The uptake of cadmium ios and its influence on the growth and production of some secondary metabolites in shoot cultures of solanum melongena - Ubaya Repository Tjie Kok CIEB 2010
tsBN 978-979-98404-9-3
00il0
Gotrf.r.nco on !ndustd.l Enzth. .nd Elot.chnologtf
Proceeding
Enzyme Technology
Serpong, Tangorang
Seminar, 3 August 2010
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RTSTEK
PUSPIPTEX
Deputy ofTechnolow toa Agroindustry and Bioteclnol
Agencyfor the Assessm€nt d|d Application of Tecrnobgy
Suppoftd by
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 hduttry
Goverrrn€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
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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 Wahlrntad
Suyanio Pawiroharsono
Sise.a Setyrhadi
ls Hclia$i
Trismilah
@
All nght r€served
Publish€d bY:
Center for Technolog/ of Blolndwtry
Government 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
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 flrlticellular
orgadisrds. Enzyn€s phy 6n ir4ona role in our live, not only in our oEn hctlbolism, but also in
o|h€i proccss€s susrlhing our dsily activiti€s. Thcy are us€d lo sssist tll€ productiotr of th€ food *c
cat aDd contribule to the carE of ou. healh by lroviding tb€rspeutic aSenls or scBitive a.d sp.ci6c
dirgdo€tic t .{s. Thus ir tss hid conerci.l rd indusr.ful Yahrc.
Enzym€s
Enzytrs also play atr irDponrot tolc in 'gftcn indstry'. lDrtrtd of using mor€ hszardous
c rhore envitonrnentslly fti€ndly choic€. For cxerqle, ifftead of
using cblorinc for tbc bldhing p.ocess€s, ptot ax. is a 'gt€toet' choice for detcrgrd as wells pulp
cbcmicals, €trzyrmtic proccss is
asd
paF
hdlrstri€s.
oow in higi6 dcmt s, dr€ to
In additio& prodll;ls oore cnviotrmtrttlly fticrdly proc6s.d
'It C6s€s involviry rcjeclio[s of
tigbt r €nvironrFntrt rrgulstions, elpccialy in thc dcvclop€d colmni€s.
hdon.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.d
si.ic-t r l0r,i rrrd h5d tbc iDdustislisb hsd rmtE lwarEn€ss oo eDvironDnld issrB EDrymd'
tbercfcr€, would b€ otre ofth€ answ€ts.
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 with
industial coryrns sd 253 US do[rrs wilh spccirlty enzyms us€d in tle pbsmac€utic,L te*"tch
aDd di.gtro*ic field6. Atdougb lbc Ftrctr k lcrs pEdictablc, IDdo.si.tr dorncstic d.mrn& s.dr ro
The drta
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 .s
lxcl ,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
ii is ooly rcasonrblc lhsl rls€8rcb.fi i! lhe ficld of biocsttlysb I wel as dccisi{tn
don€srically,
drsters siut to lhiDl about this pmbl€t!" Itr lldoncsi4 wherr i[dultries arc not yct cloccty liokcd wilh
tbe res€€Irh srd scadthic world, a€s€a&h prodlcts would al*rys b€ mercly rcsearch ptodualr ulless
thc d.cisioo rmt.rs tssdt tbciruulbority in policy Ir|AkiDg, supPortiDg thc .pplicstion of &tmcsdc
rH.arcb Fodwrs in odion l hd$ai€s" Tb€rdor€, in or&t lo bring rbe indusEirlists' cdryne
cscrchcrs and dccision natcr closcr togelher, wc orgaoiscd ltc Coofccocc oo bdrstritl Eszfne
Biotcchmlory 2010 (C!EB 2OlO), which wrs hcld iD Targrtalg otr 3 - 6 August 2010' wftt ritlc
EtrzyDe T.chlolo$/ for f,.!-Frlctdly IDdutfy.
T|llcr|rg,3
Angutt 2{10
Dir.ctor of
Cent€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
List ofcoolenl
lti
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
C\lIrlnt
Stste
ofEtrzync R€scarch fu Indon€sia
Id.ltification ofTb€dmphilic C€Uulasc-Ptodrcing Bactdia fiom M€rap
kolation
aDd
Volcm
Craler ln Wcst Sum3t€ra
Jantssri. YusniwEti, Aswaldi Anr,sr, B€Dtri Ismul
PA
tl!
Suhiiooo
Massy Tbenawidjajr
P4l
|!
Willte
Huda
I
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'
P43
7
Scllaritrg lor cellutasc Foducilg b€ctcria lrom Talifugu n bripes fish
Diatr Andtiad, Bambabg Pras€tya, Don H€e
P{4
S
Pa*
l7
Scr! itinS ofDt€rmstrblc Cellulrs€ tltd Xihnase Producing Microo.gtntutB
Prima Endaag Susilovui: S€plo RrharF; ArdiaDsysb, Rthrmwad Rthirq sami
Mrreeti, Ahnd
P45
Selcctiotr ofTr}?cin
thc eozyne
litc
tuclcria atrd cbaErlcriation
Wahlutsri
of
30
Clh$iclcrizarbn of Allrlo-themrybilic XylaDss. Prodrrciry Bsdeda boLt€d from
Rokrn Hulu, RiaD
Shafs Noor,
P4?
rid
proteasc p.oducing lactic
Dysh Wutatrrari, Budisih
P46
A
Zsctri
Ytineri*arni;
Trismilth
ScrE iting of EDzyne Aclivity
t7
amng Fuogal
and
BeLrial
Isolatcs as
Biocoatol
Agent ofcbbroot Dis.asr (PlasDodiophora brossicac) on Ctbbegc (Br&ssicr
olcraccae).
