prodll;ls oore cnviotrmtrttlly fticrdly proc6s.d 'It

oow in higi6 dcmt s, dr€ to

tigbt r €nvironrFntrt rrgulstions, elpccialy in thc dcvclop€d colmni€s. C6s€s involviry rcjeclio[s ofhdon.siso erpo.ted prodlcts by thc envirorlDedlalists bccsus. thc rnalufacturing prcc.sses were

corBidcrcd not cnvironEeotdly ftieodly should ocvcr hle occufirc4 hsd ih€ dcciliod nakcrs appli.dsi.ic-t r l0r,i rrrd h5d tbc iDdustislisb hsd rmtE lwarEn€ss oo eDvironDnld issrB EDrymd'tbercfcr€, would b€ otre ofth€ answ€ts.

The drta tom oor rnattet rr3clrcb c4osutianl indicstod that b 20(X thc vahE of.DzyDe deInald

h Asia Pa.ific was 640 trlillions US dollst, whcr€ 387 millions US dolats is lssociated withindustial coryrns sd 253 US do[rrs wilh spccirlty enzyms us€d in tle pbsmac€utic,L te*"tchaDd di.gtro*ic field6. Atdougb lbc Ftrctr k lcrs pEdictablc, IDdo.si.tr dorncstic d.mrn& s.dr ro

iDcrEasc quitc slcadily betwcar 2001-2005, r€achiry nol€ thsn 3500 tons id totsl iD 2O0t Tbc biShctl

denad scems to b€ ftom the dc&rgpnt industry, cotrsisti4 on av€tagi 15% of lotal- Th€ ftsts wet€

u!.d iD oths industrial s€ctorB, iocludina l€ath6 p.ocassin& clnlc fcad' tatil€3' F p ad pqF .slxcl ,s food and pbedcluticals. Unfortuaatcly, thc whole anru.n is s1i[ colqlctcly i4orted

Co6i&ring lte ev€t incrcssiog 4Plicatioo of €'lzlD.s in hdl5tri€s' wodd$i& as wcll 3s

don€srically, ii is ooly rcasonrblc lhsl rls€8rcb.fi i! lhe ficld of biocsttlysb I wel as dccisi{tn

drsters siut to lhiDl about this pmbl€t!" Itr lldoncsi4 wherr i[dultries arc not yct cloccty liokcd wilhtbe res€€Irh srd scadthic world, a€s€a&h prodlcts would al*rys b€ mercly rcsearch ptodualr ullessthc d.cisioo rmt.rs tssdt tbciruulbority in policy Ir|AkiDg, supPortiDg thc .pplicstion of &tmcsdc

rH.arcb Fodwrs in odion l hd$ai€s" Tb€rdor€, in or&t lo bring rbe indusEirlists' cdrynecscrchcrs and dccision natcr closcr togelher, wc orgaoiscd ltc Coofccocc oo bdrstritl EszfneBiotcchmlory 2010 (C!EB 2OlO), which wrs hcld iD Targrtalg otr 3 - 6 August 2010' wftt ritlcEtrzyDe T.chlolo$/ for f,.!-Frlctdly IDdutfy.

T|llcr|rg,3 Angutt 2{10

Dir.ctor ofCent€r for BioiMustrial Techtrolog/

Wilooo B&suli

Conf.r.M. m rndunrEl t.sm. ..d Bbr..Jlnolon lcrFs 2or0l'Em Tajlrollp fn E*Fnddlt lnd!6/ . I Auo< 2010

Co[teDa

P18€

Introductios lti

List ofcoolenl iv

IndBtrinl Etrztme Prcduction ia B&t.ti.: C,.netics rnd Sttlitr lopmvdEnt

Friedhcta Meinhardt vlll

BioconvcrsioD: From BioE ss lo Cbcnicsls atd Plod.rts

Klnus-Dicter Vorlop, Frsn! Scbucbrdt, Hcinz SticbDothc, Tboms Willte |!

