Theobromine Content in Cocoa Pod Husk (Theobroma cacao) Fermented by Aspergillus spp. in Different of Chop Sizes and Fermentation Times
Theobromine Content in Cocoa Pod Husk (Theobroma
(
cacao)
Fermented by Aspergillus spp. in Different of Chop Sizes and
Fermentation Times
F. F. Munier1 & H. Hartadi2
1
Graduate Student of Animal Science Institute, Gadjah Mada University, Fauna
Street No. 3, Bulaksumur, Yogyakarta, 55281
2
Animal Science Faculty, Gadjah Mada University, Fauna Street No. 3,
Bulaksumur, Yogyakarta, 55281
Abstract
Theobromine is one of alkaloid in cocoa pod husk (CPH). Fresh CPH has
high theobromine content reach 0.40% (400.000 ppm) which is fresh CPH portion
in ruminant ration more than 30% to be affected of health. This research aim was
to know the change of theobromine content of CPH with different chop sizes were
fermented in the different time of incubation by Aspergillus spp. Three species of
Aspergillus spp. were used in this research i.e. A. niger, A. oryzae and A. ficuum
BPT. There were 4 chop sizes i.e.; irrigular size (A1), 1x5 cm (A2), 3x5 cm (A3), and
5x5 cm (A4). The incubation times were; 0, 5, 7, and 9 days, with three replications.
The solely parameter was theobromine content which were prepared according to
AOAC (1990) and analyze using HPLC (Chen et al., 2008). The result of research
showed that CPH fermented using A. niger, the lowest of theobromine content for
chop size A2 and incubated 9 days was 38.088 ppm. CPH fermented using A. oryzae
had the lowest of theobromine content in A2 and 9 days only 36.215 ppm. CPH
fermented with A. ficuum had the lowest of theobromine content in A2 and 9 days
reach 42.914 ppm. It was concluded that the best of CPH fermented
ermented for reducing of
theobromine content for 1x5 cm (A2) chop size with 9 days fermentation, fermented
by A. oryzae.
Keywords: theobromine,, cocoa pod husk, Aspergillus spp., chop sizes, incubation
times
Introducton
Cocoa pod husk (CPH) utlzaton s expected to be capable of feed fulfllng the
rumnant anmal requrement yearly, so the anmal wll not be lackng of nutrents for
ts lfe and producton. Ths condton has reason because cocoa trees have frut n
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
175
a long year whch s CPH abundant avalable n harvest season. Utlzaton of fresh
CPH as feed supplement has lmtng factor whch one alkalod s theobromne. CPH
has hgh of theobromne content or 3,7-dmethylxanthne (Mahyuddn and Bakre,
1993; Aregheore, 1999).
). The hgh concentraton theobromne affected the health of
anmal (Goldng, 1985). The theobromne content of CPH about 0.17-0.20% (Wong
and Hassan, 1988),
8),
), whle n cocoa beans reached 1.5-3.0%,
.5-3.0%,
5-3.0%,
.0%,
%, and n dry cocoa beans
reached 1.8%
.8% (Wllson, 1999)..
Decreasng of theobromne content can be done wth fermentaton method usng mcrobe agent such as fung. Some researchers used A. niger as decomposer
of CPH n sold fermentaton process (Yamaoka-Yano and Mazzafera, 1999; Huq,
2006) or A.. oryzae (Lubs et al., 2002). They found that CPH theobromne content
were reduced due to the role of enzyme actvty that secreted by Aspergillus spp.
Based on those results, n ths research we used A. niger, A. oryzae and A. ficuum n sold fermentaton wth CPH as a substrate. The am of ths research was
to decrease theobromne content of CPH n Laboratory scale that subsequently the
fermented CPH would used for alternatve forage, and also we studed about storage
tme of fermented CPH as feed.
Materals and Methods
Ths experment was fermented CPH usng three of Aspergillus spp..e. A. niger, A. oryzae and A. ficuum, n four types of chop szes .e. rregular sze (A1) as
control, 1 x 5 cm (A2), 3 x 5 cm (A3), and 5 x 5 cm (A4) and four knds of ncubaton
tmes .e. 0, 5, 7 and 9 days. CPH was ar dred for 8 hours after choppng, whle
powder fung were used as much as 1% and then added urea 1% of CPH weght.
