Immobilization of urease on grafted starch by radiation method

Rodrar

Ph>,

Elsev~er

0969-806X(95)00316-9

IMMOBILIZATION

Vol

C‘hem.

46. No 4-f,, pp. 1037-1042, Science Ltd. Printed in Great

OF UREASE ON GRAFTED BY RADIATION METHOD Nguyen

anh

Dung

,Nguyen

Nguyen

duy

Hang

,Tran

onto

starth

1995 Britain

STARCH

dinh tich

Huyen

Canh

*

**

Abstract rhe biomaterial.The

urease

quantitative

was

relationshlps

determined.The

acrylamide

was

immobilized

between

enzyme

grattrd

ac-tivlty

on

the was

by radiatlon

the

percent

grattrd

gratt

maintained

stnrc h.Somr

and

th(l

sclvrn

is a kind

experiments

nc-tivily

by morc2 thdn

which

to

of immobilized batch

polymeric observe

the

enzyme

enzyme

were

reactions.

Introduction I hr components

mainly

blochcmlcal

fields

derivatles

are

re( ent

by

mean

hav(A

a kind

on

cd

thy

enzyme were

graitcld and

sludle~

and

gratt its appllc

II/Materials Monomer purchased

from

Biological

Institute

levels:5%,10%

group

onto

Cu*+

w(are

(It

a5 their

widclly

(tir)ttm,~n.

anti

gr,+ttt,cl

gootl

c h(arnlc \tarc

bc>lwc-c>n

ol urt’a

th(a

w,i\

079)Thc

alLphysIcal

itarc

iti<

properties

wa\

th(ln

AC t~vity

h and

f)(~rtorrnt~d

and

grafted

method-f

cJn/ymc‘ \I)(‘(

biofunctional

to biomedical

h by radiation

III gr,itt(acl

in \,xrurn

various

applications

h 1 hl, urc.a\(’

h(d I,rol)czrtica<

to analysis

al

t~(

of

I ‘)Hl ,K~c~tsu.l

onto

6’ ot \tArt

ralationshif,\

Immobilization l)ra(

f)o\\cA>s

was

InvestlgatedJ

ations

\tud~~s

as well

the surfac

qualitative

starch as well.

his method immobilized

01 Immobilized

urcAasc,-grafted

starch

as well.

Methods:

acrylamlde

Roquette

was

supplied

(France).Urease

by Schmell)tJrcllic was

enzyme

h M~~rk((;crmany).Maize from

starch

Hiotechnological

was

Center,Tropical

(Vietnam).

Maize (containing

acrylamide

and

the

biomaterial

work

star< h 5orncl percent

studied

oi polymrnc

c arboxylamlclt>

on

of radiation been

In this tntroduc

progress

starch at thcl

and

15%

was

slightly

swollen

IO 2 molar

level).Thc

iw/wJ.-lhe

mixture

111 the

aqurou~

concentration was 1037

put

untic,r

solutions

01 monomer rl nltrogcjn

of was gas

monomer usrd

,irradiated

acrylamide

at three

various

with

various

1038

Nguyen

doses dnd

at dose filtered

rate with

of 0.30

Mrad/hr

distilled

-Grafting

and

water yield

at SS°C

from

M.Alresdasilva water

(Aires

at room

da

temperature

minutes.

The

inilial

time.The

grafted

-Swelling

was dried capatity

Ihr

grafted

and

washed

WdS

weighed

was by

by

(300mg)

were

and

weighed

filtered

graphical

plotting

method

described

immersed every

dnd

in

by

distilled

2 min

over

extrapolation

for

30 zero

wc>lght

rvnludtcJd

by tormula: \Y,

Swellmg tapcIty

starch

weight

determined

starch

star<-h

wa1s obtaIned

to constant wds

grafted

to constant

was

tdpdCity

grattccl

capacity

irradiation,the

by tormuia:

1990).