C\lIrlnt Stste ofEtrzync R€scarch fu Indon€sia

Massy Tbenawidjajr Suhiiooo tl!

P4l kolation aDd Id.ltification ofTb€dmphilic C€Uulasc-Ptodrcing Bactdia fiom M€rap

Volcm Craler ln Wcst Sum3t€ra

Jantssri. YusniwEti, Aswaldi Anr,sr, B€Dtri Ismul Huda I

PA hohtion OfThernostsblc -Amyhse From Th€mophilic Ba.l€.is Of Soiai Hot

Spring SouIb East Sulawcsi

Sspto R.ahrrjo, Andi Noor Kholidhssysrifin, L.M. Rtntdben, Prima Endang' S 7

P43 Scllaritrg lor cellutasc Foducilg b€ctcria lrom Talifugu n bripes fish

Diatr Andtiad, Bambabg Pras€tya, Don H€e Pa* l7

P{4 Scr! itinS ofDt€rmstrblc Cellulrs€ tltd Xihnase Producing Microo.gtntutB

Prima Endaag Susilovui: S€plo RrharF; ArdiaDsysb, Rthrmwad Rthirq samiMrreeti, Ahnd Zsctri A

P45 Selcctiotr ofTr}?cin litc proteasc p.oducing lactic rid tuclcria atrd cbaErlcriation ofthc eozyne

Dysh Wutatrrari, Budisih Wahlutsri 30

P46 Clh$iclcrizarbn of Allrlo-themrybilic XylaDss. Prodrrciry Bsdeda boLt€d from

Rokrn Hulu, RiaD

Shafs Noor, Ytineri*arni; Trismilth t7

P4? ScrE iting of EDzyne Aclivity amng Fuogal and BeLrial Isolatcs as Biocoatol

Agent ofcbbroot Dis.asr (PlasDodiophora brossicac) on Ctbbegc (Br&ssicr

olcraccae).

Bahsruddin, Tutit Kuswintnti aod Cusmiaty 17

P-08 Xylmligo6scchtridd fuu.tion ftorn Corncob Xylan uting Sttptotnyc6co6t rictolls 451-3 Enzy.nc.

Adr Mctyaoditril, Titi Csdra Suttni2, EDma Msutta Moko3 52

P-09 Pote irsl ofAlgd BionDss for Biodicsel Prcducing Thtougb EtrzynrticTralr.st rification olMicmalgal Filty Acids

JoLo Sulirtyo, Ritr D*i Rrbayu aDd Yati Sudtryiti Sockr a

cdnffi d rhd6tn.l E.:wR .nd srd.cnn ld' lcrai 20101 'Erffi T6htlo'v fd feFdadlv lldr6l- 3 AId 2010

P-10 OprinrlizatioD ofcbiliDsse Foducrio, by B.cilus 5P Cl5 and chtr"act€dzrlion oftbe

Ruby S.tisctsD, Budirsih Wshyutrttri

P-l I Optimizstion ofAcration rnd Cooubstntc addition in Xilosc Bioconvcrsion htoXiliiol by Cadi& Ttopicllis

I*smi Anborssri, SutyaDi, FraDsislo Elr tludaFni

P-12 Optiniution OfNa-Alginrtc AItd C!cl2 CoFttrtrstion For lromobiliz.d Cc[ CmdidsTropicalis Itr Xylitol Ptoduction

Surysd, Irtsmi Amb€.srri, Dyts HisrDysnti

P-13 Tbc rfr€ct ofT.ichl,orfon m Cbolircst r.sc Activity of Difietltrt OrgrDs ofBhEMusscl, Mytilus ed is.