Urea dssolved wth sterle water n rato 1 : 10 parts. Fresh CPH used 1 kg every
treatment whch ths was spread n plastc box (the frst coatng). CPH sprayed wth
urea and noculated of Aspergillus spp., the next wth same procedure was done for
the second and thrd coatngs.
After completon of fermentaton process, CPH was removed and to be ardred for 6 hours. The weghng of CPH and to be contnued wth samples dryng
usng oven n temperature of 55 oC for 4 days. About 100 g of fermented CPH was
dred usng freeze dryer for 4 days. Dred CPH was ground wth the mortal whch
was surrounded by ce to mantan optmal temperature and theobromne content,
and then followed by analyss of theobromne content of CPH. Analyss of theobromne content was recommended by EFSA (2008) usng Hgh-Performance Lqud
Chromatography (HPLC).
Results and Dscusson
Theobromne as alkalod compound cause btter n cocoa beans and CPH. Al-
176
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
kalod s organc component from plant whch has contan of ntrogen, commonly as
alkals and t has bology actvty. The alkalod n plant has functon for attact of pest
(Urch, 1994). Fermentas treatment can decrease CPH theobromne content. CPH
non fermented has hgher of theobromne content (Tablee 1).
).
Table 1. Theobromne content of cocoa pod husk fermented by A. niger (ppm)
Treatment of chop szes
Incubaton
tmes (days)
A1
A2
A3
A4
0
5
7
9
Average
157.560
76.008
38.545
38.477
77.648
134.410
71.093
38.103
38.088
70.424
171.554
78.857
46.680
43.837
85.232
186.193
81.778
52.697
45.727
91.599
Average
162.429
76.934
44.006
37.507
Note: A1= rregular sze, A2= 1 x 5 cm, A3= 3 x 5 cm, A4= 5 x 5 cm.
The fndng results showed that fermentaton process affected the decreasng
of CPH theobromne content. In A. niger fermentaton treatment, there was an effect
on CPH theobromne content due to dfferences of chop szes and ncubaton tmes.
Reducng of CPH chop sze was parallel to reducng of theobromne content. The
A2 and A4 chop szes showed average of the lowest and the hghest of theobromne
content, respectvely. On the other sde, ncreasng of ncubaton tme was nversed
wth decreasng of average of theobromne content. The lowest average of theobromne content was acheved through fermentaton at 9 days of ncubaton (reduced
76.91%). From ths experment, the lowest of theobromne content (38.088 ppm)
was acheved n A2 chop sze when ncubated durng 9 days.
Fermentaton
ermentaton usng A. oryzae n treatment of chop szes and ncbaton tmes
showed effect to decreasng of CPH theobromne content that was presented n Table 2. Reducng of CPH chop sze had correlaton wth reducng of theobromne
content. The A2 chop sze showed average of the lowest theobromne content, whle
A4 was the hghest theobromne content. The longer ncubaton tme supported reducng theobromne content. The lowest average of theobromne content was occur
at 9 days fermentaton (reduced 23.81%). From ths experment, the lowest of theobromne content (36.215 ppm) was acheved n A2 chop sze when ncubated durng
9 days.
The fndng results showed that fermentaton CPH usng A. ficuum affected
the decreasng of theobromne content (Table
Table 3). Decreasng of CPH chop sze had
relaton wth decreasng of theobromne content. The A2 chop sze showed average
of the lowest for theobromne content. On the other sde, ncreasng of ncubaton
tme was nversed wth decreasng of average of theobromne content. The lowest
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
177
Table 2. CPH theobromne content KBK nonfermented and fermented wth A. niger (ppm)
Chop szes
Incubaton
tmes (days)
A1
A2
A3
A4
0
5
7
9
Average
181.891
62.904
53.275
46.016
86.022
191.058
64.337
53.926
36.215
86.384
195.084
67.120
53.600
46.880
90.671
197.083
75.050
58.360
53.096
95.897
Average
191.279
67.353
54.790
45.552
Note: A1= rregular sze, A2= 1 x 5 cm, A3= 3 x 5 cm, A4= 5 x 5 cm.