Bahsruddin, Tutit Kuswintnti aod
P-08
Xylmligo6scchtridd
fuu.tion
Cusmiaty
17
ftorn Corncob Xylan uting Sttptotnyc6
co6t rictolls 451-3 Enzy.nc.
Adr Mctyaoditril, Titi Csdra Suttni2,
P-09
EDma Msutta
Moko3
52
Pote irsl ofAlgd BionDss for Biodicsel Prcducing Thtougb Etrzynrtic
Tralr.st rification olMicmalgal Filty Acids
JoLo Sulirtyo, Ritr
D*i
Rrbayu aDd Yati Sudtryiti
Sockr
a
cdnffi d
P-10
rhd6tn.l E.:wR .nd srd.cnn
ld'
lcrai
20101
'Erffi T6htlo'v fd feFdadlv lldr6l-
OprinrlizatioD ofcbiliDsse Foducrio, by B.cilus
5P
Cl5
and chtr"act€dzrlion
3
AId
2010
oftbe
Ruby S.tisctsD, Budirsih Wshyutrttri
P-l
I
Optimizstion ofAcration rnd Cooubstntc addition in Xilosc Bioconvcrsion hto
Xiliiol by Cadi& Ttopicllis
I*smi
P-12
Anborssri, SutyaDi, FraDsislo
Optiniution OfNa-Alginrtc AItd C!cl2 CoFttrtrstion For lromobiliz.d Cc[ Cmdids
Tropicalis Itr Xylitol Ptoduction
Surysd,
P-13
Irtsmi Amb€.srri,
Dyts HisrDysnti
Tbc rfr€ct ofT.ichl,orfon m Cbolircst r.sc Activity of Difietltrt OrgrDs
Musscl, Mytilus ed is.
L
P-14
Elr tludaFni
ofBhE
Ysoio aDd P--D. HtDlcn
lhc Efrcct ofYoghutl Th.mpy on Pmfilc and Activig ofDigestive Ellz''EG (PmE"se,
Lipasc rd suc1..se) Expc.d by Fonnrfdehy& or Rttrus iov€fgiq$ Jejuum
l0l
ftr RlhErtul tryly, Aulami'an, ChstrifMthdi
P-
I
5
Thc lrtrp.ovement of L-srabino6e isomcrase local Gcob.cillus stearolhctttbphilus
Acidic tolerarce for Tagalose hdulbirlizstion by Sit€-Dfucct€d Mutag€ocsh of Glu269
Dcwi Fitriad, Lita Trirsbr atrd Budi
P-16
125
CeD€ ofBs.illua 3p Ar-5r:
and Ctaracletizriion ofthc producl
Cyclodcrt in Gtyco6yl Trrnsfcnse Synthaic
OveExpr€ssion iD Eschcrichia
Catur Rirni,
P-l?
S€Isono
c.li
Alhnsd Muchlisin, Casilawsti, Debbie Sotie Rehodngrum
130
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
P-18
P-21
Widrda
Phstrctochaetechrysodporiurn
(PcCDtI), Substrale Atrtrity,
Reconbinrnt
Debydrogentsc
Flavin Domain Celobiosc
Optinum pH ad Storsgc Stability
SODE
t5,,
Atplicarion ofEnzym LiDtcd lrDtDuno6oftcnt Assay (ELISA) for hrlv Dete.tion
PLRV, P\D( ad PVY on Atlantic Varicty Potalo s€€d G€n€mtion 0 (C0) and
Genedtiotr 2 (G2)
of
Tutit KuswiMnti. A. Mtsdia*sti, Adc Rosnstra dsn BahaDddin
Th€ Us€ of Mol€culsr Tccbdque in Cath€cin Corcentrsiion Diagnocis of Ur.ottt
grndir Roxb
Istioo Fcri6', JaDsari, Irfan Suliatrryrh, Austis4 Hamda Fsuzg dan
I
14
CbanclcrizationofBioclcctochcmicrlElecEod€of
D$riari, Slcftm FERRT Koji
P-20
l35
Cloning OfTbernosbbl€ Dna Polymqsee C€tre Fron A Thqmophilic Br€vibaciuus
Sp. IBolal€d Fmm Sikidary Cmlsr, DcrS Plalar, Ccdtnl Java
Lucir Dhiaditr Witsssri, kfrtr Dwija Prijrobsda! Jafs
P-19
Wibowo
ConEspo.dios.'nhor, itrinofciit'@Fboo.cot'
Yusniwati
155
150
PA2
D.tectiotr of Aedd r.S/pti ard Ac. ilbopictB Vins h Deryuc Fever EDdanic ilt
Prdug City using Rcvcts€ Trsmciptala Polyrretasc Chain Rcaction Mclhod (RT-
rcR)
Hssmiwati, D.bclmi, Nurhayati
P-23
Por.
ial for Protein Dcposition in
161
Diffcrr
Btoile. Ccnotypcs Dependiry on Age
t16
Sarnadi
Pt4
l.ovaslatia Cooteft Produ.lion By Mut tr Offupergilus Tcnclrs Using Solid
FcfirEntation Procrss Atrd Liquid Fcllicnt'liotr Prccess
Triatr' Rias Dewi, Adc SuDiatdi, Susi
P:25
ltl
Effect OfFemcDtation With Addition Slrrbio Probiotica OD Nutsient Conldt OfRice
Straw (Oryza Sativa L)
A& sumiardi, Suli
P:26
Putwasih
Hctinswari
S)orE Mclhods to Detlct lh. Ecotoxic
190
of
Agroch€micrlsotr Mictobidl Activity io SoiI
FcrbbatrTirdaotr
Pt1
198
Tbe Uptrke ofcadmium loos md lls Influance on Thc Cro*'tb and hoduciiotr
Somc S€codary Mcbbolitcs in Shoot CultuEs of Solfllum mcloogcna
of
TjicKok
P-28
2lls
Ctdlicsl Caqo6itioo OfEss€otitl Oil Ofsouth
Surnatran Pittosporufi
FenugiEun
L AiI.
