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J. Trop. Biodi . Biote h., Vol.

1 , —1

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The produ io of or ker el miso ased o ri e-koji fer e ted y
Aspergillus oryzae a d Rhizopus oligosporus
Diah Rat a i g u

*, Thel a Agusi a Budi ai , T i Da si i , Pa ji Cah a Ma a da

1

Resear h U it for Clea Te h ology, I do esia I situte of S ie es, Jala Cisitu/Sa gkuria g, Ba du g, 1 , I do esia
Depart e t of Che istry, Fa ulty of Mathe ai s a d Natural S ie es, U i ersity of Se elas Maret, Jl. Ir. Suta i No. A,
Je res, Surakarta, 1 , I do esia
*Correspo di g author: diah

@lipi.go.id
ARTICLE INFO

ABSTRACT

Ari le history:
Re ei ed / /
Re ei ed i e ised fo
A epted / /

Aspergillus oryzae a d Rhizopus oligosporus has

The suita ilit of o
/

/

ate ial to p odu e

o du ted o t o i po ta t fa to s i

a d salt o e t aio . The

Key ords:
Aspergillus oryzae
o ke el
iso
i e-koji
Rhizopus oligosporus
DOI: 1 . 1 /jt
© 1 JTBB

ke el as a

α-a

edia. The

a el

as p epa ed


old o posiio i

high dissol ed p otei

/

%,

ke el

iso

i to

ith high

%, a d

%


i tu e ate i e da s of fe

ee

e taio .

ith high glu ose a d

o e t aio . The esult sho ed that i e-koji o tai i g A. oryzae a d

R. oligosporus i the aio of : had the highest α-a
a d .

i e-koji

as opi ized to p odu e i e-koji

as also opi ized to p odu e o


as

i o ulai g the spo es of %

lase a d p otease a i it . Dife e t NaCl o e t aio s of

The salt o e t aio

i e-koji o tai i g

i tu e of oth i a aio of : , : , a d :

old o posiio

su je ted to opi izaio p o ess a d added to ea h

.

e ted


i esigated. The opi izaio

iso p odu io

old o posiio

A. oryzae, % R. oligosporus, a d % the
dife e t i e

iso fe

ee

lase a d p otease a i it of .

U/ L espe i el . I addiio , the p ese e of

% NaCl i

o


ke el

iso fe

U/ L
e ted

A. oryzae a d R. oligosporus i the aio of : e hi ited the highest glu ose a d dissol ed
p otei
iso

o e t aio of .6

g/ L a d .

e ted. The ut ie t

o te t a al sis sho ed


o

as fe

ke el

ed i to a

ut ie t e ha e e t ate

iso fo dieta

e ted so ea

e ted food o igi all f o

paste
Japa

a d has


ee

used

This food is apa le of edu i g a e
a i-dia ei ,

ultu e i

addiio

As a sta te

e taio .

as

sta te


e ted g ai

. Miso has

ell p olife aio a d

a i-h pe te si e

a io ida t p ope ies Hi ota et al.,

solid state fe

Bou di ho et al.,

se e al health e eits due to its ioa i e o pou ds su h
as pepides a d isola o es eleased du i g fe

ell

, .6


ke el ould e uilized to p odu e a o el o

ell as

asi i g edie t to ook soups as

, .

di e sii aio a d fo i p o i g ut iio al a d health status.

is a t adiio al

seaso i g fo side dishes Kusu oto a d Rai,

as

; K o et al.,

;

dete

i ed

hi h the koji
used fo

koji, a

Aspergilus oryzae se ed as
to so ea , salt, a d

ate

.
ultu e, koji has se e al t pes

the su st ates so ea , a le , o
olds g o . The

ost o

iso p odu io is i e-koji fe

hi h a ou ts
al.,

ade f o

% of

hi h a e
i e i

o t pe of koji
e ted

A. oryzae

iso p odu io i Japa

Ma hida et

. Ho e e , Aspergillus sojae, Aspergillus ta arii,

; Kusu oto

Aspergillus a a ori, Aspergillus saitoi, Aspergillus ka a hi,

. Miso has s eet, salt , a d u a i taste,

a d Rhizopus oligosporus ha e also ee used i Japa ese

)aid a d El She a
a d Rai,

o ludes o

e ted a d

eased 6.

