I
nte
rna
t
io
na
l J
o
urna
l o
f
Adv
a
nces in Applie
d Science
s
(
I
J
AAS)
Vo
l.
1
5
,
No
.
1
,
Ma
r
ch
20
2
6
,
p
p
.
2
2
7
~
2
3
5
I
SS
N:
2252
-
8
8
1
4
,
DOI
:
1
0
.
1
1
5
9
1
/ijaas
.
v
1
5
.
i
1
.
pp
227
-
2
3
5
227
J
o
ur
na
l ho
m
ep
a
g
e
:
h
ttp
:
//ij
a
a
s
.
ia
esco
r
e.
co
m
Tem
pera
ture
and
pH
ef
fect
s
on bio
ethano
l produc
tion
from
wild cas
sa
v
a
(
Ma
niho
t gla
zio
vii
M
uell.
Arg
)
using
si
multa
neo
us
co
-
fermen
tatio
n
I
da
Ay
u P
rida
ri
T
a
ntr
i
1
,
I
da
B
a
g
us
Wa
y
a
n G
un
a
m
1,
2
,
An
a
k
Ag
un
g
M
a
de
Dew
i A
ng
g
re
ni
1,
2
,
I
G
ede
Ary
a
Su
j
a
na
2
1
D
e
p
a
r
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me
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o
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e
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a
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r
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d
a
y
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n
a
U
n
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si
t
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B
a
d
u
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g
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I
n
d
o
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s
i
a
2
La
b
o
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t
o
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o
f
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i
o
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n
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u
s
t
r
y
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a
c
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l
t
y
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g
r
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c
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l
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r
a
l
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c
h
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d
a
y
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n
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t
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B
a
d
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g
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I
n
d
o
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s
i
a
Art
icle
I
nfo
AB
S
T
RAC
T
A
r
ticle
his
to
r
y:
R
ec
eiv
ed
No
v
2
6
,
2
0
2
5
R
ev
is
ed
Dec
1
0
,
2
0
2
5
Acc
ep
ted
J
an
1
,
2
0
2
6
Bio
e
th
a
n
o
l
is
a
c
lea
n
a
lt
e
rn
a
ti
v
e
e
n
e
rg
y
s
o
u
rc
e
,
wi
th
wi
ld
c
a
ss
a
v
a
(
M
a
n
i
h
o
t
g
la
zi
o
v
ii
M
u
e
ll
.
Arg
)
a
s a
p
o
ten
ti
a
l
fe
e
d
sto
c
k
.
F
e
rm
e
n
tatio
n
c
o
n
v
e
r
ts g
lu
c
o
se
fro
m
h
y
d
r
o
ly
sis
in
t
o
e
t
h
a
n
o
l.
T
h
is
stu
d
y
e
x
a
m
in
e
s
t
h
e
e
ffe
c
ts
o
f
p
H
a
n
d
fe
rm
e
n
tatio
n
tem
p
e
ra
tu
re
o
n
b
io
e
th
a
n
o
l
c
h
a
ra
c
teristics
u
sin
g
a
sim
u
lt
a
n
e
o
u
s
sa
c
c
h
a
rifi
c
a
ti
o
n
a
n
d
c
o
-
fe
rm
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n
tatio
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(
S
S
CF
)
tec
h
n
i
q
u
e
.
A
fa
c
to
rial
ra
n
d
o
m
ize
d
b
lo
c
k
d
e
sig
n
(RBD)
wa
s
u
se
d
with
two
fa
c
to
rs:
p
H
(4
.
5
,
5
.
0
,
a
n
d
5
.
5
)
a
n
d
fe
rm
e
n
tatio
n
tem
p
e
ra
tu
re
(3
0
,
3
2
.
5
,
a
n
d
35
°C
).
Da
ta
we
r
e
a
n
a
ly
z
e
d
u
sin
g
v
a
rian
c
e
a
n
d
D
u
n
c
a
n
’s
tes
t.
Re
su
lt
s
sh
o
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d
t
h
a
t
p
H
a
n
d
tem
p
e
ra
tu
re
sig
n
ifi
c
a
n
t
ly
a
ffe
c
ted
p
H
v
a
lu
e
,
t
o
tal
so
lu
b
le
so
li
d
s,
re
d
u
c
in
g
su
g
a
r,
a
n
d
e
th
a
n
o
l
c
o
n
ten
t
.
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e
o
p
ti
m
a
l
c
o
n
d
it
i
o
n
s
f
o
r
b
i
o
e
th
a
n
o
l
p
ro
d
u
c
ti
o
n
we
re
p
H
4
.
5
a
n
d
tem
p
e
ra
tu
re
3
2
.
5
°C
,
y
ield
i
n
g
a
p
H
o
f
3
.
5
5
±
0
.
0
7
,
to
tal
so
lu
b
le
so
li
d
s
o
f
9
.
3
±0
.
5
7
°Brix
,
re
d
u
c
in
g
su
g
a
r
o
f
3
.
0
3
8
±0
.
1
0
m
g
/mL,
a
n
d
e
th
a
n
o
l
c
o
n
ten
t
o
f
3
.
4
8
±
0
.
3
7
(%
w/v
).
Ba
se
d
o
n
t
h
e
re
su
lt
s
o
f
th
is stu
d
y
,
wil
d
c
a
ss
a
v
a
c
a
n
b
e
u
ti
li
z
e
d
a
s
b
io
e
th
a
n
o
l
b
y
c
o
n
sid
e
ri
n
g
th
e
e
ffe
c
t
o
f
fe
rm
e
n
tatio
n
c
o
n
d
i
ti
o
n
s.
K
ey
w
o
r
d
s
:
B
io
eth
an
o
l
pH
S
im
u
ltan
eo
u
s
c
o
-
f
e
r
m
en
tatio
n
s
ac
ch
ar
if
icatio
n
T
em
p
er
atu
r
e
W
ild
c
ass
av
a
T
h
is i
s
a
n
o
p
e
n
a
c
c
e
ss
a
rticle
u
n
d
e
r th
e
CC B
Y
-
SA
li
c
e
n
se
.
C
o
r
r
e
s
p
o
nd
ing
A
uth
o
r
:
I
d
a
B
ag
u
s
W
ay
an
Gu
n
am
Dep
ar
tm
en
t o
f
Ag
r
o
in
d
u
s
tr
ial
T
ec
h
n
o
lo
g
y
,
Facu
lty
o
f
Ag
r
icu
ltu
r
al
T
ec
h
n
o
lo
g
y
,
Ud
ay
a
n
a
Un
iv
er
s
ity
B
u
k
it Ji
m
b
ar
an
,
B
ad
u
n
g
B
ali
-
8
0
3
6
1
,
I
n
d
o
n
esia
E
m
ail:
ib
wg
u
n
am
@
u
n
u
d
.
ac
.
id
1.
I
NT
RO
D
UCT
I
O
N
E
n
er
g
y
h
as
em
er
g
e
d
as
an
im
p
o
r
tan
t
is
s
u
e
r
ec
e
n
tly
i
n
th
e
wo
r
ld
,
af
f
ec
tin
g
th
e
ec
o
n
o
m
ic
d
ev
elo
p
m
e
n
t
alo
n
g
with
th
e
g
r
o
wth
o
f
p
o
p
u
latio
n
,
f
u
el
e
m
is
s
io
n
p
r
o
b
lem
s
,
an
d
n
o
t
to
m
en
tio
n
d
ep
letio
n
o
f
o
il
r
eser
v
es.
T
h
e
r
elen
tles
s
ly
in
cr
ea
s
in
g
co
n
s
u
m
p
tio
n
o
f
f
o
s
s
il
en
er
g
y
h
as
tr
ig
g
er
ed
t
h
e
cr
is
is
o
f
f
u
el
[
1
]
.
T
h
e
s
ca
r
city
an
d
im
p
ac
t
o
f
en
v
i
r
o
n
m
en
tal
p
o
llu
tio
n
d
u
e
to
th
e
u
s
e
o
f
f
o
s
s
il
f
u
els
h
as
en
co
u
r
a
g
ed
m
an
y
in
n
o
v
atio
n
s
to
f
in
d
r
en
ewa
b
le
an
d
en
v
ir
o
n
m
en
tally
f
r
ien
d
ly
alter
n
ativ
e
f
u
els.
T
h
e
s
o
lu
tio
n
to
en
er
g
y
p
r
o
b
lem
s
a
n
d
t
h
e
g
r
ee
n
h
o
u
s
e
ef
f
ec
t
ca
u
s
ed
b
y
th
e
u
s
e
o
f
f
o
s
s
il
f
u
els
is
th
e
d
ev
elo
p
m
en
t
o
f
alter
n
ativ
e
f
u
els
.