Ad€k ZntDrud AdnsD, Z{iouddin,
ORCAl{IZINC COMMITTEE
Asmtdi
Zls
216
cdl@
r{'d!&!tcEr
d ftnd, !m
toiol'kEt&Y
ntF^atti'db bddl. I Adtolo
P-27
The UPtrke ofCrdmlum lonr rnd 16 Inlh€nce on The Grcwtl rnd
Producdor ofsome Sccondrry Mctrbolltec In Sboot Culturcr ofso'4T |,'t
mdorScng
TJb t(o|('
Fedty
of Biot
cholo$t, Mrrt oatica
aDd Scicacc
Dqcrb'ot
Univ€tlity of surshoyr' susb6ya' IDdmcoia
A'STRACT
*!3 to ctaniDc lhc tltpoBc givco by shoot cuhr€ of &'l@ild't tttlo'Be"a
rac.rch' it could bc sarad thst s&oor odturE
Woo rpt ti|tt ofclddiun iotr& Fm ltc rledls of ttfu
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 tM
cldmioro ioos lo thc E dis cau.d r dacc.rc b lolrl fte phyroEolr c6t nt (50.2y.) of ltc
Tbc sim of this $|rdy
culotr!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|cod
tld t of tbc hrDiDg of f6sil firb sd dltridFl w!rt6. Howcvcr, th.rl
bv. b.co dbl€ ins@6 of loxicity I tbc |lsolt of lonSi'r.tm cxpdn€ lo ctdmiud iD
cadmiud
rI! Firrldly
contaDriDrod food aDd
lhc
*d€r.
ID thc dacrd€ follosing
ll&dd-Wa!!, Jqatrcs.
ttritritr8
op.f,.lid3
!!d Esc6 of olhcf ioxic m.l8ls. Ar a conlcqu.!.q
cootrmiDrrcd thc Jin ! Rivcr with crdmium
cadroium 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@l
lhdo@liti:s, itrcftdiDg plalgulr! tDd gluc@ri& Cldnium is oe of six substaoc€ b.DDcd by thc
EurtDa€! Unioob Rrrtrdion
coqot
m lt zrtdous Subct|rc€s ditEcrivc, wticb h.o3 carciloSlat
m
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?le
ziDc iD may biologicd syst@, io Frricular, rys&43 that codtai! son.f [g.Dds $ch .. adfin
Cad|nium c.tr biod rp l,o tao tim ootl rtugt tbr zioc b c.rtti! biologfu.l tystd, aod i8
mtoriously difFq l to rcmvc. h ldditioo, c{dlsiui! cro rtple maqnesium sDd lglgigE in octlsiD
biotogicd iFt6E3, rlthoogh th€sc rtpltcctrots u! rtrc (AnooiE, 2mt).
Corft .ponding &!hor, [email protected]
208
Phrt ccll culturcs, ss wcll.s irs hrrcr phDI crn give ccrtair rEsponscs on thc lrlsarce of ccrtain
mct&l ions in thc medium. Cltlnrcs of Chlorc a wlgaris could lelnov€ 75% of 8 UM coppfi ion6
fion lhc tn€dium (HemsD, 195). Hairy rool culturB of An &racia n/Jti.ona coluld rennve 62.4./0 of
lhc coppcr ioDs tom initial \Eluc of 330 FM copp€r ioss (Zilrurndova 1995). CopE r tolEa calbs
(-,ulfircs oI Nicoaiorra tab@utt rcge&rated pl"trls thal we.E able to gro$r iD thc prcacnc. of 100 FM
coppef ior|s (cori ct 31., 1998).
Tbc Fodu.aioD of scrquit
rp@id ph)loslcxitri itr ccll silpctrsiotr qrlti.es of Lhttra starnonnort
ad Wtiteh€a4 1989).
bave b€cn itrdrc.d by mctal ions, iDcluding cad&ium ions Clhrcfsll
i
Th€ rEsulls of prcccdhg rcs.archs denoost-ald fiat
roducing rEtal iobs, iDcMitrg crfuium roos,
into pletrt cell culNres could inducc, incrcas€, or iohibit thc goductiotr of sorra sccondary mctabolitcs
in that cullur€s.