o te t ha a te isi . This food is usuall

o ld ide as a

possesses

ke el

iso. Glu ose, dissol ed p otei , a d fat o te t i

. I trodu io
fe

ke el

% NaCl as su je ted to ut ie t
ke el as fe

i es espe i el . This stud

fe

ith

o te t a al sis a d o pa ed to the esult efo e the o
t a sfo

Miso

g/ L espe i el . The opi ized o

A. oryzae a d R. oligosporus i the aio of :

,

; Shi izu et al.,

p ese ts i a paste-like half-solid food fo

ith high p otei

food i dust ies fo the p odu io of koji Esaki et al., 99 ;

J. Trop. Biodi . Biote h., Vol.
Li

et al.,

e z

6 . The

koji p odu e h d ol i

es fo i sta e p oteases a d a

iologi al pol
o o e

the fe

u a i tastes of
a d

lases fo deg adi g

a ious e z

e taio

e taio

ai

ea io s

e a i iies a e opi ized

ield opi u

sta te

ultu es fo

a ufa tu e s Ma ui et al.,

Beside koji, the t pe of a

uilized to p odu e

to o e o e

Fa zo et al.,

iso ould suppo t dieta

ut iio

. Dieta

ed et al.,

a d health status issues

di e sit di e tl i lue es diet

; Mou si et al.,

de o st ated dieta

ould ofe a othe t pe of
O e of the

i o ut ie t de sit of
iso

ith a othe t pe of
ith dife e t ut ie t

iso is o

ke el. The

as

the p odu io of so ea

illio

as

. This highe a aila ilit

elia le fo

dieta

of o
ate

illio

odit

asis. Sta h a d p otei

oe
ut ie t
e taio

.6 % p otei ,

de t o

o tai ed

o

o ad
ke el a
o pou ds

e aluated

ith e z

a d

o e t aio

ke el a d i e

lo al sto e. Ha de , 9

a id TCA

Lo

Me k Da

. The a al i al g ade of Lo
Me k Da

stadt, Ge

i aio of dissol ed p otei
ethod Lo

, 9

a

a

eage t

a

a d used

o e t aio usi g

. The a al i al g ade of Nelso

eage ts a d a se o ol date
stadt, Ge

stadt, Ge

i aio of p otease a i it usi g Ku itz

as pu hased f o
fo dete

e e pu hased

. Casei a d T i hlo oa ei

e e pu hased f o

ethod Ku itz, 9

e e pu hased f o

a d used fo dete

o e t aio usi g Nelso -So og i

Me k

i aio of glu ose

ethod Nelso , 9

.

. . Methods
. . . Spores Suspe sio Preparaio
A. oryzae a d R. oligosphorus

e e i o ulated
o

sepa atel i to dife e t PDA a d i u ated at
da s. The spo es of ea h

ould

C fo se e

e e diluted i

. 9% NaCl,

a d the opi al de sit of ea h pu e ultu e as
6

easu ed at

usi g a spe t ophoto ete . The opi al de sit
e

as o i

dife e t PDA. The olo

u

o se ed a d ou ted ate
olo

fo

ed

as

CFU/ L.

plai g the

e of ea h

i to

ediu

as

o

h of i u aio at

ed is assu ed o i g f o

Ri e

as to o tai the

e taio opi izaio

esults

e a i it at the i e of fe

as also
e ted o

fo

Che ist , I do esia

C. O e

o e spo e.

. . . Koji fer e taio Pro ess

.

iso. The opi izaio

ee

e taio
of salt

o du ted a d e aluated

easu i g the o e t aio of glu ose a d dissol ed p otei
o ea h fe

I situte of S ie e. Co

ke el. Nut ie t o te t a al sis as

as lea sed,

ate i a aio of :
o

hi h a i p o e the ut iio al

ut iio al status of

Resea h U it fo

ee

e ted

o posiio of A. oryzae a d R. oligosporus ultu e aio, as
iso. The fe

o tai ed f o

The total ell u

He e, the pu pose of this stud

status of

to

di e sii aio

ello

that i p o es la o s, a o as, a d te tu es of fe

ell as NaCl o e t aio

iso fo dieta

set to . to o tai a ell o e t aio a ou d

ide di e sit of phe oli

food p odu ts Villege et al.,

ke el

oppo tu it

as ased o the a al sis

o e ted i to esse ial a i o a ids, si ple

a oh d ates, a d a

ould gi e a

Aspergillus oryzae a d Rhizopus oligosphorus

he eas

o

iso fe

a oh d ates,

6 . This data

ke el fo the o

de eased ate

. Mi roorga is s a d Che i als

, the

di e sii aio . Besides, the
%

eased o

. Materials a d Methods

et i to s

e de s

ke el is suita le fo

o sists of 6 .