L Ysoio aDd P--D. HtDlcn

P-14 lhc Efrcct ofYoghutl Th.mpy on Pmfilc and Activig ofDigestive Ellz''EG (PmE"se,

Lipasc rd suc1..se) Expc.d by Fonnrfdehy& or Rttrus iov€fgiq$ Jejuum

ftr RlhErtul tryly, Aulami'an, ChstrifMthdi

P- I 5 Thc lrtrp.ovement of L-srabino6e isomcrase local Gcob.cillus stearolhctttbphilusAcidic tolerarce for Tagalose hdulbirlizstion by Sit€-Dfucct€d Mutag€ocsh of Glu-269

Dcwi Fitriad, Lita Trirsbr atrd Budi S€Isono 125

P-16 Cyclodcrt in Gtyco6yl Trrnsfcnse Synthaic CeD€ ofBs.illua 3p Ar-5r:OveExpr€ssion iD Eschcrichia c.li and Ctaracletizriion ofthc producl

Catur Rirni, Alhnsd Muchlisin, Casilawsti, Debbie Sotie Rehodngrum 130

P-l? Clodng phytase g@ ofKlieh's€Ia prFumnia asil itrio c.coli bl2l (de3) .trd

chslsctctizarion of tbc cozyn€

Adi Magn! Patriadi Nuhriru.angsa, Z3€oal Bacbruddio, Sajidttr, Ali Wibowo l35

P-18 Cloning OfTbernosbbl€ Dna Polymqsee C€tre Fron A Thqmophilic Br€vibaciuus

Sp. IBolal€d Fmm Sikidary Cmlsr, DcrS Plalar, Ccdtnl Java

Lucir Dhiaditr Witsssri, kfrtr Dwija Prijrobsda! Jafs Widrda 14

P-19 CbanclcrizationofBioclcctochcmicrlElecEod€of Phstrctochaetechrysodporiurn

Flavin Domain Celobiosc Debydrogentsc Reconbinrnt (PcCDtI), Substrale Atrtrity,Optinum pH ad Storsgc Stability

D$riari, Slcftm FERRT Koji SODE t5,,

P-20 Atplicarion ofEnzym LiDtcd lrDtDuno6oftcnt Assay (ELISA) for hrlv Dete.tion ofPLRV, P\D( ad PVY on Atlantic Varicty Potalo s€€d G€n€mtion 0 (C0) and

Genedtiotr 2 (G2)

Tutit KuswiMnti. A. Mtsdia*sti, Adc Rosnstra dsn BahaDddin 155

P-21 Th€ Us€ of Mol€culsr Tccbdque in Cath€cin Corcentrsiion Diagnocis of Ur.otttgrndir Roxb

Istioo Fcri6', JaDsari, Irfan Suliatrryrh, Austis4 Hamda Fsuzg dan Yusniwati 150

l0l

I ConEspo.dios.'nhor, itrinofciit'@Fboo.cot'

PA2 D.tectiotr of Aedd r.S/pti ard Ac. ilbopictB Vins h Deryuc Fever EDdanic iltPrdug City using Rcvcts€ Trsmciptala Polyrretasc Chain Rcaction Mclhod (RT-rcR)

Hssmiwati, D.bclmi, Nurhayati

P-23 Por. ial for Protein Dcposition in Diffcrr Btoile. Ccnotypcs Dependiry on Age

161

Sarnadi t16

Pt4 l.ovaslatia Cooteft Produ.lion By Mut tr Offupergilus Tcnclrs Using Solid

FcfirEntation Procrss Atrd Liquid Fcllicnt'liotr Prccess

Triatr' Rias Dewi, Adc SuDiatdi, Susi Putwasih ltlP:25 Effect OfFemcDtation With Addition Slrrbio Probiotica OD Nutsient Conldt OfRice

Straw (Oryza Sativa L)

A& sumiardi, Suli Hctinswari 190

P:26 S)orE Mclhods to Detlct lh. Ecotoxic of Agroch€micrlsotr Mictobidl Activity io SoiI

FcrbbatrTirdaotr 198

Pt1 Tbe Uptrke ofcadmium loos md lls Influance on Thc Cro*'tb and hoduciiotr ofSomc S€codary Mcbbolitcs in Shoot CultuEs of Solfllum mcloogcna

TjicKok 2lls

P-28 Ctdlicsl Caqo6itioo OfEss€otitl Oil Ofsouth Surnatran Pittosporufi FenugiEunL AiI.