On
e
o
f
th
ese
alter
n
ativ
e
f
u
els
is
b
io
eth
an
o
l
[
2
]
.
B
io
eth
an
o
l
i
s
an
alter
n
ativ
e
th
at
will
b
e
n
ee
d
ed
in
th
e
f
u
tu
r
e
,
an
d
its
p
r
o
d
u
ctio
n
will
in
cr
e
ase
s
ig
n
if
ican
tly
b
e
ca
u
s
e
o
f
th
e
lar
g
e
n
u
m
b
er
o
f
r
aw
m
a
ter
ials
av
ailab
le
to
p
r
o
d
u
ce
b
io
eth
a
n
o
l
[
3
]
.
B
i
o
e
t
h
a
n
o
l
p
r
o
d
u
c
t
i
o
n
as
a
n
alt
e
r
n
a
t
i
v
e
e
n
e
r
g
y
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o
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r
c
e
i
s
n
o
t
h
i
n
g
n
e
w
.
B
i
o
e
t
h
a
n
o
l
p
r
o
d
u
c
t
i
o
n
c
u
r
r
e
n
t
l
y
s
t
il
l
u
s
es
f
o
o
d
r
a
w
m
a
t
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i
a
l
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n
e
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e
s
s
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f
o
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u
m
a
n
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f
e
,
s
u
c
h
a
s
c
o
r
n
,
b
e
a
n
s
,
a
n
d
t
u
b
e
r
s
s
u
c
h
a
s
c
a
s
s
a
v
a
[
4
]
.
B
i
o
e
t
h
a
n
o
l
p
r
o
d
u
c
t
i
o
n
f
r
o
m
t
h
i
s
m
a
t
e
r
i
al
c
a
n
s
o
l
v
e
e
n
e
r
g
y
p
r
o
b
l
e
m
s
,
b
u
t
i
f
i
m
p
l
e
m
e
n
t
e
d
o
n
a
l
a
r
g
e
s
c
a
l
e
,
i
t
w
il
l
c
r
e
a
t
e
n
e
w
p
r
o
b
l
e
m
s
,
s
u
c
h
as
f
o
o
d
s
h
o
r
t
a
g
e
s
f
o
r
h
u
m
a
n
l
i
f
e
.
O
n
e
o
f
t
h
e
i
n
e
d
i
b
l
e
n
a
t
u
r
al
m
a
t
e
r
i
a
ls
t
h
at
h
as
t
h
e
p
o
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e
n
t
i
a
l
t
o
b
e
u
s
e
d
a
s
a
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a
w
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a
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i
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f
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a
k
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b
i
o
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t
h
a
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l
i
s
w
il
d
c
a
s
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a
v
a
(
M
a
n
i
h
o
t
g
la
z
i
o
v
i
i
M
u
e
ll
.
Arg
)
.
Evaluation Warning : The document was created with Spire.PDF for Python.
I
SS
N
:
2
2
5
2
-
8
8
1
4
I
n
t J Ad
v
Ap
p
l Sci
,
Vo
l.
1
5
,
No
.
1
,
Ma
r
c
h
2
0
2
6
:
227
-
2
3
5
228
W
i
l
d
c
a
s
s
a
v
a
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s
a
n
o
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-
f
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o
d
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d
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h
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l
l
y
s
o
l
d
[
5
]
.
W
il
d
c
a
s
s
a
v
a
i
s
m
o
s
t
l
y
c
o
m
p
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d
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f
s
t
a
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w
h
i
c
h
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s
a
r
o
u
n
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7
8
%
o
f
t
h
e
d
r
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w
e
i
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t
.
O
t
h
e
r
c
o
m
p
o
n
e
n
t
s
o
f
w
i
l
d
ca
s
s
a
v
a
i
n
c
l
u
d
e
wa
t
e
r
,
p
r
o
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e
i
n
,
f
a
t
,
f
i
b
e
r
,
a
n
d
a
s
h
.
W
i
l
d
c
as
s
a
v
a
f
i
b
e
r
c
o
n
s
is
ts
o
f
c
e
l
l
u
l
o
s
e
,
h
e
m
ic
e
l
l
u
l
o
s
e
,
l
i
g
n
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n
,
a
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d
p
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n
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[
6
]
.
T
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y
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e
l
d
o
f
w
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c
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a
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u
c
ti
o
n
p
r
o
c
e
s
s
i
s
v
e
r
y
d
e
p
e
n
d
e
n
t
o
n
t
h
e
a
b
i
l
i
t
y
o
f
t
h
e
p
r
o
c
e
s
s
t
o
h
y
d
r
o
l
y
z
e
w
i
l
d
ca
s
s
a
v
a
c
o
m
p
o
n
e
n
t
s
,
e
s
p
e
ci
a
ll
y
s
t
a
r
c
h
i
n
t
o
g
l
u
c
o
s
e.
T
h
e
h
i
g
h
o
r
l
o
w
c
o
n
t
e
n
t
o
f
h
y
d
r
o
l
y
z
e
d
g
l
u
c
o
s
e
wi
l
l
i
n
f
l
u
e
n
c
e
t
h
e
b
i
o
et
h
a
n
o
l
f
o
r
m
a
ti
o
n
p
r
o
c
e
s
s
i
n
t
h
e
f
e
r
m
e
n
t
a
t
i
o
n
p
r
o
c
e
s
s
[
6
]
.
B
io
eth
an
o
l
in
th
is
s
tu
d
y
was
p
r
o
d
u
ce
d
u
s
in
g
s
ev
er
al
m
ai
n
p
r
o
ce
s
s
s
tag
es,
n
am
ely
g
el
atin
izatio
n
,
s
im
u
ltan
eo
u
s
s
ac
ch
ar
if
icatio
n
an
d
co
-
f
er
m
en
tatio
n
(
SS
C
F)
,
an
d
d
is
till
atio
n
.
T
h
e
en
zy
m
atic
h
y
d
r
o
l
y
s
is
p
r
o
ce
s
s
ca
n
b
e
ca
r
r
ie
d
o
u
t
s
im
u
ltan
eo
u
s
ly
with
co
-
f
e
r
m
en
t
atio
n
o
f
g
lu
co
s
e,
ce
llu
lo
s
e,
a
n
d
x
y
lo
s
e
,
wh
ich
is
ca
lled
SS
C
F
[
7
]
.
T
h
e
ad
v
a
n
ta
g
e
o
f
SS
C
F
is
th
at
p
r
o
d
u
ctio
n
co
s
ts
ca
n
b
e
r
e
d
u
ce
d
,
th
e
en
zy
m
atic
h
y
d
r
o
ly
s
is
p
r
o
ce
s
s
ca
n
b
e
elim
in
ated
,
a
n
d
th
e
p
r
o
d
u
ctiv
ity
o
f
eth
an
o
l
p
r
o
d
u
ctio
n
will
b
e
h
ig
h
er
th
an
th
e
s
ep
ar
ate
h
y
d
r
o
ly
s
is
an
d
f
er
m
en
tatio
n
p
r
o
ce
s
s
es
[
8
]
.
C
o
-
f
er
m
e
n
tatio
n
r
eq
u
ir
es
an
e
n
g
in
e
er
in
g
p
r
o
ce
s
s
o
f
m
icr
o
o
r
g
an
is
m
s
to
cr
ea
te
o
p
tim
al
co
n
d
itio
n
s
d
u
r
in
g
f
er
m
en
tatio
n
,
th
er
eb
y
in
cr
ea
s
in
g
eth
an
o
l
y
ield
.
T
h
e
SS
C
F
p
r
o
ce
s
s
tech
n
o
l
o
g
y
is
a
s
in
g
le
f
e
r
m
en
ter
f
er
m
e
n
tatio
n
p
r
o
ce
s
s
th
at
ca
n
s
h
o
r
ten
p
r
o
d
u
ctio
n
tim
e
an
d
r
ed
u
ce
p
r
o
d
u
ctio
n
co
s
ts
with
o
u
t
th
e
ad
d
itio
n
o
f
p
u
r
e
en
zy
m
es
f
o
r
h
y
d
r
o
ly
s
is
.
T
h
e
SS
C
F
p
r
o
ce
s
s
ca
n
b
e
u
s
ed
to
p
r
o
d
u
ce
b
io
eth
a
n
o
l
f
r
o
m
wild
ca
s
s
av
a
s
tar
ch
u
s
in
g
m
i
cr
o
o
r
g
an
is
m
s
th
at
p
r
o
d
u
ce
e
x
tr
ac
ellu
lar
en
zy
m
es.