E'OERIMENTAL
l,tateriab
Thc shoot cuhur.s oflo/@irr tttclongern wer. cttltivsl|.,l od rmdificd Mu.sshige ltrd StooS nedit
with lhe addition of 2 mg.L ' bcnrylsdenin€. Cultures were rnaintainrd h the ligbl using philips
TL40w 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€not
corrc.ntratiotr of cadmium iotrs sDd cultivatd for 4 w€€ks udcr tb€ saoc conditiod prior to
harvcstilg. For each csdmiuh iotrs conccntsatior! 15 botd€s of +l-l.j gram fr€sb weight of shootr
cach 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
w.i8nt
rd
Analysb
h grodrr t der (CD, dcfm.d as the ratio of final fiEsb
iaitid filsh weiSbl of.ach culrre. Th. CI r,!s msurod for cach bottle.
ofcodttin
iont
Cadiium ions in the biom.ss wc& dctermined by using tCp-AES aftd apFoprial,, digestion of 0.1
mg powd6 using 2.0 ml coocedtrsted HNq fouow€d by dihxion with dirtijld wrtei uD io 10.0 ,nl or
dilutld fimher as r€quircd {Cb€n, t993}. qrbd6catjon ws! pcrfonDcd by me{surio; dle crtrlssim
itrtcosiry ofcrdmirtu rr ib maximum *lvclcngth. The coocentration ofcsdmium iotrs \lr.s calculstcd
fton the lio€a. rcgrrssim cquaiion ofsiandardRf,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
sucrGe cooccrtsetioD in mcdia bcfore add affer hsrvesting w€r€ sbown oa Tsblc t , FigutE I aDd 2.
of
2(B
co.{Ru d
Tabfe
L
hdu*nd
Eh:ft ..d sdxn.oldv
lcrFs 20101
'|ff.
Tdhmlle fc E@FndnN bd's'. I Aud l0l0
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
D€slasc of
ConcetrFation
ofcd"
Replication
sucaose
CroMh
ind€x
(Fl'f)
('/"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
4_58+1.84
24.51+0.55
t60
The gto*'th i[dcx wt3 cxpc*scd
o/o
avemce + devirlioo statdard.
s
av6rga + d€vialion staodar( cbnngp in $E osc cooccntsation as
9.(D
6.tb
7.00
6m
5.CD
4.00
3.00
2.m
1_q)
0.00
80
coldffi..ac'n+
1m
(!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
8o
120
Cmtd-dcd'+(rM)
Figure 2. Graph of lbe decrcrx€ of sucaos€ corccntrstio!
with variatioo
conccoEarion
ofc?
I
i! medit
a
Figur€ I $owed thalth€ sioot cultues of Solan/n ,Elongenz \|ere sble to grow in the
mcdfu contrining cadmium iotrs coaccntmtion up to 160 IM with lbe gD*tb index grratcr thao thr€e.
Thc growlh lal! ioq€aled witb lbc ioc'rrosiry cnnc.otratioD of cadmium ioos, r@h€d ils maximur! sr
Table
ibe conc€aEation of rpproximdely 120 pM atd dccrcascd anErw!.ds. It was sactr ftom Table
I,
Figures I ad 2, rhat tbe growth rate was not olways invefcly proportiooal to rhe dcc-rease of sucnoce
concentralion ir ocdi& lt wes possibly due to tbe diff€Encc itr stEorption of nu[ie|nt otb.r tbm
3UCr6€ by lbc Cultrcs.
Analys is oI p ht
tos,e ro I (s)
Tbc rrsults of tbc a$\sis of stcroidsi Slyco6idca(c) usilrg .nisald€iydc.Hzs0. .s s sprsyidg rlsgedt
w€rt sbowD on table 2,
Trble 2. Alrlysis ofs{croidsl glyco6idds) using titr hycr ct ornatography Cn C}
Concetr!!lion
ofcd'(rM)
zt0
80
120
160
St€roidal glycoside(s)
Resull
Rf
0.63 (ete.n),
0.71
0.63 (rrEen),
0.71
(ycllow),0.63 (violel)
(ye[ow],0.81 (violet)
0.63 (g€€tr),
0.71
0.63 (seen),
0.71
(yellow). 0.83 (violei)
(yellow). 0.81 (viol€r)
+ indical€s that s substance being id€ntified is existing, Rf is ret6rdation factor
211.
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/arut
tnclongata 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@r
rrelangeru cultur€s aged 4 w€eks
Concctruali,ot
Corceo!-ation of Conccntrtiotr of
cd' h bionrss totd fie€ sterols
(ltziP\
lqc,/c.:)
ofcd"
Replication
40
t5
3.6+02
80
52.0+1.4
568,!98
450+75
t20
l5
l5
160
t5
101.2+4.6
135.9+2.1
324+87
(nM)
Concent_atioD ofcadrnium ioDs aod lotal frE€ stcmls
*ls
421+g)
expressed
in,4/g dry weight ofbiornsss
150.0
125.0
1oo 0
75.0
50.0
25.0
00
a{)
80
c...-d
120
.a
cdt+ (rM) h
t6t)
..d.
Figurc 3. cr+h ofcoratnrion ofcd:+ in bionass
with variation ofC
00il0
Gotrf.r.nco on !ndustd.l Enzth. .nd Elot.chnologtf
Proceeding
Enzyme Technology
Serpong, Tangorang
Seminar, 3 August 2010
j
t
'1F
tra
.
Flz!'
ilaLr
E!