Ad€k ZntDrud AdnsD, Z{iouddin, Asmtdi Zls

ORCAl{IZINC COMMITTEE 216

P-27The UPtrke ofCrdmlum lonr rnd 16 Inlh€nce on The Grcwtl rnd

Producdor ofsome Sccondrry Mctrbolltec In Sboot Culturcr ofso'4T |,'tmdorScng

TJb t(o|('

Fedty of Biot cholo$t, Mrrt oatica aDd Scicacc Dqcrb'otUniv€tlity of surshoyr' susb6ya' IDdmcoia

A'STRACT

Tbc sim of this $|rdy *!3 to ctaniDc lhc tltpoBc givco by shoot cuhr€ of &'l@ild't tttlo'Be"a

Woo rpt ti|tt ofclddiun iotr& Fm ltc rledls of ttfu rac.rch' it could bc sarad thst s&oor odturE

of M@ui,rclotae,/gwc.cablc to grow in lhc o.dis cont itrirg 'lGl50IM c.dnium ios, wilh tbc

gm*th i[dcx gr! t r tb.o tbrcc; wtilc rpplirtim of bigh.i coocant dioa of lhd ioos will r6ult itr

tbc d.arh of th q t!c.. Thc crrltutt! w.t! eblc to r@vc 2O,2"J4.+/. ot b i@8 iom ncdi!

cootlitritrg tlGl6o pM of lb. iotrs tld eloddrd 6.n b thcL ttoosls. Arpli'c.tim of 120 tMcldmioro ioos lo thc E dis cau.d r dacc.rc b lolrl fte phyroEolr c6t nt (50.2y.) of ltcculotr!3.

Iftyiordtj Ctfuium iot, lhoot c1|hrc, So/otttt tlerota€ta, ph)'lct tok.

INTRODUC'TTON

CldmiuE k ! qbgs&3l-d9494 $/iO ltc syEbol Cd sDd alomic number 48, occ|trs wid! ZilE oNts.

Crdoius i3 uscd l&gely ir b.lcrics sDd Distqqd!, fot .rtrylc itr pb$i! prodEts, !!d foosn io

caurc canc€r. C!fuiunr is rbo r potaotid covimo8ntal bazrd Hu''|m cxposut6 to cnvirol|codcadmiud rI! Firrldly lhc tld t of tbc hrDiDg of f6sil firb sd dltridFl w!rt6. Howcvcr, th.rlbv. b.co dbl€ ins@6 of loxicity I tbc |lsolt of lonSi'r.tm cxpdn€ lo ctdmiud iD

contaDriDrod food aDd *d€r. ID thc dacrd€ follosing ll&dd-Wa!!, Jqatrcs. ttritritr8 op.f,.lid3cootrmiDrrcd thc Jin ! Rivcr with crdmium !!d Esc6 of olhcf ioxic m.l8ls. Ar a conlcqu.!.qcadroium accumulrtrd in lhc ricc ctoDd grc*iry sloog tic ri\atb.Dlt &woslrcaD of sa nic. Th.local egriqdturd csEn|nitit'3 cmsmiDg thc coot dilrtrd ri:c dcv.lopcd ltei_ioi diteasc .Dd r@llhdo@liti:s, itrcftdiDg plalgulr! tDd gluc@ri& Cldnium is oe of six substaoc€ b.DDcd by thc

EurtDa€! Unioob Rrrtrdion m lt zrtdous Subct|rc€s ditEcrivc, wticb h.o3 carciloSlat m

coqot f3 (AmoiD,2m8).

CrddiuD sDd zinc ara h th. !!E grorp on tbc Fildic trbla, conrdl 6c lac coNm oxidrtioo

srdc (+2), ud *hao ionizcd.tr d6t tbc satr size. DrE l,o tb.3c aimibriti.', cadmium c.n n?leziDc iD may biologicd syst@, io Frricular, rys&43 that codtai! son.f [g.Dds $ch .. adfinCad|nium c.tr biod rp l,o tao tim ootl rtugt tbr zioc b c.rtti! biologfu.l tystd, aod i8

mtoriously difFq l to rcmvc. h ldditioo, c{dlsiui! cro rtple maqnesium sDd lglgigE in octlsiD

biotogicd iFt6E3, rlthoogh th€sc rtpltcctrots u! rtrc (AnooiE, 2mt).