E
n
zy
m
e
ac
tiv
ity
is
in
f
lu
en
ce
d
b
y
s
ev
er
al
f
ac
to
r
s
,
n
a
m
ely
te
m
p
er
atu
r
e,
p
H,
s
u
b
s
tr
ate
co
n
c
en
tr
atio
n
,
en
z
y
m
e
co
n
ce
n
tr
atio
n
,
a
n
d
t
h
e
p
r
esen
ce
o
f
in
h
ib
ito
r
s
[
6
]
.
Acc
o
r
d
in
g
to
Salih
u
et
a
l.
[
9
]
,
tem
p
e
r
a
tu
r
e
an
d
p
H
ar
e
th
e
m
o
s
t im
p
o
r
tan
t f
ac
t
o
r
s
to
b
e
k
n
o
wn
in
th
e
f
er
m
e
n
tatio
n
p
r
o
c
ess
b
ec
au
s
e
ea
ch
en
zy
m
e
f
u
n
c
tio
n
s
o
p
tim
ally
at
a
ce
r
tain
tem
p
er
atu
r
e
an
d
p
H.
Ab
o
v
e
t
h
e
o
p
tim
al
tem
p
e
r
atu
r
e,
th
e
r
ea
ctio
n
r
ate
d
ec
r
ea
s
e
s
s
h
ar
p
ly
b
ec
au
s
e
en
zy
m
es a
r
e
p
r
o
tein
s
th
at
d
e
n
atu
r
e
at
h
ig
h
tem
p
er
atu
r
es.
Sev
er
al
s
tu
d
ies
o
n
b
io
et
h
an
o
l
f
r
o
m
wild
ca
s
s
av
a
u
s
in
g
th
e
s
im
u
ltan
e
o
u
s
s
ac
ch
ar
if
i
ca
tio
n
an
d
f
er
m
en
tatio
n
(
SSF
)
an
d
SS
C
F
m
eth
o
d
s
r
ep
o
r
ted
th
at
th
e
SS
F
p
r
o
ce
s
s
u
s
in
g
ca
s
s
av
a
s
u
b
s
tr
ate
with
th
e
h
elp
o
f
th
e
m
icr
o
o
r
g
a
n
is
m
s
A
s
p
erg
ill
u
s
n
ig
er
an
d
S
a
cc
h
a
r
o
myc
es
ce
r
ev
is
ia
e
ca
n
p
r
o
d
u
ce
u
p
to
8
%
b
io
eth
an
o
l
u
s
in
g
a
r
aw
ca
s
s
av
a
s
u
b
s
tr
ate
with
a
p
H
o
f
5
.
0
an
d
f
er
m
en
tatio
n
tem
p
er
at
u
r
e
3
5
°C
p
r
o
d
u
ce
d
eth
an
o
l
lev
els
o
f
4
.
3
0
g
/1
0
0
m
L
[
1
0
]
.
Oth
er
r
esear
ch
er
s
h
av
e
r
ep
o
r
te
d
th
at
b
io
eth
an
o
l
ca
n
b
e
p
r
o
d
u
ce
d
f
r
o
m
r
aw
ca
s
s
av
a
b
y
co
-
cu
ltiv
atin
g
1
0
%
A
s
p
erg
illu
s
n
ig
er
,
R
h
iz
o
p
u
s
o
lig
o
s
p
o
r
u
s
,
Tr
ich
o
d
erma
r
ee
s
ei
,
a
n
d
S
a
cc
h
a
r
o
myc
es
ce
r
ev
is
ia
e
with
a
p
H
o
f
5
.
0
an
d
a
f
er
m
en
tatio
n
tem
p
er
at
u
r
e
o
f
3
0
°C
to
p
r
o
d
u
ce
an
eth
an
o
l
co
n
ten
t
o
f
1
2
.
9
g
/L
[
1
1
]
.
T
h
e
SS
C
F
p
r
o
ce
s
s
u
s
es
r
ice
s
tr
aw
as
a
s
u
b
s
tr
ate
with
th
e
h
elp
o
f
Sacc
h
ar
o
m
y
ce
s
ce
r
e
v
is
iae
m
icr
o
o
r
g
an
is
m
s
an
d
x
y
la
n
a
s
e
en
zy
m
es
with
a
p
H
o
f
4
.
8
a
n
d
a
tem
p
er
atu
r
e
o
f
34
°C
,
wh
i
ch
ca
n
p
r
o
d
u
ce
an
eth
an
o
l
co
n
ten
t
o
f
0
.
3
5
w/w
[
1
2
]
.
R
ec
en
tly
,
Gu
n
am
et
a
l.
[
6
]
r
ep
o
r
ted
th
at
th
e
h
ig
h
est
eth
an
o
l
y
ield
(
2
8
.
2
7
7
±
0
.
2
2
8
g
/L)
was
o
b
tain
ed
f
r
o
m
th
e
wild
ca
s
s
av
a
f
lo
u
r
(
W
C
F)
s
u
b
s
tr
ate
f
er
m
e
n
ted
u
s
in
g
a
co
-
c
u
ltu
r
e
o
f
R
5
I
4
is
o
late,
A
s
p
erg
illu
s
n
i
g
er
,
an
d
R
5
I
3
is
o
late
af
ter
8
d
ay
s
o
f
in
cu
b
atio
n
.
I
n
t
h
i
s
r
e
s
e
a
r
c
h
,
t
h
e
b
i
o
e
t
h
a
n
o
l
p
r
o
d
u
c
t
i
o
n
p
r
o
c
e
s
s
w
a
s
c
a
r
r
ie
d
o
u
t
u
s
i
n
g
t
h
e
S
S
C
F
t
e
c
h
n
iq
u
e
u
s
i
n
g
l
o
c
a
l
m
ic
r
o
b
i
a
ls
C
a
n
d
i
d
a
o
r
t
h
o
p
s
i
l
o
s
i
s
R
5
I
3
a
n
d
K
l
e
b
s
ie
l
l
a
va
r
i
i
c
o
l
a
R
5
I
4
i
s
o
l
at
e
d
i
n
p
r
e
v
io
u
s
r
e
s
ea
r
c
h
[
6
]
a
n
d
A
s
p
e
r
g
il
l
u
s
n
i
g
e
r
F
o
o
d
a
n
d
Nu
t
r
i
t
i
o
n
C
u
l
t
u
r
e
C
o
l
l
ec
t
i
o
n
(
FN
C
C
)
6
0
1
8
.
T
h
i
s
s
t
u
d
y
e
x
a
m
in
e
d
t
h
e
i
n
f
l
u
e
n
c
e
o
f
p
H
a
n
d
f
e
r
m
e
n
t
a
t
i
o
n
t
e
m
p
e
r
at
u
r
e
i
n
t
h
e
b
i
o
e
t
h
a
n
o
l
p
r
o
d
u
c
t
i
o
n
p
r
o
c
e
s
s
f
r
o
m
wi
l
d
c
as
s
av
a
u
s
i
n
g
t
h
e
SS
C
F
t
e
c
h
n
i
q
u
e
.
T
h
e
p
u
r
p
o
s
e
o
f
t
h
is
r
es
e
a
r
c
h
is
t
o
:
i
)
d
et
e
r
m
i
n
e
th
e
e
f
f
e
c
t
o
f
p
H
a
n
d
f
e
r
m
e
n
t
a
ti
o
n
t
e
m
p
e
r
a
t
u
r
e
o
n
t
h
e
c
h
a
r
a
c
t
e
r
is
t
i
cs
o
f
b
i
o
e
t
h
a
n
o
l
f
r
o
m
W
C
F
u
s
i
n
g
t
h
e
S
SC
F
t
e
c
h
n
i
q
u
e
;
a
n
d
i
i
)
d
e
t
e
r
m
i
n
e
t
h
e
a
p
p
r
o
p
r
i
a
t
e
p
H
a
n
d
f
e
r
m
e
n
t
a
t
i
o
n
t
e
m
p
e
r
at
u
r
e
t
o
p
r
o
d
u
c
e
b
i
o
e
t
h
a
n
o
l
f
r
o
m
W
C
F
u
s
i
n
g
a
S
SC
F
te
c
h
n
i
q
u
e
w
i
t
h
t
h
e
b
e
s
t
c
h
a
r
a
ct
e
r
is
t
i
cs
.
2.
RE
S
E
ARCH
M
E
T
H
O
D
2
.
1
.
Resea
rc
h
des
ig
n
T
h
e
ex
p
er
im
e
n
tal
d
esig
n
in
th
is
s
tu
d
y
u
s
ed
a
f
ac
to
r
ial
r
an
d
o
m
ized
b
lo
c
k
d
esig
n
(
R
B
D)
with
two
f
ac
to
r
s
.