'l',
,-
I
Held by
$)
RTSTEK
PUSPIPTEX
Deputy ofTechnolow toa Agroindustry and Bioteclnol
Agencyfor the Assessm€nt d|d Application of Tecrnobgy
Suppoftd by
cdt r.E n hdsti.l E|lm
rnd
g,ti.dnoldvlots r6{n'F;'w Trlmloft fe EsFndlv hdr6a.1a|d
2OlO
ISBN 97&979-9U04-9-3
PROCEEDING:
CONFERENCE ON INDUSTRIAL ENZYMES AND
BToTECHNOLOGY (CIEB 2010)
Enryme Technology
for Eco-Friendly Industry
Orgrrlr.d byl
CeEter for T€chnolos/ of Blo hduttry
Goverrrn€nt Agency for Acsgamert .trd Appllcrtlor of TechDology
itr cooParadon with
R6e.rcb Ccller for saleDc{3 rDd Techlolos/
MlDlrtry of Relc|rch rrd Tecbtrolog/
rnd
IteutlcherAtrdemilchcr Au*rNchdlcnat (GerDrtr AcadeEicErcbrtrge S€rvice)
T.nSer|
g' 2010
cdtr@d
l"d,nti.l Elrm.,n €ldldtn
ro.v
lotl to10l'Fid T.dftldv tr
FaFnadrY
I ||d'.I
tu 2rlo
ISBN 978-979-9U04-9-3
PROCEEDING:
COI\IFIRENCE ON INDUSTRIAL ENZYMES AI\ID
BIOTECENOLOGY (CIEB 2010)
Enzyme Technology
for Eco-Friendly Industry
Tangerang, Brnten 03 August 2010
Edibd by:
Budiasih Wahlrntad
Suyanio Pawiroharsono
Sise.a Setyrhadi
ls Hclia$i
Trismilah
@
All nght r€served
Publish€d bY:
Center for Technolog/ of Blolndwtry
Government 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
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 flrlticellular
orgadisrds. Enzyn€s phy 6n ir4ona role in our live, not only in our oEn hctlbolism, but also in
o|h€i proccss€s susrlhing our dsily activiti€s. Thcy are us€d lo sssist tll€ productiotr of th€ food *c
cat aDd contribule to the carE of ou. healh by lroviding tb€rspeutic aSenls or scBitive a.d sp.ci6c
dirgdo€tic t .{s. Thus ir tss hid conerci.l rd indusr.ful Yahrc.
Enzym€s
Enzytrs also play atr irDponrot tolc in 'gftcn indstry'. lDrtrtd of using mor€ hszardous
c rhore envitonrnentslly fti€ndly choic€. For cxerqle, ifftead of
using cblorinc for tbc bldhing p.ocess€s, ptot ax. is a 'gt€toet' choice for detcrgrd as wells pulp
cbcmicals, €trzyrmtic proccss is
asd
paF
hdlrstri€s.
oow in higi6 dcmt s, dr€ to
In additio& prodll;ls oore cnviotrmtrttlly fticrdly proc6s.d
'It C6s€s involviry rcjeclio[s of
tigbt r €nvironrFntrt rrgulstions, elpccialy in thc dcvclop€d colmni€s.
hdon.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.d
si.ic-t r l0r,i rrrd h5d tbc iDdustislisb hsd rmtE lwarEn€ss oo eDvironDnld issrB EDrymd'
tbercfcr€, would b€ otre ofth€ answ€ts.
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 with
industial coryrns sd 253 US do[rrs wilh spccirlty enzyms us€d in tle pbsmac€utic,L te*"tch
aDd di.gtro*ic field6. Atdougb lbc Ftrctr k lcrs pEdictablc, IDdo.si.tr dorncstic d.mrn& s.dr ro
The drta
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 .s
lxcl ,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$i& as wcll 3s
ii is ooly rcasonrblc lhsl rls€8rcb.fi i! lhe ficld of biocsttlysb I wel as dccisi{tn
don€srically,
drsters siut to lhiDl about this pmbl€t!" Itr lldoncsi4 wherr i[dultries arc not yct cloccty liokcd wilh
tbe res€€Irh srd scadthic world, a€s€a&h prodlcts would al*rys b€ mercly rcsearch ptodualr ulless
thc d.cisioo rmt.rs tssdt tbciruulbority in policy Ir|AkiDg, supPortiDg thc .pplicstion of &tmcsdc
rH.arcb Fodwrs in odion l hd$ai€s" Tb€rdor€, in or&t lo bring rbe indusEirlists' cdryne
cscrchcrs and dccision natcr closcr togelher, wc orgaoiscd ltc Coofccocc oo bdrstritl Eszfne
Biotcchmlory 2010 (C!EB 2OlO), which wrs hcld iD Targrtalg otr 3 - 6 August 2010' wftt ritlc
EtrzyDe T.chlolo$/ for f,.!-Frlctdly IDdutfy.
T|llcr|rg,3
Angutt 2{10
Dir.ctor of
Cent€r for BioiMustrial Techtrolog/
Wilooo B&suli
Conf.r.M. m rndunrEl
t.sm. ..d
Bbr..Jlnolon lcrFs
2or0l'Em Tajlrollp fn E*Fnddlt lnd!6/ . I Auo< 2010
Co[teDa
P18€
Introductios
List ofcoolenl
lti
iv
IndBtrinl Etrztme Prcduction ia B&t.ti.: C,.netics rnd Sttlitr lopmvdEnt
Friedhcta
Meinhardt
vlll
BioconvcrsioD: From BioE ss lo Cbcnicsls atd Plod.rts
Klnus-Dicter Vorlop, Frsn! Scbucbrdt, Hcinz SticbDothc, Tboms
C\lIrlnt
Stste
ofEtrzync R€scarch fu Indon€sia
Id.ltification ofTb€dmphilic C€Uulasc-Ptodrcing Bactdia fiom M€rap
kolation
aDd
Volcm
Craler ln Wcst Sum3t€ra
Jantssri. YusniwEti, Aswaldi Anr,sr, B€Dtri Ismul
PA
tl!
Suhiiooo
Massy Tbenawidjajr
P4l
|!