Corft .ponding &!hor, tjic-ko&@Fhoo.cm

208

cdl@ d ftnd, !m r{'d!&!tcEr toiol'kEt&Y ntF^atti'db bddl. I Adtolo

Phrt ccll culturcs, ss wcll.s irs hrrcr phDI crn give ccrtair rEsponscs on thc lrlsarce of ccrtainmct&l ions in thc medium. Cltlnrcs of Chlorc a wlgaris could lelnov€ 75% of 8 UM coppfi ion6

fion lhc tn€dium (HemsD, 195). Hairy rool culturB of An &racia n/Jti.ona coluld rennve 62.4./0 oflhc coppcr ioDs tom initial \Eluc of 330 FM copp€r ioss (Zilrurndova 1995). CopE r tolEa calbs(-,ulfircs oI Nicoaiorra tab@utt rcge&rated pl"trls thal we.E able to gro$r iD thc prcacnc. of 100 FMcoppef ior|s (cori ct 31., 1998).

Tbc Fodu.aioD of scrquit rp@id ph)loslcxitri itr ccll silpctrsiotr qrlti.es of Lhttra starnonnortbave b€cn itrdrc.d by mctal ions, iDcluding cad&ium ions Clhrcfsll ad Wtiteh€a4 1989).

Th€ rEsulls of prcccdhg rcs.archs denoost-ald fiat i roducing rEtal iobs, iDcMitrg crfuium roos,into pletrt cell culNres could inducc, incrcas€, or iohibit thc goductiotr of sorra sccondary mctabolitcsin that cullur€s.

E'OERIMENTAL

l,tateriab

Thc shoot cuhur.s oflo/@irr tttclongern wer. cttltivsl|.,l od rmdificd Mu.sshige ltrd StooS neditwith lhe addition of 2 mg.L ' bcnrylsdenin€. Cultures were rnaintainrd h the ligbl using philipsTL40w a! 25oC. Subcuhuring of tb€ siock-shoots was pcffofmcd ev€ry 4 wecks (IDdn)€lto ct at.,1996).

h o.der to study dre accumuletiotr of c{dmium ioos and irs infbeoce on tbc prodlrcti@ of som€sccondary melabolites, shoot culturEs were tm$fcrr€d to a s€rics of20 mL mcdia coDtdning diff€notcorrc.ntratiotr of cadmium iotrs sDd cultivatd for 4 w€€ks udcr tb€ saoc conditiod prior toharvcstilg. For each csdmiuh iotrs conccntsatior! 15 botd€s of +l-l.j gram fr€sb weight of shootrcach wcrc us€d. Affer incubation thc cultu.cs rr€r€ hrrvestc4 weidc4 h b dried at 400C, and

Gtowth shdy

Thc gowlh of $hool culurcs wrs cxFessed h grodrr t der (CD, dcfm.d as the ratio of final fiEsbw.i8nt rd iaitid filsh weiSbl of.ach culrre. Th. CI r,!s msurod for cach bottle.

Analysb ofcodttin iont

Cadiium ions in the biom.ss wc& dctermined by using tCp-AES aftd apFoprial,, digestion of 0.1mg powd6 using 2.0 ml coocedtrsted HNq fouow€d by dihxion with dirtijld wrtei uD io 10.0 ,nl ordilutld fimher as r€quircd {Cb€n, t993}. qrbd6catjon ws! pcrfonDcd by me{surio; dle crtrlssimitrtcosiry ofcrdmirtu rr ib maximum *lvclcngth. The coocentration ofcsdmium iotrs \lr.s calculstcdfton the lio€a. rcgrrssim cquaiion ofsiandard-

Rf,SIJLTS AND DISCUSSION

Grow,h sbrdy

Th€ Esult6 of otB.rvatioa of tbc gm*th rrr€ crpr6s.d itr growtb ind.x (CD and th€ d€crrrsc ofsucrGe cooccrtsetioD in mcdia bcfore add affer hsrvesting w€r€ sbown oa Tsblc t , FigutE I aDd 2.