T
h
e
f
ir
s
t
f
ac
to
r
was
th
e
in
itial
p
H
(
P)
,
wh
ich
co
n
s
is
ts
o
f
th
r
ee
lev
els,
n
am
ely
4
.
5
,
5
.
0
,
an
d
5
.
5
.
T
h
e
s
ec
o
n
d
f
ac
t
o
r
was
f
er
m
en
tatio
n
tem
p
er
atu
r
e
(
T
)
,
wh
ich
co
n
s
is
t
s
o
f
th
r
e
e
lev
els,
n
am
ely
3
0
,
3
2
.
5
,
an
d
3
5
°C
.
B
ased
o
n
th
e
co
m
b
in
atio
n
o
f
th
e
two
f
ac
to
r
s
ab
o
v
e,
n
in
e
tr
ea
tm
en
ts
wer
e
o
b
tain
ed
,
an
d
ea
ch
tr
ea
tm
en
t
was
g
r
o
u
p
ed
in
to
two
g
r
o
u
p
s
b
ased
o
n
p
r
o
ce
s
s
in
g
tim
e,
s
o
th
at
eig
h
teen
tr
ea
tm
en
t
u
n
its
wer
e
o
b
tain
ed
.
T
h
e
m
ain
in
g
r
ed
ien
t
u
s
ed
was
wild
ca
s
s
av
a
,
wh
ich
was
o
b
tain
ed
f
r
o
m
Kar
an
g
asem
Dis
tr
ict,
Kar
an
g
asem
R
eg
en
cy
,
B
ali
,
I
n
d
o
n
esia
.
T
h
e
wild
ca
s
s
av
a
o
b
tain
ed
was
p
r
o
ce
s
s
ed
in
to
W
C
F
th
r
o
u
g
h
a
p
r
o
ce
s
s
o
f
cu
ttin
g
,
s
o
ak
in
g
,
d
r
y
in
g
,
g
r
i
n
d
in
g
,
an
d
s
iev
in
g
.
T
h
e
f
o
llo
win
g
ad
d
itio
n
al
m
ater
ials
wer
e
u
tili
ze
d
in
th
is
s
tu
d
y
:
A
s
p
erg
illu
s
n
ig
er
FNC
C
6
0
1
8
,
alo
n
g
with
lo
ca
l
m
ic
r
o
b
es
C
a
n
d
id
a
o
r
th
o
p
s
ilo
s
is
R
5
I
3
an
d
K
leb
s
iella
va
r
iico
la
R
5
I
4
,
wh
ich
wer
e
o
b
tain
ed
f
r
o
m
t
h
e
B
io
in
d
u
s
tr
y
L
ab
o
r
at
o
r
y
at
th
e
Dep
ar
tm
e
n
t
o
f
Ag
r
o
i
n
d
u
s
tr
ial
T
ec
h
n
o
lo
g
y
,
Facu
lty
o
f
Ag
r
ic
u
ltu
r
al
T
ec
h
n
o
lo
g
y
,
U
d
ay
an
a
U
n
iv
er
s
ity
.
Evaluation Warning : The document was created with Spire.PDF for Python.
I
n
t J Ad
v
Ap
p
l Sci
I
SS
N:
2252
-
8
8
1
4
Temp
era
tu
r
e
a
n
d
p
H
effec
ts
o
n
b
io
eth
a
n
o
l
p
r
o
d
u
ctio
n
fr
o
m
w
ild
ca
s
s
a
va
…
(
I
d
a
A
y
u
P
r
id
a
r
i Ta
n
tr
i
)
229
2
.
2
.
Resea
rc
h
pro
ce
du
re
2
.
2
.
1
.
P
re
pa
ra
t
i
o
n
m
a
t
er
i
a
ls
W
ild
ca
s
s
av
a
tu
b
er
s
wer
e
p
ee
led
,
cu
t
in
to
s
m
aller
p
i
ec
es,
wash
ed
th
o
r
o
u
g
h
ly
,
s
o
ak
ed
a
n
d
co
n
tin
u
o
u
s
ly
s
tirr
ed
f
o
r
ap
p
r
o
x
im
ately
7
2
h
with
wate
r
was r
ep
lace
d
p
er
io
d
ically
ev
er
y
2
4
h
.
T
h
e
tu
b
er
s
wer
e
th
en
s
u
n
-
d
r
ied
,
th
en
d
r
ie
d
u
s
i
n
g
an
o
v
en
at
8
5
°C
f
o
r
5
-
7
h
.
T
h
e
d
r
ied
wild
ca
s
s
av
a
p
iece
s
wer
e
th
en
g
r
o
u
n
d
u
s
in
g
a
b
len
d
er
an
d
s
iev
ed
wit
h
a
6
0
-
m
esh
s
iev
e.
2
.
2
.
2
.
M
icro
o
rg
a
nis
m
s
a
nd
cult
ure
m
edium
C
a
n
d
id
a
o
r
th
o
p
s
ilo
s
is
R
5
I
3
an
d
K
leb
s
iella
va
r
iico
la
R
5
I
4
wer
e
r
eju
v
e
n
at
ed
u
s
in
g
y
e
ast
ex
tr
ac
t
p
ep
to
n
e
g
l
u
co
s
e
(
YPG)
m
ed
iu
m
with
a
co
m
p
o
s
itio
n
o
f
0
.
4
5
%
y
ea
s
t
ex
tr
ac
t,
0
.
7
5
%
p
ep
to
n
e
,
an
d
5
%
g
lu
c
o
s
e
in
d
is
till
ed
wate
r
[
3
]
,
[
6
]
,
[
1
3
]
.
T
h
r
ee
m
illi
liter
s
o
f
ea
ch
is
o
late
cu
ltu
r
e
wer
e
in
o
cu
lated
i
n
to
5
0
m
L
th
at
h
ad
b
ee
n
s
ter
ilized
an
d
in
cu
b
ated
at
3
5
°C
f
o
r
2
4
h
.
Pro
p
ag
atio
n
is
o
lates
ar
e
r
e
-
in
o
cu
lated
in
t
o
2
5
0
m
L
o
f
liq
u
id
YPG
m
ed
iu
m
f
o
r
2
4
h
at
r
o
o
m
tem
p
er
atu
r
e
an
d
1
2
5
r
p
m
a
g
itatio
n
f
o
r
ce
ll
g
r
o
wth
.
Af
ter
th
e
g
r
o
win
g
c
u
ltu
r
e
is
o
b
tain
ed
,
it is
ce
n
tr
if
u
g
ed
,
a
n
d
th
e
p
ellet
is
co
llected
[
3
]
,
[
1
3
]
.
A
s
p
erg
illu
s
n
ig
er
FN
C
C
6
0
1
8
was
r
eju
v
en
ated
an
d
p
r
o
p
a
g
at
ed
u
s
in
g
p
o
tato
d
ex
t
r
o
s
e
b
r
o
th
(
PDB
)
m
ed
ia.
A
t
th
e
r
eju
v
en
atio
n
s
tag
e
,
th
e
cu
ltu
r
e
was
in
c
u
b
ated
f
o
r
4
8
-
7
2
h
at
3
5
°C
in
an
in
cu
b
at
o
r
.
T
h
en
tr
a
n
s
f
er
r
ed
to
n
ew
PDB
m
ed
ia
f
o
r
p
r
o
p
ag
atio
n
an
d
in
c
u
b
ated
f
o
r
9
6
h
,
at
1
2
5
r
p
m
,
at
r
o
o
m
tem
p
er
atu
r
e
±30
°C
,
it will p
r
o
d
u
ce
ac
tiv
e
ce
lls
o
f
A
s
p
erg
illu
s
n
ig
er
FNC
C
6
0
1
8
[
1
4
]
.
2
.
2
.
3
.
Sin
g
le
-
s
t
a
g
e
o
f
SS
CF
in o
ne
s
ing
le
bio
re
a
ct
o
r
SS
C
F
ca
r
r
ied
o
u
t
r
ef
er
s
to
th
e
p
r
o
ce
d
u
r
e
u
s
ed
b
y
B
er
m
ejo
et
a
l.
[
1
5
]
an
d
Mo
s
h
i
et
a
l.
[
1
6
]
.
T
h
e
in
itial
s
tag
e
b
eg
in
s
with
weig
h
in
g
1
0
0
g
o
f
W
C
F
an
d
ad
d
in
g
7
0
0
m
L
o
f
d
is
till
ed
wate
r
,
o
r
u
s
in
g
a
r
atio
o
f
1
:7
f
o
r
o
n
e
f
er
m
en
tatio
n
m
ed
i
um
.