Willte
Huda
I
hohtion OfThernostsblc -Amyhse From Th€mophilic Ba.l€.is Of Soiai Hot
Spring SouIb East Sulawcsi
Sspto R.ahrrjo, Andi Noor Kholidhssysrifin, L.M. Rtntdben, Prima Endang'
P43
7
Scllaritrg lor cellutasc Foducilg b€ctcria lrom Talifugu n bripes fish
Diatr Andtiad, Bambabg Pras€tya, Don H€e
P{4
S
Pa*
l7
Scr! itinS ofDt€rmstrblc Cellulrs€ tltd Xihnase Producing Microo.gtntutB
Prima Endaag Susilovui: S€plo RrharF; ArdiaDsysb, Rthrmwad Rthirq sami
Mrreeti, Ahnd
P45
Selcctiotr ofTr}?cin
thc eozyne
litc
tuclcria atrd cbaErlcriation
Wahlutsri
of
30
Clh$iclcrizarbn of Allrlo-themrybilic XylaDss. Prodrrciry Bsdeda boLt€d from
Rokrn Hulu, RiaD
Shafs Noor,
P4?
rid
proteasc p.oducing lactic
Dysh Wutatrrari, Budisih
P46
A
Zsctri
Ytineri*arni;
Trismilth
ScrE iting of EDzyne Aclivity
t7
amng Fuogal
and
BeLrial
Isolatcs as
Biocoatol
Agent ofcbbroot Dis.asr (PlasDodiophora brossicac) on Ctbbegc (Br&ssicr
olcraccae).
Bahsruddin, Tutit Kuswintnti aod
P-08
Xylmligo6scchtridd
fuu.tion
Cusmiaty
17
ftorn Corncob Xylan uting Sttptotnyc6
co6t rictolls 451-3 Enzy.nc.
Adr Mctyaoditril, Titi Csdra Suttni2,
P-09
EDma Msutta
Moko3
52
Pote irsl ofAlgd BionDss for Biodicsel Prcducing Thtougb Etrzynrtic
Tralr.st rification olMicmalgal Filty Acids
JoLo Sulirtyo, Ritr
D*i
Rrbayu aDd Yati Sudtryiti
Sockr
a
cdnffi d
P-10
rhd6tn.l E.:wR .nd srd.cnn
ld'
lcrai
20101
'Erffi T6htlo'v fd feFdadlv lldr6l-
OprinrlizatioD ofcbiliDsse Foducrio, by B.cilus
5P
Cl5
and chtr"act€dzrlion
3
AId
2010
oftbe
Ruby S.tisctsD, Budirsih Wshyutrttri
P-l
I
Optimizstion ofAcration rnd Cooubstntc addition in Xilosc Bioconvcrsion hto
Xiliiol by Cadi& Ttopicllis
I*smi
P-12
Anborssri, SutyaDi, FraDsislo
Optiniution OfNa-Alginrtc AItd C!cl2 CoFttrtrstion For lromobiliz.d Cc[ Cmdids
Tropicalis Itr Xylitol Ptoduction
Surysd,
P-13
Irtsmi Amb€.srri,
Dyts HisrDysnti
Tbc rfr€ct ofT.ichl,orfon m Cbolircst r.sc Activity of Difietltrt OrgrDs
Musscl, Mytilus ed is.
L
P-14
Elr tludaFni
ofBhE
Ysoio aDd P--D. HtDlcn
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
l0l
ftr RlhErtul tryly, Aulami'an, ChstrifMthdi
P-
I
5
Thc lrtrp.ovement of L-srabino6e isomcrase local Gcob.cillus stearolhctttbphilus
Acidic tolerarce for Tagalose hdulbirlizstion by Sit€-Dfucct€d Mutag€ocsh of Glu269
Dcwi Fitriad, Lita Trirsbr atrd Budi
P-16
125
CeD€ ofBs.illua 3p Ar-5r:
and Ctaracletizriion ofthc producl
Cyclodcrt in Gtyco6yl Trrnsfcnse Synthaic
OveExpr€ssion iD Eschcrichia
Catur Rirni,
P-l?
S€Isono
c.li
Alhnsd Muchlisin, Casilawsti, Debbie Sotie Rehodngrum
130
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
P-18
P-21
Widrda
Phstrctochaetechrysodporiurn
(PcCDtI), Substrale Atrtrity,
Reconbinrnt
Debydrogentsc
Flavin Domain Celobiosc
Optinum pH ad Storsgc Stability
SODE
t5,,
Atplicarion ofEnzym LiDtcd lrDtDuno6oftcnt Assay (ELISA) for hrlv Dete.tion
PLRV, P\D( ad PVY on Atlantic Varicty Potalo s€€d G€n€mtion 0 (C0) and
Genedtiotr 2 (G2)
of
Tutit KuswiMnti. A. Mtsdia*sti, Adc Rosnstra dsn BahaDddin
Th€ Us€ of Mol€culsr Tccbdque in Cath€cin Corcentrsiion Diagnocis of Ur.ottt
grndir Roxb
Istioo Fcri6', JaDsari, Irfan Suliatrryrh, Austis4 Hamda Fsuzg dan
I
14
CbanclcrizationofBioclcctochcmicrlElecEod€of
D$riari, Slcftm FERRT Koji
P-20
l35
Cloning OfTbernosbbl€ Dna Polymqsee C€tre Fron A Thqmophilic Br€vibaciuus
Sp. IBolal€d Fmm Sikidary Cmlsr, DcrS Plalar, Ccdtnl Java
Lucir Dhiaditr Witsssri, kfrtr Dwija Prijrobsda! Jafs
P-19
Wibowo
ConEspo.dios.'nhor, itrinofciit'@Fboo.cot'
Yusniwati
155
150
PA2
D.tectiotr of Aedd r.S/pti ard Ac. ilbopictB Vins h Deryuc Fever EDdanic ilt
Prdug City using Rcvcts€ Trsmciptala Polyrretasc Chain Rcaction Mclhod (RT-
rcR)
Hssmiwati, D.bclmi, Nurhayati
P-23
Por.