e io he i all

Ju e

et i to s

.6

USDA,

% oil USDA,

i t odu e a o el o

as fe

o ld ide p odu io of

so ea . I

. 6

ut ie ts i

e taio . This stud

Da

ost pote ial g ai s to su situte a

is u e tl highe tha

o posiio of o

fe

the

a d used fo dete

o posiio a d i he e t health e eits.

p odu io of o

hethe

di e sit has st o g asso iaio

the diet. The efo e, su situi g so ea

.

e ted to k o

; a d Rah et al.,

ith ut iio al status, pa i ula l

hi h

ith the a al sis

ke el

e

gua a t i g ade uate i take of esse ial ut ie ts

ate ial of

as o pa ed

iso t pes. I addiio to

a d i po ta t o - ut ie t fa to s. Resea h o du ted

o

ut ie t o te t a al sis
o du ted efo e the o

iso. The use of othe g ai s as a

ate ial su situte fo p odu i g

g ai

the opi ized i al p odu t. The esult of

to i p o e ut iio al a d health status.

.

so ea , the e a e a lot of pote ial g ai s that ould

Ke

fat o te t i

ate ial is i po ta t to

di e sif the p odu io of dife e t

ualit

the a ou t of glu ose, dissol ed p otei , a d

p o ess ga e s eet a d

iso. These e z

o t olled to

di e sit

do e to k o

e s su h as p otei a d a oh d ate. The

p odu ed f o

happe ed i

fe

olds i

1 , —1

C fo

ashed, a d
/ . This

i utes a d ooled do

i ed

i tu e
to oo

ith disilled

as ste ilised at
te pe atu e.

The spo es of % A. oryzae, % R. oligosphorus, a d % the
i tu e of oth i a aio of : , : , a d :
i o ulated i to dife e t
i u ated at
fo

o

C fo

edia. Ea h

ee

ediu

as

h a d d ied i side the o e at

h. Ea h d ied ultu e

p o ess to o tai

i e

/

o

C

as the su je ted to a g i di g

i e-koji i the fo

of po de .
9

J. Trop. Biodi . Biote h., Vol.

1 , —1

. . . Cor Ker el Fer e taio
Co
ke el

ke el

as the

as

to oo

eight to gi e a

lea ed,
C fo

dife e t i e-koji o posiio
do e at
da s-fe

ke el

ke el

as

/

. Fe

C. To dea i ate the e z

as added to the

as ooled

te pe atu e fo

i ed

addi g

ith

e io ed a o e. Ea h i e-

as %

o

i at

g. Co

i utes. Ate it

te pe atu e, o

koji added to o

ass of

ashed, sli ed i to pie es, soaked,

o

a d ste ilised at
do

phosphate ufe pH .6 to asei soluio a d i u ated fo

e taio

as

o

C fo i e da s i side a i u ato . Du i g i e

L of disilled

Fe

e ted o

a d diluted to

p

fo 9

i the te pe atu e of

as

g as sa pled e e

e t ifuged at

p

Ea h supe ata t

C fo

i . The supe ata t

C.

usi g spe t ophoto ete . O e u it
as dei ed as the a ou t of e z
i at

e
o

C

L supe ata t.

/

ai a i it a al sis.

The soluio

o tai i g se e

. M phosphate ufe pH .
i . The sa ple soluio

the supe ata t as
sta h soluio

L of % solu le sta h

as p epa ed

ude e z
es

.6 N sulfu i a id a d .

L of

e e t a t to the solu le
i

as do e
L of

i tu e. The la k soluio

C fo

addi g

a d i u ated fo

Dea i aio of the e z

o

as i u ated at

o

at

addi g .

% sodiu

as p epa ed

C.