Gela
tin
izatio
n
at
8
0
°C
to
f
o
r
m
a
th
ick
s
lu
r
r
y
,
ad
ju
s
t
th
e
s
u
b
s
tr
ate
p
H
to
p
H
4
.
5
,
5
.
0
,
a
n
d
5
.
5
u
s
in
g
citr
ate
b
u
f
f
er
,
an
d
tr
an
s
f
er
2
0
0
m
L
o
f
s
u
b
s
tr
ate
in
to
a
2
5
0
m
L
f
er
m
en
to
r
.
Ster
ilizatio
n
u
s
in
g
an
au
t
o
clav
e
at
1
2
1
°C
f
o
r
1
5
m
in
with
a
p
r
ess
u
r
e
o
f
1
atm
,
af
ter
wh
ich
it
w
as
co
o
led
to
r
o
o
m
tem
p
er
atu
r
e.
A
s
p
erg
illu
s
n
ig
e
r
FNC
C
6
0
1
8
an
d
K
leb
s
iella
va
r
iico
la
R
5
I
4
wer
e
ad
d
e
d
s
im
u
ltan
eo
u
s
ly
b
e
f
o
r
e
th
e
f
er
m
e
n
tatio
n
p
r
o
ce
s
s
o
n
d
ay
0
an
d
af
ter
f
o
u
r
d
ay
s
o
f
th
e
f
er
m
e
n
tatio
n
p
r
o
ce
s
s
C
a
n
d
i
d
a
o
r
th
o
p
s
ilo
s
is
R
5
I
3
was
ad
d
ed
to
th
e
s
u
b
s
tr
ate
wit
h
a
tr
ea
tm
e
n
t
co
n
ce
n
tr
atio
n
o
f
2
%
in
ea
c
h
ex
p
er
im
en
tal
u
n
it
.
T
h
e
f
er
m
en
tatio
n
was
p
er
f
o
r
m
ed
in
a
wate
r
b
atc
h
s
h
ak
er
at
3
0
,
3
2
.
5
,
an
d
3
5
°C
with
a
s
tirr
in
g
s
p
ee
d
o
f
1
0
0
r
p
m
f
o
r
8
h
.
T
h
e
f
er
m
en
tatio
n
was th
en
f
ilter
ed
an
d
an
aly
ze
d
.
T
h
e
d
is
till
atio
n
p
r
o
ce
s
s
in
th
i
s
r
esear
ch
r
ef
er
s
t
o
th
e
r
esear
ch
p
r
o
ce
d
u
r
e
o
f
Gu
n
a
m
et
a
l
.
[
6
]
with
m
o
d
if
icatio
n
s
.
A
f
ter
th
e
f
er
m
en
tatio
n
p
r
o
ce
s
s
,
a
f
iltra
tio
n
p
r
o
ce
s
s
was
p
er
f
o
r
m
ed
u
s
in
g
a
d
is
till
ato
r
to
s
ep
ar
ate
th
e
f
e
r
m
en
tatio
n
f
iltr
ate
an
d
t
h
e
s
o
lid
s
.
T
h
e
f
iltra
t
e
was
m
ea
s
u
r
ed
with
a
g
r
ad
u
ated
cy
lin
d
e
r
,
a
n
d
th
en
a
m
u
ltis
tag
e
d
is
till
atio
n
was
p
er
f
o
r
m
ed
at
a
tem
p
er
atu
r
e
o
f
1
0
0
°C
f
o
r
th
e
f
ir
s
t
b
o
ttle
an
d
8
0
°C
f
o
r
th
e
s
ec
o
n
d
b
o
ttle.
Af
ter
d
is
till
atio
n
,
th
e
eth
a
n
o
l c
o
n
ten
t w
as a
n
a
ly
ze
d
.
2
.
3
.
Ana
ly
t
ica
l m
e
t
ho
ds
T
h
e
v
ar
iab
les
o
b
s
er
v
ed
in
th
is
r
esear
ch
in
clu
d
e
,
in
th
e
in
itial
s
tag
e
b
ef
o
r
e
f
er
m
en
tatio
n
,
p
H
an
aly
s
is
was
ca
r
r
ied
o
u
t
u
s
in
g
a
p
H
m
eter
to
m
ea
s
u
r
e
th
e
i
n
itial
p
H
b
ef
o
r
e
f
er
m
e
n
tatio
n
[
1
7
]
,
an
d
to
tal
s
o
lu
b
le
s
o
lid
s
an
aly
s
is
was
ca
r
r
ied
o
u
t
u
s
in
g
a
h
an
d
r
ef
r
ac
t
o
m
eter
to
m
ea
s
u
r
e
th
e
in
itial
to
tal
s
o
lu
b
le
s
o
lid
s
[
1
8
]
.
Af
ter
th
e
f
er
m
en
tatio
n
p
r
o
ce
s
s
,
p
H
an
a
ly
s
is
,
to
tal
s
o
lu
b
le
s
o
lid
s
an
aly
s
is
,
an
d
r
ed
u
cin
g
s
u
g
a
r
an
al
y
s
is
ar
e
p
er
f
o
r
m
ed
u
s
in
g
a
u
ltra
v
io
let
-
v
is
ib
le
(
UV
-
Vis
)
s
p
ec
tr
o
p
h
o
to
m
eter
to
m
ea
s
u
r
e
th
e
r
esu
lts
o
f
r
ed
u
cin
g
s
u
g
ar
p
r
o
d
u
ce
d
d
u
r
in
g
th
e
f
er
m
e
n
tatio
n
p
r
o
ce
s
s
[
6
]
,
[
1
3
]
.
T
h
e
n
,
a
f
ter
t
h
e
d
is
till
at
io
n
p
r
o
ce
s
s
,
eth
an
o
l
co
n
ten
t
an
aly
s
is
was
p
er
f
o
r
m
ed
u
s
in
g
an
alco
h
o
l
m
eter
to
d
eter
m
in
e
th
e
eth
an
o
l
p
r
o
d
u
ce
d
[
1
3
]
.
I
n
g
e
n
er
al,
th
e
f
lo
w
d
iag
r
am
o
f
th
e
b
io
eth
an
o
l
p
r
o
d
u
ctio
n
e
x
p
er
i
m
en
t is p
r
esen
ted
in
Fig
u
r
e
1
(
s
ee
Ap
p
en
d
ix
)
.
2
.
4
.
Da
t
a
a
na
ly
s
is
Data
wer
e
an
aly
ze
d
u
s
in
g
th
e
p
ar
am
etr
ic
an
aly
s
is
o
f
v
ar
ian
ce
(
ANOV
A)
te
s
t
at
a
s
ig
n
if
ic
an
ce
lev
el
o
f
p
<
0
.
0
5
.
W
h
en
s
ig
n
i
f
ican
t
d
if
f
er
en
ce
s
wer
e
o
b
s
er
v
ed
a
m
o
n
g
tr
ea
tm
en
ts
,
f
u
r
th
er
an
al
y
s
is
was
co
n
d
u
cted
u
s
in
g
Du
n
ca
n
’
s
m
u
ltip
le
r
an
g
e
test
(
DM
R
T
)
.
All
s
tati
s
ti
ca
l
p
r
o
ce
s
s
in
g
was
p
er
f
o
r
m
ed
with
th
e
Min
itab
s
o
f
twar
e
p
r
o
g
r
am
.
3.
RE
SU
L
T
S AN
D
D
I
SCU
SS
I
O
N
3
.
1
.
pH
T
h
e
r
esu
lts
o
f
v
ar
ian
ce
an
aly
s
is
s
h
o
wed
th
at
p
H,
f
er
m
en
ta
tio
n
tem
p
er
atu
r
e,
a
n
d
th
eir
in
ter
ac
tio
n
s
h
ad
a
v
e
r
y
s
ig
n
if
ica
n
t
ef
f
ec
t
(
p
<0
.
0
1
)
o
n
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e
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1
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t
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r
e
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5
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r
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Evaluation Warning : The document was created with Spire.PDF for Python.
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d
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5
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5
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2
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5
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an
d
3
5
°C
.
Fro
m
th
ese
r
esu
lts
,
th
e
p
H
in
ea
ch
tr
ea
tm
en
t
in
cr
ea
s
ed
d
u
r
in
g
th
e
f
er
m
en
tatio
n
p
r
o
ce
s
s
d
u
e
to
th
e
b
y
-
p
r
o
d
u
cts
o
f
th
e
f
er
m
en
tatio
n
p
r
o
ce
s
s
in
th
e
f
o
r
m
o
f
o
r
g
an
ic
ac
id
s
.
T
h
is
is
s
u
p
p
o
r
ted
b
y
th
e
s
tatem
en
t
o
f
Ma
et
a
l.