Table 3. CPH theobromne content KBK nonfermented and fermented wth A. ficuum (ppm)
Chop szes
Incubaton
tmes (days)
A1
A2
A3
A4
0
5
7
9
Average
175.409
68.566
62.146
45.052
87.793
156.430
69.884
66.348
42.914
83.894
214.776
81.920
75.749
58.360
107.701
223.609
91.207
88.566
80.048
120.858
Average
192.556
77.894
73.202
56.593
Note: A1= rregular sze, A2= 1 x 5 cm, A3= 3 x 5 cm, A4= 5 x 5 cm.
average of theobromne content was acheved through fermentaton at 9 days of
ncubaton (decreased 28.09%). From ths experment, the lowest of theobromne
content (36.512 ppm) was acheved n A2 chop sze when ncubated durng 9 days.
The average for theobromne content n CPH non fermentaton (0 day) was
192.556 ppm (192.556 mg/kg), kg). Ths value was lower from reported by Haryat
and Harjosuwto (1984); Wong and Osman (1988) about 0.17-0.20% (1,700-2,000
mg/kg), but EFSA (2008) reported hgher reach 0.40% (4,000 mg/kg). Dfferent
of theobromne content for non
on fermented CPH was caused by dfferent treatment
of chop sze.. Ths research, CPH was chopped and ar dred durng 8 hours whch
was decrease water content and followed by decreasng of theobromne content..
The cocoa bean had theobromne content hgher reached 1.5-3.0%
.5-3.0%
5-3.0%
.0%
% (15,000-30,000
mg/kg)) (Harrson,
Harrson, 2001), whle dry beans had theobromne content lower only 1.8%
(18,000 mg/kg) (Wllson, 1999).. The CPH theobromne content was the lowest
among 3 varetes of Aspergillus spp. after fermentaton process that was A. niger n
A2 wth ncubaton of 9 days (45.052 ppm).
Reducng of CPH theobromne content caused by decomposng actvty of
Aspergillus spp. whch was durng fermentaton process to result heat (ncreasng
178
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
of substrate temperature). Besde that occured hydrolyss n CPH substrate. In ths
step, theobromne dssolve n cell lqud and came out together wth evaporaton
process. Rohan (1963) and Alamsyah (1991) were reported that theobromne content reduce durng fermentaton process because soluble n cell lqud and dffuson n cocoa bean nb. Dffuson process wll stop f occur balance of theobromne
content n cocoa bean.. Decreasng of CPH theobromne content was relatvely low
between A. niger compared A. ficuum. Ths case had relatton wth ablty A. niger
to utlze theobromne as energy source. Accordng to Asano et al. (1993) and Hakl
et al. (1999) that A. niger had ablty to utlze theobromne as energy source through methylxanthnes degradaton.
Conclusons
The chop sze of 1x5 cm and ncubaton tmes of 9 days affected decreasng of
CPH theobromne content. The best of CPH fermented
ermented for reducng of theobromne
content for chop sze 1x5 cm (A2) wth 9 days fermentaton, fermented by A.
oryzae.
References
Alamsyah, T.S. 1991. Peranan fermentas dalam pengolahan bj kakao kerng.
Berta Peneltan Perkebunan. 1 (2): 97-103.
Aregheore, E.M. 1999. Ant-qualty and toxc components n some foods consumed
by humans and lvestock n the South Pacfc regon. Revew PNG. J. of Agrc.
Forestry and Fsheres. 42: 15-21.
Asano, Y., T. Komeda and H. Yamada. 1993. Mcrobal producton of theobromne
from caffen. Bosc. Botech. Bochem. 57: 1286-1289.
European Food Safety Authorty. 2008. Theobromne as undesrable substances n
anmal feed. Scentfc Opnon of The Panel on Contamnatons n The Food
Chan. The EFSA J. 752:1-66.
Goldng, E.J. 1985. Provdng Energy-Proten Supplementaton durng The Dry
Season. In: Nutrton of Grazng Rumnants n Warm Clmates. Edted by
L.R. McDowell. Academc Press, Inc. Orlando, Florda, USA. pp. 130-163.
30-163.
-163.
63.
Hakl, M., F. Vosnet, G.V. Gonzales and C. Augur. 1999. Caffene degradaton n
sold-state fermentaton by Aspergillus tamari:
Harrson, K. 2001. Theobromne. Theobromine@3Dchem.com
@3Dchem.com Update by K. Harrison (Molecule of the Month for February 2001). http://www.3dchem.com/
molecules.asp?ID=155#. Dakses tanggal 9 September 2008.