L of

of Salt Co e traio

Dife e t salt o e t aio s of

lase a d p otease

. . . α-A ylase Assay

the

i i the te pe atu e of

e e added to ea h fe
e taio . The

fe

e taio

fe

e taio , fe

fo eight

%

ke el ate i e

eeks

o e. Ate eight
ke el of

g

eeks of

as sa pled

L of phosphate ufe . The suspe sio

as shake at
p

%, a d

i tu es e e su je ted to fu the

e ted o

a d diluted to

%,

e ted o

p

fo 9

i the te pe atu e of

i a d e t ifuged at

o

C fo

i . The supe ata t

as olle ted a d su je ted to glu ose o e t aio a al sis
usi g Nelso -So og i

ethod So og i, 9

tu gstate to

p otei

o e t aio a al sis usi g Lo

addi g

al., 9

.

L

a d Che i al

A alysis

da s of fe

i

as
o

as olle ted a d su je ted to a so a e

. . . Opi izaio

i a d e t ifuged at

o

olle ted a d su je ted to α-a

e z

fo

a d e p essed as u its pe

da

as

L of phosphate ufe . The suspe sio

as shake at
p

ate to asei soluio a d

that eleases µg of a i o a id as t osi e fo

ke el of

oo

as p epa ed

t eated the sa e as sa ple soluio . Ea h soluio

of p otease a i it
. . . E zy ai A i ity A alysis

L of % TCA

i tu e a d i u ated at
i . The la k soluio

easu e e t at

e taio , the sa pli g p o ess as o du ted.

e, .

a d dissol ed
ethod Lo

et

of disilled ate to sta h soluio a d as t eated the sa e
as sa ple soluio . Ea h soluio
p

fo

. . . Nutrie t o te t a alysis

as e t ifuged at

i i the te pe atu e of

o

C. Ea h supe ata t

as olle ted a d su je ted to edu i g suga a al sis usi g
Foli Wu

ethod Foli a d Wu, 9 9 . Foli -Wu

ethod as

ased o the a ilit of edu i g suga s to edu e alkali e
oppe ta t ate. The p odu t of this ea io
o idise phospho ol di a id to

ield a

as used to

lue o pou d

easu ed usi g spe t ophoto ete at
as o pa ed

ith e z

ai a i it of sta da d α-a

o tai ed th ough the sa e
a i it

. The esult

ethod. O e u it of α-a

as dei ed as the a ou t of e z

ke el

ke el

o pa e the ut ie t efo e a d ate fe
Nelso -So og i
o e t aio

lase

ethod Ma i akiza et al.,

i

at

o

C a d

.

L of % asei

9

as p epa ed

supe ata t as

ude e z

a al sis usi g

, dissol ed p otei
ethod Lo

et al.,

o te t a al sis usi g so hlet e t a io
.

. . Physi al properies of koji a d Miso

ed

ased o

. The soluio

as i u ated at

sa ple soluio

e taio p o ess.

. Results a d Dis ussio s

o ego
Ku itz

ethod So og i, 9

a al sis usi g Lo

, a d fat

efo e the

as o du ted to

The a al sis i ludes glu ose o e t aio

9

. . . Protease Assay
ethod p oposed

as do e

e ted a d i the i al p odu t of o

iso that has ee opi ized. This

Koji
as pe fo

as fe

lase

L supe ata t.

The p otease assa

ut ie t o te t a al sis

e that eleases

µg of edu i g suga as glu ose fo
e p essed as u its pe

The
o

o tai i g

o

C fo

addi g

i . The

.

e e ta t a d

the

L of the
.

L of

ade f o
ith

i e that has

ould spo es

ee

east te peh a d the

i u ated fo th ee da s. Du i g koji fe
e z

es p oteases, a
oulds a d ha e a

o e hauli g

a

lases a d lipases
i po ta t ole i

ate ial

stea ed a d

e taio , a ious
ill e p odu ed
the p o ess of

a o ole ules i to si ple

ole ules Wah uhapsa i et al.,

; A da i et al.,

.