2(B

co.{Ru d hdu*nd Eh:ft ..d sdxn.oldv lcrFs 20101 '|ff. Tdhmlle fc E@FndnN bd's'. I Aud l0l0

Tabfe L The Srowth index ot the shmt cultut€s of &)/orrtn nelongena aged 4 we€ks and the

decr€ase ofsucros€ concentration in media before and aft€r harvesting

ConcetrFationofcd"

(Fl'f)

ReplicationCroMhind€x

D€slasc of sucaose

('/"1

40 l5 1.801{.84 64.50+0.74

80 t5 6.32+1.58 10.2o10.61

t20 l5 6.80+2.00 28.42+0.58

t60 4_58+1.84 24.51+0.55

The gto*'th i[dcx wt3 cxpc*scd s av6rga + d€vialion staodar( cbnngp in $E osc cooccntsation as

o/o avemce + devirlioo statdard.

9.(D

6.tb

7.00

6m

5.CD

4.00

3.00

2.m

1_q)

0.0080 1m

coldffi..ac'n+ (!M)

Figue | . Graph of gmwth iDdex vanatjon of So/d,uD nclongena .shoot culturs aged 4 week with lhe increasitrg concentratioD ofcd'

210

conr.r.nc. on rndunrul tnrw€ .nd Bbt.chnolo4 lclEs ?0101 'Idh. tdbnolor for trccFnddlv !'dudd'. I Auon l0l0

Figure 2. Graph of lbe decrcrx€ of sucaos€ corccntrstio! i! meditwith variatioo ofc? conccoEarion

Table I a Figur€ I $owed thalth€ sioot cultues of Solan/n ,Elongenz \|ere sble to grow in the

mcdfu contrining cadmium iotrs coaccntmtion up to 160 IM with lbe gD*tb index grratcr thao thr€e.Thc growlh lal! ioq€aled witb lbc ioc'rrosiry cnnc.otratioD of cadmium ioos, r@h€d ils maximur! sribe conc€aEation of rpproximdely 120 pM atd dccrcascd anErw!.ds. It was sactr ftom Table I,Figures I ad 2, rhat tbe growth rate was not olways invefcly proportiooal to rhe dcc-rease of sucnoceconcentralion ir ocdi& lt wes possibly due to tbe diff€Encc itr stEorption of nu[ie|nt otb.r tbm3UCr6€ by lbc Cultrcs.

Anal ys is oI p ht tos,e ro I ( s )

Tbc rrsults of tbc a$\sis of stcroidsi Slyco6idca(c) usilrg .nisald€iydc.Hzs0. .s s sprsyidg rlsgedtw€rt sbowD on table 2,

Trble 2. Alrlysis ofs{croidsl glyco6idds) using titr hycr ct ornatography Cn C}

Concetr!!lionofcd'-(rM)

St€roidal glycoside(s)

Resull Rf

zt0 0.63 (ete.n), 0.71(ycllow),0.63 (violel)

80 0.63 (rrEen), 0.71(ye[ow],0.81 (violet)

1200.63 (g€€tr), 0.71(yellow). 0.83 (violei)

1600.63 (seen), 0.71(yellow). 0.81 (viol€r)

+ indical€s that s substance being id€ntified is existing, Rf is ret6rdation factor

211.

8o 120

Cmtd-dcd'+(rM)

cdlslc s r^donn.l €'lm .nd Bi.n(hnotdv lclEB 20101 'En4 Trhhddv ra EeFndilv indus". I aud ml0

Tablc 2 showcd ihat pmdrcing steoidal glycoeid{6) in th€ir cclls, lhe shoot cullurts of So/aruttnclongata gave cettain s8 calo|r affer being spr.yed wilb adsald€hyde-H1so..