Haryat, T. dan B. Harjosuwto. 1984. Pemanfaatan Lmbah Coklat sebaga Bahan
Dasar Pembuatan Pektn. Menara Perkebunan, Jember.
Huq, F. 2006. Moleculer modellng analyss of the metabolsm of caffen. Asan J.
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
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Bochem. 1: 276-286.
Lubs, D., E. Wna, B. Haryanto dan T. Suhargyantatmo. 2002a. Effectveness of
Aspergillus oryzae fermentaton to mprove dgeston of fbrous feeds:
In
Vitro. JITV. 7 (2): 90-98.
Mahyuddn, P. and B. Bakre. 1993. Defferent level of cocoa shell n dets of growng cattle. Ilmu dan Peternakan. 6 (2): 1-4.
Rohan, T.A. 1963. Processng of Raw Cocoa for The Market. FAO. Agrc. Studes.
No. 60, Rome.
Urch, K. 1994. Comparatve Anmal Bochemstry. @ Sprnger-Verlag, Berln Hedelberg, Germany.
Wllson, K.C. 1999. Crop Producton Scence n Hortculture: Coffee, Cocoa and
Tea. CABI Publshng, Unted Kngdom by The Unversty Press, Cambrdge,
London. pp. 100-165.
Wong, H.K., and A.H. Osman. 1988. Nutrtve value and rumen fermentaton profle of sheep fed of fresh or dred cocoa pod husk based dets. J. of MARDI
Res. 16 (2): 147-156.
Yamaoka-Yano, D.M. and P. Mazzafera. 1999. Catabolc pathway of caffene and
purfcaton of a xanthne oxdase responsble for mathylurc acd producton
n Pseudomonas putida L. Revista de Microbiologia. 30: 70-78.
180
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
(
cacao)
Fermented by Aspergillus spp. in Different of Chop Sizes and
Fermentation Times
F. F. Munier1 & H. Hartadi2
1
Graduate Student of Animal Science Institute, Gadjah Mada University, Fauna
Street No. 3, Bulaksumur, Yogyakarta, 55281
2
Animal Science Faculty, Gadjah Mada University, Fauna Street No. 3,
Bulaksumur, Yogyakarta, 55281
Abstract
Theobromine is one of alkaloid in cocoa pod husk (CPH). Fresh CPH has
high theobromine content reach 0.40% (400.000 ppm) which is fresh CPH portion
in ruminant ration more than 30% to be affected of health. This research aim was
to know the change of theobromine content of CPH with different chop sizes were
fermented in the different time of incubation by Aspergillus spp. Three species of
Aspergillus spp. were used in this research i.e. A. niger, A. oryzae and A. ficuum
BPT. There were 4 chop sizes i.e.; irrigular size (A1), 1x5 cm (A2), 3x5 cm (A3), and
5x5 cm (A4). The incubation times were; 0, 5, 7, and 9 days, with three replications.
The solely parameter was theobromine content which were prepared according to
AOAC (1990) and analyze using HPLC (Chen et al., 2008). The result of research
showed that CPH fermented using A. niger, the lowest of theobromine content for
chop size A2 and incubated 9 days was 38.088 ppm. CPH fermented using A. oryzae
had the lowest of theobromine content in A2 and 9 days only 36.215 ppm. CPH
fermented with A. ficuum had the lowest of theobromine content in A2 and 9 days
reach 42.914 ppm. It was concluded that the best of CPH fermented
ermented for reducing of
theobromine content for 1x5 cm (A2) chop size with 9 days fermentation, fermented
by A. oryzae.
Keywords: theobromine,, cocoa pod husk, Aspergillus spp., chop sizes, incubation
times
Introducton
Cocoa pod husk (CPH) utlzaton s expected to be capable of feed fulfllng the
rumnant anmal requrement yearly, so the anmal wll not be lackng of nutrents for
ts lfe and producton. Ths condton has reason because cocoa trees have frut n
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
175
a long year whch s CPH abundant avalable n harvest season. Utlzaton of fresh
CPH as feed supplement has lmtng factor whch one alkalod s theobromne. CPH
has hgh of theobromne content or 3,7-dmethylxanthne (Mahyuddn and Bakre,
1993; Aregheore, 1999).