ial for Protein Dcposition in
161
Diffcrr
Btoile. Ccnotypcs Dependiry on Age
t16
Sarnadi
Pt4
l.ovaslatia Cooteft Produ.lion By Mut tr Offupergilus Tcnclrs Using Solid
FcfirEntation Procrss Atrd Liquid Fcllicnt'liotr Prccess
Triatr' Rias Dewi, Adc SuDiatdi, Susi
P:25
ltl
Effect OfFemcDtation With Addition Slrrbio Probiotica OD Nutsient Conldt OfRice
Straw (Oryza Sativa L)
A& sumiardi, Suli
P:26
Putwasih
Hctinswari
S)orE Mclhods to Detlct lh. Ecotoxic
190
of
Agroch€micrlsotr Mictobidl Activity io SoiI
FcrbbatrTirdaotr
Pt1
198
Tbe Uptrke ofcadmium loos md lls Influance on Thc Cro*'tb and hoduciiotr
Somc S€codary Mcbbolitcs in Shoot CultuEs of Solfllum mcloogcna
of
TjicKok
P-28
2lls
Ctdlicsl Caqo6itioo OfEss€otitl Oil Ofsouth
Surnatran Pittosporufi
FenugiEun
L AiI.
Ad€k ZntDrud AdnsD, Z{iouddin,
ORCAl{IZINC COMMITTEE
Asmtdi
Zls
216
cdl@
r{'d!&!tcEr
d ftnd, !m
toiol'kEt&Y
ntF^atti'db bddl. I Adtolo
P-27
The UPtrke ofCrdmlum lonr rnd 16 Inlh€nce on The Grcwtl rnd
Producdor ofsome Sccondrry Mctrbolltec In Sboot Culturcr ofso'4T |,'t
mdorScng
TJb t(o|('
Fedty
of Biot
cholo$t, Mrrt oatica
aDd Scicacc
Dqcrb'ot
Univ€tlity of surshoyr' susb6ya' IDdmcoia
A'STRACT
*!3 to ctaniDc lhc tltpoBc givco by shoot cuhr€ of &'l@ild't tttlo'Be"a
rac.rch' it could bc sarad thst s&oor odturE
Woo rpt ti|tt ofclddiun iotr& Fm ltc rledls of ttfu
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 tM
cldmioro ioos lo thc E dis cau.d r dacc.rc b lolrl fte phyroEolr c6t nt (50.2y.) of ltc
Tbc sim of this $|rdy
culotr!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|cod
tld t of tbc hrDiDg of f6sil firb sd dltridFl w!rt6. Howcvcr, th.rl
bv. b.co dbl€ ins@6 of loxicity I tbc |lsolt of lonSi'r.tm cxpdn€ lo ctdmiud iD
cadmiud
rI! Firrldly
contaDriDrod food aDd
lhc
*d€r.
ID thc dacrd€ follosing
ll&dd-Wa!!, Jqatrcs.
ttritritr8
op.f,.lid3
!!d Esc6 of olhcf ioxic m.l8ls. Ar a conlcqu.!.q
cootrmiDrrcd thc Jin ! Rivcr with crdmium
cadroium 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@l
lhdo@liti:s, itrcftdiDg plalgulr! tDd gluc@ri& Cldnium is oe of six substaoc€ b.DDcd by thc
EurtDa€! Unioob Rrrtrdion
coqot
m lt zrtdous Subct|rc€s ditEcrivc, wticb h.o3 carciloSlat
m
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?le
ziDc iD may biologicd syst@, io Frricular, rys&43 that codtai! son.f [g.Dds $ch .. adfin
Cad|nium c.tr biod rp l,o tao tim ootl rtugt tbr zioc b c.rtti! biologfu.l tystd, aod i8
mtoriously difFq l to rcmvc. h ldditioo, c{dlsiui! cro rtple maqnesium sDd lglgigE in octlsiD
biotogicd iFt6E3, rlthoogh th€sc rtpltcctrots u! rtrc (AnooiE, 2mt).
Corft .ponding &!hor, [email protected]
208
Phrt ccll culturcs, ss wcll.s irs hrrcr phDI crn give ccrtair rEsponscs on thc lrlsarce of ccrtain
mct&l ions in thc medium. Cltlnrcs of Chlorc a wlgaris could lelnov€ 75% of 8 UM coppfi ion6
fion lhc tn€dium (HemsD, 195). Hairy rool culturB of An &racia n/Jti.ona coluld rennve 62.4./0 of
lhc coppcr ioDs tom initial \Eluc of 330 FM copp€r ioss (Zilrurndova 1995). CopE r tolEa calbs
(-,ulfircs oI Nicoaiorra tab@utt rcge&rated pl"trls thal we.E able to gro$r iD thc prcacnc. of 100 FM
coppef ior|s (cori ct 31., 1998).