J. Trop. Biodi . Biote h., Vol.

Figure . The e z

The se o d stage of

ai a i it of α-a ilase i

iso p odu io is fe

i tu e of salt, pea uts a d koji to fo
is shaped like pea ut ute , o
ea

ello ish, light

e taio

ith a

iso la ou . Miso

olou ed pea ut ute

o

, da k

te tu e is like pea ut ute
a sa ou

a

i e-koji o tai i g dife e t

o

ith a

to la kish

ith

ith salt tasted. Miso p odu es

a d salt taste u i ue a d o

e i el

odate t o

ole ules of

The highest α-a

lase a i it

i tu e ga e α-a
6

old o posiio had dife e t a i it to h d ol ze sta h
fo

o

ke el Figu e

he e the a i it i
fou th da of fe
da . This

. All sa ples had si ila p oiles

eased f o

as possi l

aused

edia a d the staio a
edu io
fo
a

of

olds

aio of α-a
a fo

the i st da

de easi g ut ie t i the

phase egulaio
eta olis

lase e z

leadi g to the

ate. This est ai s the

e as it has ee thought to e

i oo ga is s to sa e e e g

a

lase a i it of .

o e t aio

99 . This esult
ho fou d i
the disso iaio

as o i

easi g

ed

the aio of

:

had highe α-a

o pa ed to the othe aios. Figu e
p odu ed α-a

lase e z

e

o e tha R. oligosporus. This

e at i uted to the fa t that the g o th of Rhizopus

spe ies i stea ed polished i e ultu e is dela ed due to
i a ilit of the
o tai ed i

i o es to uilize heat-de atu ed p otei s

i e Ta aka et al., 9

la k a o pepidase a i it
i apa le of

. Rhizopus st ai s also

hi h i

eaki g do

tu

as e t e el

Qia

et al.,

ould e ha e

The i di g a al sis sho s the ki ei a io of

e.

altose is

9 9 also
a d R.

eak i

i e a d

e a i it of koji o tai i g A.
the o es o tai i g R.

ith a highe aio. The esult sho ed du i g the

da s of fe

lo e α-a

e taio , the

lase a i it . This

g o th of ea h
phase fo

Ha a a a et al.

as ge e all highe tha

oligosporus
i st fe

F ede i k et al.

o sta t fo sta h i di g ith the e z

o du ted

so ea -koji. He e, the e z
oryzae

akes the

heat-de atu ed p otei s. I

oligosporus NRRL

e

lase a i it

also sho ed A. oryzae

lase a i it of R. oryzae IFO

e ha is

altose o e t aio

Takefuji et al.

epo ted the α-a

ould i hi it the a i it of h d olase e z

itself th ough p odu t i hi iio

o du ted

.

ho sho ed i e-koji o tai i g A. oryzae a d R.

oryzae i

a

U/ L Figu e

addiio , the stud

e. The p ese e of h d ol sates i a e tai

a

i e-koji

altose as o e of

lase h d ol sates is a le to hedge the p odu io of αlase e z

as sho

Co o ado et al.,

. Mo eo e , the e essi e a ou t of
α-a

u il the

e taio a d te d to di i ish o the ith

ole ule

i di g step is

o tai i g A. oryzae a d R. oligosporus i the aio of : .
This esult is si ila to the stud

lase a i it , the esult sho ed ea h

he

app o i atel 6. -fold eake .

. . The A i ity of α-A ylase E zy e
Rega di g α-a

altose. He e,

altose o e t aio is high a d a se o d

This

sau e, soup sau e a d soaked spi es fo pi kles.

i di g site is la ge e ough to

i d at the a i e site, the ai it of this

o l used as a

spi e o a a iet of Japa ese uisi e fo e a ple used as a

old o posiio .

o peii e. This

a o

1 , —1

old i the

adaptaio

i ed ultu e e hi ited

as p o a l

i ed ultu e

e ause the

as sill i the lag

p o ess leadi g to less α-a

lase

J. Trop. Biodi . Biote h., Vol.
e z

1 , —1

e p odu io . Mo eo e , i the

as o peiio
the

et ee

oth

olds to get ut ie t f o

edia. He e, A. oryzae a d R. oligosporus i the aio of

: a d : e hi ited lesse α-a

The opi izaio stud of

i ed ultu e, the e

lase a i it

o pa ed to

i e-koji o tai i g A. oryzae a d R. oligosporus i the aio of
:

e hi ited highe

Othe tha
ea h

α-a

esult sho ed all

of

i e-koji as also i esigated. The

old o posiio

a i it o the fou th da of fe
i di g o fo

Fu the

lase a i it , p otease a i it

old o posiio i

ea hed its

e taio

a i u

Figu e

. This

s to othe studies suggesi g that highest

p otease a i it of R. olisgosporus a d A. oryzae
o tai ed at

h of i u aio a d ate

; Ha et al.,

as usuall

de Cast o et al.,

. The p otease a i it of R. oligosporus

as highe tha A. oryzae, ho e e , the highest p otease
a i it

as sho

i e-koji o tai i g A. oryzae a d R.