Accunulation ofcadntun iots and determino.ion of,he totallree sterob conten in bionoss

The r€sults of tbc ob6crvatioo of accumulatiotr of cadmiun iotrs and lolrl frEc st€rolx cont nt rD

btonals of &)lanun tnelongerra cdlure3 aaed 4 weeks wcr! demodt"aled otr Trble 3, Figur.3 ard 4.

Tsble 3. Thc accumulatioD ofc.dmium ioos rnd the total ftEe sl€rols c.nlcd in biomass ofsorar@rrrelangeru cultur€s aged 4 w€eks

Concent_atioD ofcadrnium ioDs aod lotal frE€ stcmls *ls expressed in,4/g dry weight ofbiornsss

Concctruali,otofcd"(nM)

ReplicationCorceo!-ation ofcd' h bionrss(ltziP\

Conccntrtiotr oftotd fie€ sterols

lqc,/c.:)40 t5 3.6+02 568,!9880 l5 52.0+1.4 450+75t20 l5 101.2+4.6 421+g)160 t5 135.9+2.1 324+87

150.0

125.0

1oo 0

75.0

50.0

25.0

00a{) 80 120 t6t)

c...-d .a cdt+ (rM) h ..d.

Figurc 3. cr+h ofcoratnrion ofcd:+ in bionasswith variation ofC<P concedration h m.dia

272

cdtdr@ d rndurn.lE'l2tm rnd Bdchnol@ lclls 20101"E M. Tdhmlln fs lilFndilv I'dud".3 AUE( 2010

C.!€t t ld .. tdl ltt

Figuc 4. Graph of conccotntion of lotal F€e sterols withvari.tiotr ofcd cotrcenEation in m€dia

Tabfc 3 ard Figur€ 3 showod that drc shoot cultu.Es of Solut rn melongena were sblc !o grow ro therncdia contlining crdmium iotrs corccntralioo W to t60 pM and ablc to rEmove 20.2-34.4%crdmium ioos ftom media .oatsittidg 4G160 IM of th€ ior|s rtld accumlatcd tbcm in their biomass.Applicatiotr of l20IM csfuium ioos to tbe n€dia caus€d a deql€se in totsl fi€€ pb)1ost6Dls conr4ar(,4.7/6, of rtts culturls. MorEover, Tsble 3 ard Figut 4 also dermr8Fat€d tbat appticrtion ofcadmium ions concenlration caused a d€crcare in total free stcmls cortant of the shoot culfurEs ofSolat tnn ,Elongetn. lt wB tbc higlcr the cadmjum ions cotrcctru"tioA th. higb6 the d.cr.a!€ inlotal ft€c stcaols contcnt.

CONCLUSIONS

ll could b€ concluded ftom the.!s€arEh, thst:

l. Tbe shoot orltuls of blamt nelongam wae ablc to gow h tbc rncdia conieining cldmitlmions conccntalioo up to 160 FM aod ablc to .qDove 20.2-A.4o/o of cl/,mfum ioos from medi.containing,lGl60 pM ofthc ions snd accumulat€d lhcm in thcir biorDr.ss. Applicatiotr of 120 lrM€dmiuE ions to lbc m.di! c-auscd s d€cretsc in rorsl fi€. phytoctcrols conr€d (502yt of lhc

2. Inrroducidg crdrnium ions into shmt cultu.cs of Solannn nelongem corid iDiibit the Foductionof tot l ftc phttod.rok in rh.si plrlt cc culnt|ls.

AC|<NOWLEIICEMENTS

I would likc to express my gratitude to all those wbo glvc rne thc pGsibility to conplck thjs tEs€srch.I waol to lhsnt ih€ Di.€ctor.lE CenerBl of Higher Edrcation R€Fblik lndorcsia for firun inS rE toaccor4list this res€arcb and my iD.stitutioL th€ Unive.sity of Sur 6ya, for givin8 rne faciliti€s toperfonn lhis r€r€dch.