). The hgh concentraton theobromne affected the health of
anmal (Goldng, 1985). The theobromne content of CPH about 0.17-0.20% (Wong
and Hassan, 1988),
8),
), whle n cocoa beans reached 1.5-3.0%,
.5-3.0%,
5-3.0%,
.0%,
%, and n dry cocoa beans
reached 1.8%
.8% (Wllson, 1999)..
Decreasng of theobromne content can be done wth fermentaton method usng mcrobe agent such as fung. Some researchers used A. niger as decomposer
of CPH n sold fermentaton process (Yamaoka-Yano and Mazzafera, 1999; Huq,
2006) or A.. oryzae (Lubs et al., 2002). They found that CPH theobromne content
were reduced due to the role of enzyme actvty that secreted by Aspergillus spp.
Based on those results, n ths research we used A. niger, A. oryzae and A. ficuum n sold fermentaton wth CPH as a substrate. The am of ths research was
to decrease theobromne content of CPH n Laboratory scale that subsequently the
fermented CPH would used for alternatve forage, and also we studed about storage
tme of fermented CPH as feed.
Materals and Methods
Ths experment was fermented CPH usng three of Aspergillus spp..e. A. niger, A. oryzae and A. ficuum, n four types of chop szes .e. rregular sze (A1) as
control, 1 x 5 cm (A2), 3 x 5 cm (A3), and 5 x 5 cm (A4) and four knds of ncubaton
tmes .e. 0, 5, 7 and 9 days. CPH was ar dred for 8 hours after choppng, whle
powder fung were used as much as 1% and then added urea 1% of CPH weght.
Urea dssolved wth sterle water n rato 1 : 10 parts. Fresh CPH used 1 kg every
treatment whch ths was spread n plastc box (the frst coatng). CPH sprayed wth
urea and noculated of Aspergillus spp., the next wth same procedure was done for
the second and thrd coatngs.
After completon of fermentaton process, CPH was removed and to be ardred for 6 hours. The weghng of CPH and to be contnued wth samples dryng
usng oven n temperature of 55 oC for 4 days. About 100 g of fermented CPH was
dred usng freeze dryer for 4 days. Dred CPH was ground wth the mortal whch
was surrounded by ce to mantan optmal temperature and theobromne content,
and then followed by analyss of theobromne content of CPH. Analyss of theobromne content was recommended by EFSA (2008) usng Hgh-Performance Lqud
Chromatography (HPLC).
Results and Dscusson
Theobromne as alkalod compound cause btter n cocoa beans and CPH. Al-
176
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
kalod s organc component from plant whch has contan of ntrogen, commonly as
alkals and t has bology actvty. The alkalod n plant has functon for attact of pest
(Urch, 1994). Fermentas treatment can decrease CPH theobromne content. CPH
non fermented has hgher of theobromne content (Tablee 1).
).
Table 1. Theobromne content of cocoa pod husk fermented by A. niger (ppm)
Treatment of chop szes
Incubaton
tmes (days)
A1
A2
A3
A4
0
5
7
9
Average
157.560
76.008
38.545
38.477
77.648
134.410
71.093
38.103
38.088
70.424
171.554
78.857
46.680
43.837
85.232
186.193
81.778
52.697
45.727
91.599
Average
162.429
76.934
44.006
37.507
Note: A1= rregular sze, A2= 1 x 5 cm, A3= 3 x 5 cm, A4= 5 x 5 cm.
The fndng results showed that fermentaton process affected the decreasng
of CPH theobromne content. In A. niger fermentaton treatment, there was an effect
on CPH theobromne content due to dfferences of chop szes and ncubaton tmes.
Reducng of CPH chop sze was parallel to reducng of theobromne content. The
A2 and A4 chop szes showed average of the lowest and the hghest of theobromne
content, respectvely. On the other sde, ncreasng of ncubaton tme was nversed
wth decreasng of average of theobromne content. The lowest average of theobromne content was acheved through fermentaton at 9 days of ncubaton (reduced
76.91%). From ths experment, the lowest of theobromne content (38.088 ppm)
was acheved n A2 chop sze when ncubated durng 9 days.