oligosporus i the aio of : . This
a i it of .
sho

U/ L Figu e

i tu e ga e p otease

. The sa e esult

as also

Sta z ńska-Ja isze ska et al.

ho o se ed

the i p o e e t of p otei h d ol sis

he the dose of A.

oryzae spo es as e ual a d lo e tha that of R. oligosporus
i

i o ulu . Yigza

et al.

e ted

po a l

aused

he tef a d g ass-pea

A. oryzae a d R. oligosporus. This
the a ilit of A. oryzae to eli i ate

of p otease i hi ito s i so a ea i
p oteol sis
fe

he it is

e taio

i ed

Ho g et al.,

of α-a

lase a d p otease e z

o e, the data sho

a o e also i pl

o ultu i g A. oryzae a d R. oligosporus
p odu io of α-a
fe

lase a d p otease e z

e taio , these

olds olo ized o

es.
that

ould aise the
es. I solid-state

ke el as the

edia

th ough t o t pes of pe et ai e h phae. The lo g o es, also
alled s oui g h phae a e used to help the
o posiio of the
a oss the

ediu

old e plo e the

a d seek e

olo izaio sites

ediu . These h phae a e fo

ed du i g the lag

a d e po e ial phase
a ou t of O i the

he e the e is sill a sig ii a t

ediu . Mea

hile, the sho t h phae,

also alled egetai e h phae a e used to a ho the olo
to the solid
olo

ediu

p e e i g the e o al of the e i e

due to ph si al distu a es like i d a d agitaio of

the fe

e taio

ed. These h phae a e used to i p o e

su st ate h d ol sis
e z

es i to the

se ei g la ge ua iies of h d ol i

ediu

Sugai-Gu ios et al.,

6.

also dis o e ed the

i p o e e t of a i o a id p oile
e e fe

lase a d p otease a i iies

e s of A. oryzae a d R. oligosporus ould o t ol

the p odu io
. . The A i ity of Protease E zy e

α-a

o pa ed to the othe s. This esult suggested o t olli g the
spo e u

ala e aio of : .

old o posiio sho ed

ost

easi g the deg ee of

ith R. oligosporus du i g the
.

. . The Opi izaio of Salt Co e traio
Salt is o e of

as

o po e ts to p odu e

iso hi h

also eeds to e opi ized. Dife e t o e t aio s of salt
e e added to the

i tu e i

o e t aio p odu ed highe
p otei as i di ato s fo
o du ted

Figure . The p otease a i it i

ai

i e-koji o tai i g dife e t

Ito et al.

old o posiio

o de to k o

hi h salt

o e t aio of glu ose a d

ete fe

e taio

esult. The stud

e ealed a i te esi g fa t

J. Trop. Biodi . Biote h., Vol.
that the salt

o tai ed i

p essu e a d

as

iso did

ot asso iated

hu a s. Ho e e , the easo

ot i

ease

lood

ith h pe te sio

ke el, the o e fe

of

hlo ide io s to

edu ed

he

fe

e ted

he

% of NaCl

e ted

o e t aio de eased

highest glu ose o e t aio of .6

o e t aio

highest α-a

. This

g/ L o pa ed to

old o posiio

e a i e. He e, the

had the

lase a i it as des i ed a o e leadi g to the

as applied i

o

as

ke el

iso

R. oligosporus. Ho e e , the glu ose

A. oryzae: R. oligosporus i the aio of : p odu ed the
the othe s see Figu e

e ui es a highe

p odu io of glu ose as o e of the h d ol sis p odu ts

e ai s u lea a d the stud

% of NaCl as added to o

R. oligosporus p o a l

o e t aio

i

sill eeds to e o du ted fu the . The esult sho ed

possessed

1 , —1

he

as added. High

of salt ould also i hi it the g o th of the

olds espe iall if oth
The efo e,

% of NaCl

he

olds a e ot halophile

% NaCl

p odu io

glu ose i

li ited leadi g to lo e p odu io of glu ose. This i di g

ke el

iso fe

de eased as salt o e t aio
sho ed the opi u
o posiio

salt

e ted
i

those

eased. The esult also

o e t aio

as dife e t. Whe

olds

o

fo

ke el

R. oligosporus the sa ple ea hed its
o e t aio i the p ese e of

ea h
as fe

as i li e

glu ose

% NaCl. A si ila

esult

old

ea h o

ke el

o posiio

6; Fe tahu a d Ku a i,

.