8oo

6q)

5@

3@

2@

0

213

8o !20 160

cF.rL.dclo+(.M)r.d

co.Lreme on rndunnal tnrh. and Bioi.rhnolo& lcraB 20101 "EiMc Tehroldv fs Eclhmdr lndNF . I Arasr 1ol0

Rf,FENENCES

ul Anonin! 2008. Ilttp:i/ep.wikioedia.orq.iwiki/( admium (Accessed 7 Juni 2008).

[2] Hennan EB, 1995. Reced AdvaDces in Plad Tissue Cuhrc III. Regcner.tion andMicropropagatio[ Techoques, Systems aod Media. Agrite.h C.onsultads, Inc., Sbrub Oak.

tll Zihutrdo\a M., Maletcmva Y & Varcl T, 1995. Utilizstion of Plant Cell CultuEs forBiorcm€diation. ln: 43rd Annual ConSgr€s oD M€dicinsl Pl6nt Researcb, SecondaryProducls Physiologically Active Compourds, p. 43.

[4] Cori P, Scbiff S, SanLdr€a G & B€trtrici A, 1998. Req|otrs€ of In Vitro Cultur6 oflicoriazdtabocrn L. to Coppe Str*s ad Selectiotr of Plsnts &om Cu Tolcrsd Callw. PlaDr Ccll Ti$. Org.C\ L 53:l6l-169.

[5] ThrEfr[ DR & [/hitchesd IM, 1989. The u!€ of metal ions to induc€ th€ foroarion of sccoMyproducts in planl ticsuc cultu!. Io: RobiDs RJ & Rhodcs MJC (eds) M"tripulaliq SecondsryMetabolism iD Culturc. Camb.idge UniveFity PrEss, Canbridgc.

[6] Indl!]nio G, Utrmi W & Syabrani A 1996. Agaw onaniensb "Ir€l & Nowell: 1, ?r:tro cuuues3nd tle prcdrctioo ofphytGtcroids. Ill: Bajaj YPS (ed) Medicimt a.d tuonatic.

[7] Chen F, 1993. Nitricr'pcmxide digestion of biological tisuca. And],ticat Scrvic€ LaboraroriesTrace Meilk Metodolo6/.

2\4

confer.nceon lidusrtu Endde.nd Biot.chnoloR lclEB 20101"Enzvft Tahnoloqv lor EccFf,endlv hdustry"- 3 aueusl2010

ORGANIZIN(; COMMITTEI.,CIF]II2O1O

Advtuory Bo.rd: Dr.Ir. Liesryani wijayand- Deputy chairmanfor Agroindustrial Techoology andBiotechnology

Stccring Cornnittee: Dr. Wilono Basuki - Direator of PTB, BPPTAchrnadin Luthfi M., M.Eng. Head ofBiocaralyst DivisionDr. Siswa Setyahadi PfB, BPPrTe34 FfB, BPPT

Or$[bing Codmittee:CAslrpe.soo Dr. Astutiati NuhasanahSecElsriat Ika Rachrnatul l,aily, M.Si.

Ir. Rita RachmawariIr. Noor Farida Hasanah

Trersuer Dr. Niknik NurhayatiPublication Dr. Dadang Suhendar

Ir. Deden R. Wahaq M.Si.

Scientific Committret Dr. S. Budiasih WahluntariProf. Dr. Suyanto PawiroharsonoDr. Is HeliantiDr. Siswa Setyahadik. Trismilah, M.Si.

Confcrence: Dr. S. Budiasih WahyunrariDjamil, M.SiPuguh WidodoAyi MufriSupriyadiErnu Widyasri, S.Si.

WorkshoFr Dr. Is Helianti- Mol€culrr Biolog/ Wor*shop Dr. Vanny Nadta

Maria Ulfa, S.Si.Lina Mulianti

- prorain purincarioo worksbop *. 3;.i:ol?lilf*"""Achmadin Luthfi M., M.Eng.Dr. Dadang SuhendrrElizaberh, M.Si.

216