[
1
9
]
th
a
t
th
e
f
er
m
e
n
tatio
n
p
r
o
ce
s
s
p
r
o
d
u
ce
s
b
y
-
p
r
o
d
u
cts
in
t
h
e
f
o
r
m
o
f
o
r
g
a
n
ic
ac
id
s
,
o
lig
o
s
ac
ch
ar
id
es,
am
y
lase,
lip
ase,
ce
llu
lase,
x
y
lase
,
an
d
p
r
o
tease.
T
h
is
s
tatem
en
t
is
r
e
in
f
o
r
ce
d
b
y
th
e
v
iew
o
f
Gh
o
r
i
et
a
l.
[
2
0
]
,
wh
ich
s
tates
th
at
th
e
f
er
m
en
tatio
n
p
r
o
ce
s
s
n
o
t
o
n
ly
p
r
o
d
u
ce
s
eth
an
o
l
b
u
t
also
o
r
g
an
ic
ac
id
c
o
m
p
o
u
n
d
s
,
th
er
e
b
y
in
cr
ea
s
in
g
th
e
ac
id
ity
o
f
th
e
s
u
b
s
tr
ate.
T
ab
le
1
.
Av
e
r
ag
e
p
H
v
alu
e
o
f
f
er
m
en
tatio
n
r
esu
lts
pH
Te
mp
e
r
a
t
u
r
e
(
°C
)
30
°C
3
2
.
5
°C
35
°C
4
.
5
3
.
3
8
±
0
.
1
1
cd
3
.
5
5
±
0
.
0
7
b
3
.
7
3
±
0
.
1
1
a
5
.
0
3
.
2
8
±
0
.
0
4
de
3
.
3
8
±
0
.
1
1
cd
3
.
4
5
±
0
.
0
7
bc
5
.
5
3
.
1
8
±
0
.
0
4
e
3
.
2
3
±
0
.
0
4
de
3
.
3
0
±
0
.
0
0
c
de
N
o
t
e
:
m
e
a
n
s
f
o
l
l
o
w
e
d
b
y
d
i
f
f
e
r
e
n
t
l
e
t
t
e
r
s
w
i
t
h
i
n
t
h
e
s
a
m
e
r
o
w
o
r
c
o
l
u
m
n
a
r
e
s
i
g
n
i
f
i
c
a
n
t
l
y
d
i
f
f
e
r
e
n
t
(
p
<
0
.
0
5
)
I
n
g
en
er
al,
p
H
is
an
im
p
o
r
tan
t
f
ac
to
r
in
th
e
f
er
m
en
tatio
n
p
r
o
ce
s
s
b
ec
au
s
e
it
in
f
lu
en
ce
s
th
e
ac
tiv
ity
o
f
m
icr
o
o
r
g
an
is
m
s
[
2
1
]
,
an
d
p
r
o
p
er
p
H
is
th
e
k
ey
to
m
icr
o
b
ial
g
r
o
wth
an
d
m
eta
b
o
lis
m
[
2
2
]
.
Acc
o
r
d
in
g
to
Gu
n
am
et
a
l.
[
6
]
,
th
e
o
p
tim
al
p
H
v
alu
e
f
o
r
R
5
I
3
a
n
d
R
5
I
4
ac
tiv
ity
is
p
H
5
.
Oth
er
r
esea
r
ch
s
tates
th
at
th
e
o
p
tim
al
p
H
f
o
r
th
e
ac
tiv
e
en
z
y
m
e
-
p
r
o
d
u
ci
n
g
en
zy
m
es
f
r
o
m
A
s
p
erg
illu
s
n
ig
er
is
p
H
5
-
5
.
5
[
2
3
]
.
R
esear
ch
b
y
T
u
r
h
an
et
a
l.
[
2
4
]
s
tated
th
at
p
H
-
co
n
tr
o
lled
tr
ea
tm
en
ts
wer
e
ab
le
to
p
r
o
d
u
ce
h
ig
h
er
eth
an
o
l
co
n
ce
n
tr
atio
n
s
th
an
tr
ea
tm
e
n
ts
with
o
u
t p
H
c
o
n
tr
o
l.
T
h
e
i
n
itial
p
H
v
alu
e
o
f
t
h
is
s
tu
d
y
was
s
et
at
p
H
v
ar
iati
o
n
s
o
f
4
.
5
,
5
.
0
,
an
d
5
.
5
in
o
r
d
er
to
cr
ea
te
a
n
o
p
tim
al
f
er
m
en
tatio
n
m
ed
iu
m
f
o
r
t
h
e
m
icr
o
o
r
g
an
is
m
s
.
3
.
2
.
T
o
t
a
l
s
o
lub
le
s
o
lid
s
T
h
e
r
esu
lts
o
f
th
e
ANOVA
s
h
o
wed
th
at
th
e
p
H
an
d
f
er
m
en
t
atio
n
tem
p
er
atu
r
e
tr
ea
tm
en
ts
h
ad
a
r
ea
l
ef
f
ec
t
(
p
>
0
.
0
5
)
,
wh
ile
th
e
p
H
an
d
in
ter
ac
tio
n
h
a
d
n
o
s
ig
n
if
i
ca
n
t
ef
f
ec
t
(
p
>0
.
0
5
)
o
n
t
h
e
to
t
al
v
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e
o
f
s
o
lu
b
le
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o
lid
s
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d
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ce
d
d
u
r
in
g
th
e
S
SC
F
p
r
o
ce
s
s
.
T
h
e
av
er
ag
e
v
alu
e
o
f
to
tal
s
o
lu
b
le
s
o
lid
s
i
n
th
e
f
er
m
en
tatio
n
p
r
o
ce
s
s
ca
n
b
e
s
ee
n
in
T
ab
le
2
.
T
h
e
r
esu
lts
in
T
a
b
le
2
s
h
o
w
th
e
av
er
ag
e
to
tal
s
o
lu
b
le
s
o
li
d
s
o
cc
u
r
r
in
g
at
ea
ch
p
H
an
d
f
er
m
en
tatio
n
tem
p
e
r
at
u
r
e.
T
h
e
a
v
er
ag
e
v
alu
e
o
f
to
ta
l
s
o
lu
b
le
s
o
lid
s
is
9
.
3
–
7
.
9
°B
r
ix
.
T
h
e
tr
ea
tm
en
t o
f
p
H
5
.
5
was
n
o
t
s
ig
n
if
ica
n
tly
d
if
f
e
r
en
t
f
r
o
m
p
H
4
.
5
an
d
5
.
0
.
T
h
e
h
ig
h
est
tem
p
er
atu
r
e
tr
ea
tm
en
t
was
at
3
2
.
5
°C
,
b
u
t
n
o
t
s
ig
n
if
ican
tly
d
if
f
e
r
en
t
f
r
o
m
3
5
°C
,
a
n
d
lo
west
was
at
3
5
°C
.
I
n
th
e
f
er
m
en
tatio
n
p
r
o
ce
s
s
,
s
u
g
ar
is
m
etab
o
lized
b
y
R
5
I
3
in
to
alco
h
o
l
an
d
C
O
2
,
ca
u
s
in
g
lo
w
to
tal
s
o
lu
b
le
s
o
l
id
s
.
As
s
tated
b
y
Su
jan
a
et
a
l.
[
1
3
]
,
th
e
lo
w
to
tal
s
o
lu
b
le
s
o
lid
s
ar
e
th
o
u
g
h
t
to
b
e
b
ec
au
s
e
d
u
r
in
g
t
h
e
f
er
m
en
tatio
n
p
r
o
ce
s
s
,
s
u
g
ar
,
wh
ic
h
is
th
e
d
o
m
in
an
t
s
o
lid
co
m
p
o
n
en
t
in
t
h
e
m
e
d
iu
m
,
is
m
etab
o
lized
b
y
y
ea
s
t
in
t
o
alco
h
o
l
a
n
d
C
O
2
,
th
en
u
s
ed
as
a
ca
r
b
o
n
s
o
u
r
ce
b
y
latic
ac
id
b
ac
ter
ia
(
L
AB
)
,
s
o
th
at
th
e
to
tal
s
o
lid
s
co
n
ten
t
d
is
s
o
lv
ed
to
lo
w.
T
h
is
is
r
ein
f
o
r
ce
d
b
y
th
e
s
tatem
en
t
o
f
Gu
n
am
et
a
l.
[
6
]
,
wh
er
e
th
e
d
ec
r
ea
s
e
in
to
tal
s
o
lu
b
le
s
o
lid
s
d
u
r
in
g
s
to
r
ag
e
is
d
u
e
to
th
e
co
n
v
er
s
io
n
o
f
th
e
s
u
g
ar
co
n
tain
ed
in
to
a
lco
h
o
l,
al
d
eh
y
d
es,
an
d
am
i
n
o
ac
id
s
.