o e t aio of

iso i dife e t salt o e t aio

easu ed. The esult sho ed o

as

epo ts Al a sou i et al.,

I addiio to glu ose, the p otei

e ted

a i u

ea h

ith so e s ie ii

; Duta et al.,

old

lase i

i o es.
i tu e, the

highest glu ose o e t aio . Ho e e , the o e t aio of
o

of α-a

as added to the

ke el

iso fe

as also
e ted

as sho

he A. oryzae a d R. oligosporus i the aio of

A. oryzae: R. oligosporus i the aio of : p odu ed the

: a d :

as used to fe

highest dissol ed p otei

glu ose o e t aio
as added to the

e t o

ke el. The

as also ea hed

he

a i u

ke el

iso had the

lo est o e t aio of glu ose i the p ese e of
e ept fo R. oligosporus
he

% NaCl

as added to the
as p o a l

o e t aio of hlo ide io i the

o e of the alloste i a i ato s fo a
a d Kha e,

the

% of NaCl hi h is
lase e z

e Ku a

; Mohapat a et al., 99 ; O o a d Ajele,

esidues a d uli atel tu

i tu e Figu e

. This

o ฀-a

; Willia s et al.,

ith atal i

lase a i it

6 . The a

Mau us et

lase e z

Figure . The efe t of a ious salt o e t aio
o e t aio i o ke el Miso

had the highest p otease a i it . Mo eo e , all
o posiio s i
p otei

this stud

e

ea hed

a i u

o e t aio i the p ese e of

NaCl added to the

i tu e. I

aused

99 . Chlo ide io s ould i d a d i te a t
al.,

% NaCl

hi h had the lo est glu ose

the ase of R. oligosporus, this
lo

as added to the

g/ L

he
as

e ause a o di g to the p e ious esult, this o posiio

i tu e.

The esult also sho ed all o

o e t aio

% of NaCl

% of NaCl

o e t aio of .

o e t aio
de atu aio

dissol ed

% NaCl. The

i tu e, the lo e

old
oe

the p otei

dete ted due to sali g out a d p otease
Su et al.,

i hi it the g o th of

. High salt o e t aio s also
a

i oo ga is s,

hi h i tu

g eatl de eases the likelihood of the p otease e z

es

p odu io . These a e espo si le fo p otei h d ol sis to
fo

a i o a ids. The efo e, the esult sho ed

o e t aio i

eased to

%a d

he salt

%, dissol ed p otei i

ith aio o pa iso of A oryzae da R. oligosporus o glu ose

J. Trop. Biodi . Biote h., Vol.

1 , —1

Figure . The I te a io et ee a ious of salt o e t aio a d o posiio A. oryzae a d R. oligosporus o
p otei o e t aio i o ke el Miso

o

ke el

iso de eased. This i di g

as si ila to so e

epo ted a i les sa i g p oteol i a i it de eased as salt
o e t aio i

eased Dodia et al.,

; Patel et al.,
opi izaio
ke el

6; Ga di i et al.,

; Ku ia a et al

. Based o the

esults a o e, i the p ese e of

iso fe

e ted

to

edu e

A. oryzae a d R. oligosporus i the

Yigza et al.,

of

the

eu oto i

a id β-ODAP i

fe

fo

iso fe

e ted

oryzae a d R. oligosporus i the aio of :
ke el

easu ed

efo e it

as fe

i di ato s

ith

e ted. The e

i ludi g

i

i e

hethe

eased o de eased ate fe

that all
The
i

easu ed i di ato s i

o e t aio
eased 6.