for Zhrs.The

swelling

starch

et all

After

wo

: swelling

silva

er al

4 to 5 times,dried

was determined

capacity

Dung

0 40 Mrad/hr

WI

-Swelling

anh

WI

PO1 =

x 100

tvi WI lnltlal WC, wt~lglli Wf final dry wrlglll -Immobilization The

immobilization

1 gram and

suspensed

with

5tnrrh

phosphate

(Lowry

buirrr

c-ontrnt et

dl

oi

taken

Flowder buftrlr

tar a ICM. mrnutes

The procedure

grafted

was

In 10 ml phosphaic>

.5600U/g),shaklng washed

I)rocedur’, ~~la( (1 ~5 tallow: LV~< wnsh(ld ~~1lul1017

I h(s
with

100

contdining

Eva\

15 ml phosphate

buffer

mg urease

pul for 4 hours

al room

solution

enzyme

WH=7

(ureasr

lemperature,

filtered

6)

dctivity

and

solution protein

wa\

enzyme

al)\c,rt)vci

onto

grnftrd

stnrch

determined

using

195 1 I

The

activity

of urease

The

activity

of immobilized

wa\ ureasfl

nssayscd enzyme

with was

using evaluated

urea

as substrate by formula

:

Lowry

dSSdy

Yrh International Results

and

1 .tffects The method.The 1.5

samples

Mrads,at

aqueous

dose

solution

was

I able

Meeting

on Radiation

Processing

1039

Discussion:

of irradiated graitlng

dose

and

of acrylamidc

WAS Irradiated

with

rates

Mrad/hr.

ot O-SO

5%. 10%

and

onto various

of irradiated

rakO.40

Mrad/hr.

surface

doses.O.LS

yield

Mrad/hr

obtained

on grafting

of acrylamide

of starch Mrad,O.SO

0.40

The

dose

rate on grafting the

and

15%(w/w).

1: hffec-ts

Dosc(Mrad)

dose

yield

prepared

Mrad,0.75

.The

results

was

starch:

by

irradiation

Mrad.1

concentration

.O Mrad

and

of monomer

are presented

of acrylamide

onto

in table onto

in

I.

starch

at dose

:

1.5

:

:

6.8

(%).

0.25

:

0.50

0.75

51

:

6.4

7.0

1.0

( one cn.t%monomer):

5

10

7. j

:

10.2

0. 5

15

105

:

12.1

IL7

:

the doscl of 0 50 ,Mrad

IO 0.75

I he rc+ult\ graiting 5% 1 Ihe

a(-ryl~midr

monomer gratting

onto

t.ftect that


itarc

c c,nc c~ntr~tlon yield

the percentage

6.5

Inc r(Jr1\(+

that

h At these

doses

.thcd \arnl)lr\

j,
j at 10%

a\ the irradiated

(11 monomer

concentration

of grdtt

obviously

yield

dose

Inc rcases

on grnit increases

tontaln

with

yield

10.0

:

9.7

12.5

:

12.2

Mrad n graft

monomer),1

L..i-1

tram

O.,!5

Mrad

wa5

observed.The

in(.rease

of monomer

are found level

: : :

to be optimal

of 6.4

2 7%

for

% to 7.0%

( al 15%

(at

monomer

to 0 75 Mrad. obtained

results

concentration.(fig.l).

show

1040

Nguyen

anh

Dung

et al.

5

w.!5

O-50

Fig.1

Kelationshll) (XI 5%

Mrad/hr

Mrad

and

tffec-t

oi dose

0..30

Mrad!hr,at

O.l5

,1,01,0,1,,',

rate on gratt thr

l)cJlwtJcAn

same’

~IPIII

4.0 Irradlatrd

UN I LJ '0 ,,,ol,o,~,~r.

dose (q 15%

I hcs t~xpc~rimrnts

mc,ncjrncar

+ Dose ( Hrak)

15

cone
and

grafting

yield.

monomer

were 10%.

assayed Thr

at two obtained

dose results

rates

of 0.40

show

that

9th Inlernational Swelling Iahlr

c aparity:

the swelling

2. I h
Ooscl(Mrntl)

Start-h

dose

at

15

787

106.6

185

01 us(:

and

about

lemprralurf-i

urease

umt\

enzyme

may irom

immobilization

gr

tth(A

he

support

starch

starch

was

at irradiated

07i

presented dose

rate

in table

2.