T
h
is
o
p
in
io
n
is
s
u
p
p
o
r
ted
b
y
Gu
n
a
m
et
a
l
.
[
2
5
]
s
tated
th
at
th
e
l
o
wer
th
e
to
tal
s
o
lid
s
co
n
ten
t,
th
e
h
ig
h
er
t
h
e
eth
an
o
l c
o
n
ten
t.
T
h
e
d
ec
r
ea
s
e
in
to
tal
s
o
lu
b
le
s
o
lid
s
also
o
cc
u
r
r
ed
d
u
e
to
th
e
b
r
ea
k
d
o
wn
o
f
g
lu
c
o
s
e
b
y
m
icr
o
b
es
in
to
alco
h
o
l.
T
h
e
a
d
d
ed
m
icr
o
o
r
g
a
n
is
m
s
h
av
e
th
e
ab
ilit
y
to
p
r
o
d
u
ce
α
-
am
y
lase
an
d
g
lu
co
am
y
l
ase,
wh
ich
ar
e
u
s
ed
to
co
n
v
er
t
t
h
e
am
y
lo
s
e
an
d
a
m
y
lo
p
ec
tin
co
n
tain
e
d
in
s
tar
c
h
in
to
s
u
g
ar
s
(
g
alac
to
s
e,
f
r
u
c
to
s
e,
an
d
g
lu
c
o
s
e)
d
u
r
in
g
ae
r
o
b
ic
o
r
a
n
ae
r
o
b
ic
f
er
m
en
tatio
n
,
co
n
v
er
ted
i
n
to
eth
an
o
l,
C
O
2
,
an
d
o
th
e
r
b
y
-
p
r
o
d
u
cts
[
2
6
]
.
T
h
e
f
u
n
g
al
m
icr
o
o
r
g
a
n
is
m
A
s
p
erg
illu
s
n
ig
er
is
ca
p
ab
le
o
f
p
r
o
d
u
cin
g
g
lu
c
o
am
y
lase,
c
ellu
lase,
x
y
lan
ase,
am
y
lase,
p
r
o
tease,
an
d
lip
ase
[
2
7
]
,
[
2
8
]
.
T
h
is
ab
ilit
y
is
r
eq
u
ir
ed
in
th
e
f
er
m
e
n
tatio
n
p
r
o
ce
s
s
to
co
n
v
er
t
s
tar
ch
to
eth
an
o
l.
T
ab
le
2
.
Av
e
r
ag
e
v
al
u
e
o
f
t
o
tal
s
o
lu
b
le
s
o
lid
s
f
r
o
m
f
er
m
en
tat
io
n
(
°B
r
i
x
)
pH
Te
mp
e
r
a
t
u
r
e
(
°C
)
A
v
e
r
a
g
e
30
°C
3
2
.
5
°C
35
°C
4
.
5
7
.
9
0
±
0
.
4
2
9
.
3
0
±
0
.
5
7
8
.
0
5
±
0
.
3
5
8
.
4
2
±
0
.
1
1
a
5
.
0
7
.
9
5
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Evaluation Warning : The document was created with Spire.PDF for Python.
I
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v
Ap
p
l Sci
I
SS
N:
2252
-
8
8
1
4
Temp
era
tu
r
e
a
n
d
p
H
effec
ts
o
n
b
io
eth
a
n
o
l
p
r
o
d
u
ctio
n
fr
o
m
w
ild
ca
s
s
a
va
…
(
I
d
a
A
y
u
P
r
id
a
r
i Ta
n
tr
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)
231
3.
3
.
Reducing
s
ug
a
r
T
h
e
r
esu
lts
o
f
v
ar
ian
ce
an
aly
s
is
s
h
o
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th
at
p
H,
f
er
m
en
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tio
n
tem
p
e
r
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r
e,
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d
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ci
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en
tatio
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ca
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T
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3
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r
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lts
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T
ab
le
3
s
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o
w
th
e
av
er
a
g
e
v
alu
e
o
f
r
e
d
u
cin
g
s
u
g
ar
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o
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ea
c
h
p
H
an
d
f
er
m
en
tatio
n
tem
p
er
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e
tr
e
atm
en
t.
T
h
e
h
ig
h
est
av
er
ag
e
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o
f
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u
ci
n
g
s
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g
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f
r
o
m
f
er
m
e
n
t
atio
n
o
cc
u
r
r
ed
in
th
e
p
H
5
.
5
tr
ea
tm
en
t
with
a
f
er
m
en
tatio
n
tem
p
er
atu
r
e
o
f
3
2
.
5
°C
,
p
r
o
d
u
cin
g
3
.
6
6
±
0
.
1
2
m
g
/m
L
.
T
h
e
lo
west
av
er
ag
e
v
alu
e
o
f
r
ed
u
cin
g
s
u
g
ar
r
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f
r
o
m
f
er
m
en
tat
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n
o
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r
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in
th
e
p
H
4
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5
t
r
e
atm
en
t
with
a
f
er
m
en
tatio
n
te
m
p
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o
f
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0
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p
r
o
d
u
cin
g
0
.
9
4
±
0
.
0
3
m
g
/m
L
.
W
ith
in
cr
ea
s
in
g
p
H
an
d
f
e
r
m
en
tatio
n
tem
p
er
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r
e,
t
h
e
v
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wh
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at
th
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p
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T
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e
r
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m
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d
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s
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ely
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tr
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o
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an
o
l it
ca
n
p
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d
u
ce
[
2
9
]
.
T
ab
le
3
.
Av
e
r
ag
e
v
al
u
e
o
f
r
ed
u
cin
g
s
u
g
ar
f
r
o
m
f
er
m
en
tatio
n
pH
Te
mp
e
r
a
t
u
r
e
(
°C
)
30
°C
3
2
.
5
°C
35
°C
4
.
5
0
.
9
4
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0
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0
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3
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0
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1
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1
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6
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2
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2
0
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3
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1
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6
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o
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t
l
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h
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me
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t
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p
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T
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m
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A
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K
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s
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r
tan
t
r
o
le
in
p
r
o
d
u
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g
α
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y
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m
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ch
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t
o
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l
o
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e
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d
am
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lo
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ts
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4
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ly
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ts
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1
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6
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l
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s
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h
e
α
-
1
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4
g
ly
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d
s
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s
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ch
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y
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s
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r
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ct
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r
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to
p
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ce
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[
3
0
]
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ze
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α
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1
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4
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ly
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ly
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to
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p
r
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s
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if
icatio
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[
3
0
]
.
Fer
m
en
t
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g
lu
co
s
e
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tili
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d
b
y
m
icr
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m
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s
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r
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f
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t
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d
p
r
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ce
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an
o
l b
y
-
p
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d
u
cts an
d
o
r
g
an
ic
ac
id
s
[
6
]
,
[
1
3
]
.
3.
4
.
E
t
ha
no
l
co
nte
nt
T
h
e
r
esu
lts
o
f
th
e
ANOVA
s
h
o
wed
th
at
th
e
p
H
a
n
d
f
e
r
m
en
t
atio
n
tem
p
er
atu
r
e
h
ad
a
v
er
y
s
ig
n
if
ican
t
ef
f
ec
t
(
p
<0
.
0
1
)
,
wh
ile
th
e
in
t
er
ac
tio
n
h
ad
a
s
ig
n
if
ica
n
t
ef
f
ec
t
(
p
<0
.
0
5
)
o
n
t
h
e
v
alu
e
o
f
th
e
eth
an
o
l
co
n
ten
t
p
r
o
d
u
ce
d
d
u
r
in
g
th
e
SS
C
F
p
r
o
ce
s
s
.
T
h
e
av
er
ag
e
v
alu
e
o
f
e
th
an
o
l
co
n
ten
t
in
t
h
e
f
er
m
en
ta
tio
n
p
r
o
ce
s
s
ca
n
b
e
s
ee
n
in
T
ab
le
4
.
T
h
e
r
esu
lts
in
T
ab
le
4
s
h
o
w
th
e
av
er
a
g
e
v
alu
e
o
f
eth
an
o
l
c
o
n
ten
t
o
cc
u
r
r
in
g
at
ea
ch
p
H
an
d
f
er
m
en
tatio
n
tem
p
er
atu
r
e.
T
h
e
h
ig
h
est
a
v
er
ag
e
eth
an
o
l
v
al
u
e
was
at
p
H
4
.
5
with
a
f
er
m
en
tatio
n
tem
p
e
r
atu
r
e
o
f
3
2
.
5
°C
,
p
r
o
d
u
cin
g
3
.
4
8
±
0
.