6.

g/ L Ta le

e e th ee
p otei

the

ut ie t

o te t

e taio . The esult sho ed
eased ate fe
i

e taio f o

. Dissol ed p otei of o

e taio .
o

.

ke el
g/ L to

ke el

iso

as o e t ated due to ate loss i the d i g p o ess a d
i

eased due to p otease a i it p odu ed

R. oligosporus. Mea
i

eased .

a

lase a i it

hile the glu ose

i es f o
those

.

hi h i tu

olaile o pou ds fo spe ii a o as Pei ado et al.,

Sa ples
Co ke el efo e Opi ized o
fe e taio
ke el Miso

Dissol ed p otei
g/ L
Glu ose g/ L

% NaCl

dissol ed

of dissol ed p otei

i es ate fe

i e the a o a of

Ta le . Nut iio al o te t A al sis of o
ke el Miso
fe e ted A. oryzae: R. oligosporus i the aio of :

A.

o e t aio , glu ose o e t aio , a d fat o te t. This
as do e to dete

hi h aids the

fat a ids a d gl e ol. Fat

6.

g/ L to . 9

A. oryzae a d
o e t aio
g/ L due to

olds du i g the fe

e taio

.

± .

6.

. 6± .

Fat Co te t %

as su je ted to ut ie t o te t a al sis a d o pa ed to
o

R. oligosporus a d A. oryzae

I di ato

ke el

eased

% due to lipase a i it

iso si e fat a ids a e p o e to o idaio

old o posiio a d salt o e t aio ha e
o

.

h d ol sis of t igl e ide to fo

. . Nutrie t Co te t A alysis
O e

.9 % to

iso also i

β-N-o al l-α,βe ted g ass pea

.

ee opi ized, the hose

i es f o

p odu ed

old o posiio has also ee p o e

%

dia i op opio i

.6

ke el

o te t is a i po ta t fa to to dete

% NaCl, o

aio of : p odu ed highest glu ose a d dissol ed p otei
o e t aio . This

p o ess. The fat o te t of o

.

.9 ± .

± .
± .

.

± .

. Co lusio
The o

ke el e plo ed i this stud

e used as a
o

i aio

a

ate ial fo

et ee

as suita le to

iso p odu io . The

A. oryzae a d R. oligosporus i

aio of : i side the i e-koji ould ield highest α-a
a d p otease a i it

of

espe i el . The addiio
fe

e taio

of

U/ L a d

.

lase
U/ L

% NaCl i lue ed

ete

esults as the glu ose a d dissol ed p otei

o e t aio s
fe

.

the

e e the highest

e ted

easu ed i

the

iso

the opi ised

i e-koji. The glu ose a d

dissol ed p otei

o e t aio

easu ed i the opi ised

p odu t as .6

g/ L a d .

g/ L espe i el . These

data i pl i

eased o e t aio of glu ose, p otei , α-

J. Trop. Biodi . Biote h., Vol.
a

lase a i it , a d p otease a i it

o ultu i g

ould

o posiio

Duta, T. K., Ja a, M., PAHARI, P. R., & Bhata ha a, T.,

ould e o tai ed
a d

o t olli g

The efe t of te pe atu e, pH, a d salt o a

salt

Heliodiapto us iduus Gu e

o e t aio . The ut ie t o te t e ha ed as sho
glu ose, dissol ed p otei , a d fat o te t i
iso as highe tha

o

o

ke el efo e it as fe

e ted. To

p odu io

fo

o

ke el. Fu the

A io ida t a i it
fe

e

ke el

e ted

as suppo ted

6,

.

-

food

a d

diets

usi g

,

agri ultural

Biodi e sit I te aio al.

I do esia I situte of

S ie es. The autho s ould like to tha k to Chelsie Ke
UK Defe se S ie e a d Te h olog La o ato

Fe tahu , M., & Ku a i, P. V.,

ed

of a

a d Ti

e ts a d suggesio s f o

thei a

a li guisi poi t

ophili

i g
of

lase a i it , Afri a Jour al of Mi ro iology
,

-

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Foli , O., & Wu, H., 9 9, A s ste
Refere es

spo e fo

sta h i h soil a d ha a te izaio

Resear h

of ie .

, Isolaio a d s ee i g

lase p odu i g the

Ba illi f o

NYC Oi e of the Chief Medi al E a i e fo

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so ea s

iodi ersity to i pro e utriio a d health. Ne Yo k:

ok

O B ie f o

fo

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Fa zo, J., Hu te , D., Bo elli, T., & Matei, F.,

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a d Food Che istry

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the o

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Esaki, H., O ozaki, H., Ka akishi, S., & Osa a, T.,

iso

esea h should

6,

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