Tbc Fodu.aioD of scrquit
rp@id ph)loslcxitri itr ccll silpctrsiotr qrlti.es of Lhttra starnonnort
ad Wtiteh€a4 1989).
bave b€cn itrdrc.d by mctal ions, iDcluding cad&ium ions Clhrcfsll
i
Th€ rEsulls of prcccdhg rcs.archs denoost-ald fiat
roducing rEtal iobs, iDcMitrg crfuium roos,
into pletrt cell culNres could inducc, incrcas€, or iohibit thc goductiotr of sorra sccondary mctabolitcs
in that cullur€s.
E'OERIMENTAL
l,tateriab
Thc shoot cuhur.s oflo/@irr tttclongern wer. cttltivsl|.,l od rmdificd Mu.sshige ltrd StooS nedit
with lhe addition of 2 mg.L ' bcnrylsdenin€. Cultures were rnaintainrd h the ligbl using philips
TL40w 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€not
corrc.ntratiotr of cadmium iotrs sDd cultivatd for 4 w€€ks udcr tb€ saoc conditiod prior to
harvcstilg. For each csdmiuh iotrs conccntsatior! 15 botd€s of +l-l.j gram fr€sb weight of shootr
cach 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
w.i8nt
rd
Analysb
h grodrr t der (CD, dcfm.d as the ratio of final fiEsb
iaitid filsh weiSbl of.ach culrre. Th. CI r,!s msurod for cach bottle.
ofcodttin
iont
Cadiium ions in the biom.ss wc& dctermined by using tCp-AES aftd apFoprial,, digestion of 0.1
mg powd6 using 2.0 ml coocedtrsted HNq fouow€d by dihxion with dirtijld wrtei uD io 10.0 ,nl or
dilutld fimher as r€quircd {Cb€n, t993}. qrbd6catjon ws! pcrfonDcd by me{surio; dle crtrlssim
itrtcosiry ofcrdmirtu rr ib maximum *lvclcngth. The coocentration ofcsdmium iotrs \lr.s calculstcd
fton the lio€a. rcgrrssim cquaiion ofsiandardRf,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
sucrGe cooccrtsetioD in mcdia bcfore add affer hsrvesting w€r€ sbown oa Tsblc t , FigutE I aDd 2.
of
2(B
co.{Ru d
Tabfe
L
hdu*nd
Eh:ft ..d sdxn.oldv
lcrFs 20101
'|ff.
Tdhmlle fc E@FndnN bd's'. I Aud l0l0
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
D€slasc of
ConcetrFation
ofcd"
Replication
sucaose
CroMh
ind€x
(Fl'f)
('/"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
4_58+1.84
24.51+0.55
t60
The gto*'th i[dcx wt3 cxpc*scd
o/o
avemce + devirlioo statdard.
s
av6rga + d€vialion staodar( cbnngp in $E osc cooccntsation as
9.(D
6.tb
7.00
6m
5.CD
4.00
3.00
2.m
1_q)
0.00
80
coldffi..ac'n+
1m
(!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
8o
120
Cmtd-dcd'+(rM)
Figure 2. Graph of lbe decrcrx€ of sucaos€ corccntrstio!
with variatioo
conccoEarion
ofc?
I
i! medit
a
Figur€ I $owed thalth€ sioot cultues of Solan/n ,Elongenz \|ere sble to grow in the
mcdfu contrining cadmium iotrs coaccntmtion up to 160 IM with lbe gD*tb index grratcr thao thr€e.
Thc growlh lal! ioq€aled witb lbc ioc'rrosiry cnnc.otratioD of cadmium ioos, r@h€d ils maximur! sr
Table
ibe conc€aEation of rpproximdely 120 pM atd dccrcascd anErw!.ds. It was sactr ftom Table
I,
Figures I ad 2, rhat tbe growth rate was not olways invefcly proportiooal to rhe dcc-rease of sucnoce
concentralion ir ocdi& lt wes possibly due to tbe diff€Encc itr stEorption of nu[ie|nt otb.r tbm
3UCr6€ by lbc Cultrcs.
Analys is oI p ht
tos,e ro I (s)
Tbc rrsults of tbc a$\sis of stcroidsi Slyco6idca(c) usilrg .nisald€iydc.Hzs0. .s s sprsyidg rlsgedt
w€rt sbowD on table 2,
Trble 2. Alrlysis ofs{croidsl glyco6idds) using titr hycr ct ornatography Cn C}
Concetr!!lion
ofcd'(rM)
zt0
80
120
160
St€roidal glycoside(s)
Resull
Rf
0.63 (ete.n),
0.71
0.63 (rrEen),
0.71
(ycllow),0.63 (violel)
(ye[ow],0.81 (violet)
0.63 (g€€tr),
0.71
0.63 (seen),
0.71
(yellow). 0.83 (violei)
(yellow). 0.81 (viol€r)
+ indical€s that s substance being id€ntified is existing, Rf is ret6rdation factor
211.
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/arut
tnclongata 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@r
rrelangeru cultur€s aged 4 w€eks
Concctruali,ot
Corceo!-ation of Conccntrtiotr of
cd' h bionrss totd fie€ sterols
(ltziP\
lqc,/c.:)
ofcd"
Replication
40
t5
3.6+02
80
52.0+1.4
568,!98
450+75
t20
l5
l5
160
t5
101.2+4.6
135.9+2.1
324+87
(nM)
Concent_atioD ofcadrnium ioDs aod lotal frE€ stcmls
*ls
421+g)
expressed
in,4/g dry weight ofbiornsss
150.0
125.0
1oo 0
75.0
50.0
25.0
00
a{)
80
c...-d
120
.a
cdt+ (rM) h
t6t)
..d.
Figurc 3. cr+h ofcoratnrion ofcd:+ in bionass
with variation ofC