0.4 Mrad/hr.(%)

1 .oo

:

1.5

:

IS morr

01 ur(Aa\(A

244

0 :

204.6

:

182.3

:

:

220.3

: 188.6

:

:

233.0

\tnrt-h 15%

c-ho
monomer

(I);1 ,SMrads

IO immobillzr

utease

c A~IAC sty ot tht> zarnllles h,iLIng

ot the sample 01 \aml~l(~

content

rl nhsorbent

doscl than

II ,~n,i ~molt~

with

15%

monomer

enzyme. is different.The

(opacity

urfarlsf’

ot over II ha\ oi

1.5

ill 15 low

I c an t)(. rc~l,c~nt(~cily

ot mlcrotnc)l,~r

I and

cJnzyrn,A

:

of 18.4

which % prolein

was and

at

,at tfoic~ 01 1 i bA;lrAti< 15 f, i O”!~,, protein

a( t~v~ty

~II high

5

with

5?,, monomer

01 ~rnmolt~lrzf~~~

\nmplr

Mrad

th(J al,\ortx~nt

with

ur(‘a<(’

gratlc,cl

(III) wns

that

do tlvitb

n rt+l(lurll

Although dliferrnt.11

monomer

Mrntf

onto

of 0.5

I\ OI) .! ‘!i, I’rolcln

A( tlvily

maintain>

at dose

chow,

Immot)ll~/c~ci 150

of urrnse

5’S

I hch rt+itiu~l lime

0 50

171.i

ot 0._‘5

ot 0 5 Mtatl

cl.‘5

9.5-S

with

dosr

graitwi

78.7

I hiA rc>sult\ grafted

OI the grafted

I0

grattecf

0.25Mrari

oi thr

1041

Processing

iYO monomer)

?.lmmol~il~rat~on

(II) and

on Radiation

capacity

c-apa’ity

0.00

(Ions-en

Meeting

II) fi

Nti

u~(i

i wrl\

/II ~IA~~v’\ ~IowIy

and

decreases

quickly

after

has

a residual

activity

of 70 %

minute

al

hydrolyzed And

after

for 7 times

one

one

room

of use ,immobilized

OOS a \arn(A

gralt

Mrads,naiurnl

the sample

(Ii

yleld,immobilized \tructurr

It IS rlrar

lhat

capacity oi

starch

the sample

are

of two broken,so

I is a suitable

samples

are

release

of

support

ior

1042

Nguyen

Kclattonship

between

times

anh

residual

of repeated

et ul.

Dung

a( tivity

ot Immobilized

urease

and

USC’

References

1 Hoitman,A

S.(l Y81).A

processing

treatment

2 Kacjt\u

I j 1085).recrnt

I)olym~~r~atlon

polycJthvlc>n ‘i.Allnn,\ Schmer

and

review

ot the LI+ ot radiation

to prepare

novel

progres,

IO III(IC h(‘mil

Kndlat.Phys.Chem

27

therdpeutic

and

G.I)oancl,\Y

M ,Shasha

applic .t%.S.,and

mntrlx

In ( ontrolled

Release

Chemlc

al $0~ irty.\idshinglon.[)

of biocoml)onent

a/ u\(’

Knclint

I jcAnoyama,Liang

ot c’nzymt’s

didgnostic

and

biochemical

by rddiaiion

f’hys.Chem.Vol.L5,No.4-

i,! (- i2li

R.(;ombotz,\atorI

11 YHO).lmmobilization

chemical

Kadiat.f~hys.(~hem.Vol.lR,N~.l-

c,n thf. Immol~llization

the dl)ptlc-dtion

Hotfman,LZ/ayre

t)lomatrrlnl

plus

anrl

ation\.Kndiat Ku\sc.ll

PC\IIC I~(T (

I ‘I‘-

antibodies

C.Dong to grafted

dnd

Coltfried

polymers

ior

Phys.C~hcm.L7,265-273

c r ,l.nc nl)sulntlon ,A( 5 kvrn 74

of pesticides

$cAr 5 3 Sher.H.H..td..

within American

a starch