Fermentaton
ermentaton usng A. oryzae n treatment of chop szes and ncbaton tmes
showed effect to decreasng of CPH theobromne content that was presented n Table 2. Reducng of CPH chop sze had correlaton wth reducng of theobromne
content. The A2 chop sze showed average of the lowest theobromne content, whle
A4 was the hghest theobromne content. The longer ncubaton tme supported reducng theobromne content. The lowest average of theobromne content was occur
at 9 days fermentaton (reduced 23.81%). From ths experment, the lowest of theobromne content (36.215 ppm) was acheved n A2 chop sze when ncubated durng
9 days.
The fndng results showed that fermentaton CPH usng A. ficuum affected
the decreasng of theobromne content (Table
Table 3). Decreasng of CPH chop sze had
relaton wth decreasng of theobromne content. The A2 chop sze showed average
of the lowest for theobromne content. On the other sde, ncreasng of ncubaton
tme was nversed wth decreasng of average of theobromne content. The lowest
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
177
Table 2. CPH theobromne content KBK nonfermented and fermented wth A. niger (ppm)
Chop szes
Incubaton
tmes (days)
A1
A2
A3
A4
0
5
7
9
Average
181.891
62.904
53.275
46.016
86.022
191.058
64.337
53.926
36.215
86.384
195.084
67.120
53.600
46.880
90.671
197.083
75.050
58.360
53.096
95.897
Average
191.279
67.353
54.790
45.552
Note: A1= rregular sze, A2= 1 x 5 cm, A3= 3 x 5 cm, A4= 5 x 5 cm.
Table 3. CPH theobromne content KBK nonfermented and fermented wth A. ficuum (ppm)
Chop szes
Incubaton
tmes (days)
A1
A2
A3
A4
0
5
7
9
Average
175.409
68.566
62.146
45.052
87.793
156.430
69.884
66.348
42.914
83.894
214.776
81.920
75.749
58.360
107.701
223.609
91.207
88.566
80.048
120.858
Average
192.556
77.894
73.202
56.593
Note: A1= rregular sze, A2= 1 x 5 cm, A3= 3 x 5 cm, A4= 5 x 5 cm.
average of theobromne content was acheved through fermentaton at 9 days of
ncubaton (decreased 28.09%). From ths experment, the lowest of theobromne
content (36.512 ppm) was acheved n A2 chop sze when ncubated durng 9 days.
The average for theobromne content n CPH non fermentaton (0 day) was
192.556 ppm (192.556 mg/kg), kg). Ths value was lower from reported by Haryat
and Harjosuwto (1984); Wong and Osman (1988) about 0.17-0.20% (1,700-2,000
mg/kg), but EFSA (2008) reported hgher reach 0.40% (4,000 mg/kg). Dfferent
of theobromne content for non
on fermented CPH was caused by dfferent treatment
of chop sze.. Ths research, CPH was chopped and ar dred durng 8 hours whch
was decrease water content and followed by decreasng of theobromne content..
The cocoa bean had theobromne content hgher reached 1.5-3.0%
.5-3.0%
5-3.0%
.0%
% (15,000-30,000
mg/kg)) (Harrson,
Harrson, 2001), whle dry beans had theobromne content lower only 1.8%
(18,000 mg/kg) (Wllson, 1999).. The CPH theobromne content was the lowest
among 3 varetes of Aspergillus spp. after fermentaton process that was A. niger n
A2 wth ncubaton of 9 days (45.052 ppm).
Reducng of CPH theobromne content caused by decomposng actvty of
Aspergillus spp. whch was durng fermentaton process to result heat (ncreasng
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Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
of substrate temperature). Besde that occured hydrolyss n CPH substrate. In ths
step, theobromne dssolve n cell lqud and came out together wth evaporaton
process. Rohan (1963) and Alamsyah (1991) were reported that theobromne content reduce durng fermentaton process because soluble n cell lqud and dffuson n cocoa bean nb. Dffuson process wll stop f occur balance of theobromne
content n cocoa bean.. Decreasng of CPH theobromne content was relatvely low
between A. niger compared A. ficuum. Ths case had relatton wth ablty A. niger
to utlze theobromne as energy source. Accordng to Asano et al. (1993) and Hakl
et al. (1999) that A. niger had ablty to utlze theobromne as energy source through methylxanthnes degradaton.
Conclusons
The chop sze of 1x5 cm and ncubaton tmes of 9 days affected decreasng of
CPH theobromne content. The best of CPH fermented
ermented for reducng of theobromne
content for chop sze 1x5 cm (A2) wth 9 days fermentaton, fermented by A.
oryzae.
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