3
7
%
w/v
,
n
o
t
s
ig
n
if
ica
n
tly
d
if
f
er
en
t
f
r
o
m
p
H
4
.
5
at
3
0
°C
an
d
p
H
5
.
0
at
3
2
.
5
°C
.
T
h
e
lo
west
av
er
ag
e
eth
an
o
l
v
alu
e
was
at
p
H
5
.
5
with
a
f
er
m
en
tatio
n
tem
p
e
r
atu
r
e
o
f
3
2
.
5
°C
,
p
r
o
d
u
cin
g
1
.
3
5
±
0
.
2
4
%
w/v
,
b
u
t
was
n
o
t
s
ig
n
if
ican
tly
d
if
f
er
en
t
f
r
o
m
p
H
5
.
0
at
3
0
°C
an
d
3
5
°C
.
T
h
e
eth
an
o
l
co
n
ten
t
v
alu
e
ten
d
s
to
d
ec
r
ea
s
e
as
th
e
p
H
v
alu
e
i
n
cr
ea
s
es.
T
h
is
s
h
o
ws
th
at
at
p
H
5
.
5
,
th
e
g
r
o
wth
an
d
ac
tiv
ity
o
f
R
5
I
3
ar
e
n
o
t
o
p
tim
al.
R
5
I
3
en
ter
s
th
e
s
tatio
n
ar
y
p
h
ase
o
r
b
ec
o
m
es
less
ef
f
icien
t
at
p
H
5
.
5
.
T
h
is
p
h
ase
is
ca
u
s
ed
b
y
th
e
lev
el
o
f
g
lu
co
s
e
u
s
ed
as
a
ca
r
b
o
n
s
o
u
r
ce
f
o
r
y
ea
s
t
ce
lls
ten
d
in
g
to
d
ec
r
e
ase,
p
r
ev
en
tin
g
th
e
y
ea
s
t
f
r
o
m
b
r
ea
k
in
g
d
o
wn
in
t
o
eth
an
o
l
an
d
ca
u
s
in
g
s
o
m
e
o
f
th
e
y
ea
s
t
to
d
ie
[
3
1
]
.
I
f
th
e
p
H
is
in
cr
ea
s
ed
,
th
e
g
r
o
wth
r
ate
will
d
ec
r
ea
s
e
,
an
d
ev
e
n
tu
ally
,
g
r
o
wth
will
s
to
p
alto
g
eth
e
r
.
Gr
o
wth
a
r
r
est
c
an
b
e
ca
u
s
ed
b
y
a
r
ed
u
ctio
n
in
s
ev
e
r
al
ess
en
tial
n
u
tr
ien
ts
in
th
e
m
ed
iu
m
,
o
r
b
y
th
e
ac
cu
m
u
latio
n
o
f
au
to
t
o
x
in
s
in
th
e
m
ed
iu
m
,
o
r
a
co
m
b
in
atio
n
o
f
b
o
th
[
1
3
]
.
Me
an
wh
ile,
th
e
eth
an
o
l
co
n
ten
t
v
alu
e
ten
d
s
to
in
c
r
ea
s
e
with
in
cr
ea
s
in
g
f
er
m
en
tatio
n
tem
p
er
atu
r
e
.
T
ab
le
4
.
Av
e
r
ag
e
v
al
u
e
o
f
et
h
an
o
l
co
n
te
n
t (
%
w/v
)
pH
Te
mp
e
r
a
t
u
r
e
(
°C
)
30
°C
3
2
.
5
°C
35
°C
4
.
5
2
.
7
8
±
0
.
4
2
a
bc
3
.
4
8
±
0
.
3
7
a
2
.
3
7
±
0
.
5
0
cd
5
.
0
2
.
0
0
±
0
.
2
4
de
3
.
2
3
±
0
.
6
5
ab
2
.
1
1
±
0
.
2
6
c
de
5
.
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Evaluation Warning : The document was created with Spire.PDF for Python.
I
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2
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I
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v
Ap
p
l Sci
,
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1
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No
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1
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Ma
r
c
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232
T
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m
tem
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er
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r
e
f
o
r
g
r
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wth
an
d
ac
tiv
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f
A
s
p
erg
illu
s
n
ig
er
is
3
0
°C
.
T
h
e
o
p
tim
u
m
tem
p
er
atu
r
e
f
o
r
C
a
n
d
id
a
o
r
th
o
p
s
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iico
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wth
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wev
er
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in
th
is
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y
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it
was
s
lig
h
tly
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if
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tem
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e
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5
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,
th
e
m
icr
o
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e
s
co
u
ld
s
till
wo
r
k
,
an
d
th
is
was
th
e
b
est
r
esu
lt
f
o
r
p
r
o
d
u
cin
g
th
e
h
ig
h
est
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l.
T
em
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er
at
u
r
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p
lay
s
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im
p
o
r
tan
t
r
o
le
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ec
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s
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it
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n
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ir
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tly
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f
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ec
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th
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icr
o
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ial
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ir
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tly
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f
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t
th
e
am
o
u
n
t
o
f
eth
a
n
o
l
p
r
o
d
u
ce
d
[
3
2
]
.
Ap
a
r
t
f
r
o
m
th
at,
th
e
d
ec
r
ea
s
e
in
b
io
eth
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l
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atio
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lu
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ce
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g
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th
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y
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,
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h
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f
e
r
m
en
tatio
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ate.
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h
en
th
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f
er
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ast
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ile
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ates,
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m
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id
ized
eth
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o
l to
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m
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id
Hata
et
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[
3
3
]
.
4.
CO
NCLU
SI
O
N
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ec
en
t
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er
v
atio
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icate
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t
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m
en
tatio
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tem
p
er
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h
a
d
a
s
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n
if
ican
t
ef
f
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t
o
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th
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p
H
v
alu
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d
u
c
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e,
an
d
eth
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u
t
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ad
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ig
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u
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SS
C
F
tech
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iq
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p
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tr
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en
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f
4
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5
with
a
f
er
m
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m
p
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2
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5
°C
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ig
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t p
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g
/m
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ess
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iv
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th
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ee
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to
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p
l
o
y
d
i
v
er
s
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f
er
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en
tatio
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co
n
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itio
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to
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iev
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o
p
tim
al
o
u
tco
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o
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s
eq
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en
tly
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es
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id
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as a
n
alter
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ativ
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aw
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ater
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an
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p
r
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d
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ctio
n
.
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NO
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DG
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au
th
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wo
u
l
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to
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Un
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T
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lab
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esp
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.
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UNDING
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u
n
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an
a
Un
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s
ity
th
r
o
u
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t
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“Pen
elitian
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AUTHO
R
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h
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u
r
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s
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C
o
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r
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to
r
R
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les
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a
x
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y
(
C
R
ed
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ize
in
d
iv
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d
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al
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th
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tio
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r
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h
ip
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p
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tes,
an
d
f
ac
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llab
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atio
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.
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m
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Aut
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Prid
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ak
Ag
u
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d
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C
:
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ter
ests
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h
at
co
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ld
in
f
lu
e
n
ce
th
e
r
esear
ch
f
in
d
in
g
s
.
T
h
is
wo
r
k
was
co
n
d
u
cted
in
d
e
p
e
n
d
en
tly
to
m
ain
tain
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b
jectiv
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d
in
teg
r
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in
r
ep
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tin
g
r
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lts
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elate
d
to
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d
ca
s
s
av
a
(
Ma
n
ih
o
t g
l
a
z
io
vii
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u
ll.
Ar
g
.
)
p
r
o
ce
s
s
in
g
an
d
an
aly
s
is
.
Evaluation Warning : The document was created with Spire.PDF for Python.
I
n
t J Ad
v
Ap
p
l Sci
I
SS
N:
2252
-
8
8
1
4
Temp
era
tu
r
e
a
n
d
p
H
effec
ts
o
n
b
io
eth
a
n
o
l
p
r
o
d
u
ctio
n
fr
o
m
w
ild
ca
s
s
a
va
…
(
I
d
a
A
y
u
P
r
id
a
r
i Ta
n
tr
i
)
233
DATA AV
AI
L
AB
I
L
I
T
Y
All d
ata
g
en
er
ated
o
r
an
al
y
ze
d
d
u
r
in
g
th
is
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tu
d
y
ar
e
i
n
clu
d
e
d
in
th
is
p
u
b
lis
h
ed
a
r
ticle.
RE
F
E
R
E
NC
E
S
[
1
]
N
.
H
a
n
d
r
a
,
A
.
K
a
s
i
m,
G
.
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n
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r
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,
“
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.
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[
2
]
M
.
K
r
a
j
a
n
g
,
K
.
M
a
l
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i
r
u
a
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s
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c
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.
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