Biodegradation of coffee husk substrate during the mycelia growth of Pleurotus ostreatus and the effect on in vitro digestibility
Biodegradation of coffee husk substrate during the mycelia growth
of Pleurotus ostreatus and the effect on in vitro digestibility
Irma Badarina1, D. Evvyernie1, T. Toharmat1, E. N. Herliyana2& L.K. Darusman3
1
Department of Nutrition and Feed Technology, Faculty of Animal Science, Bogor
Agricultural University, Bogor 16680 Indonesia,
Email: erniedwierra@yahoo.com
2
Department of Silviculture,Faculty of Forestry, Bogor Agricultural University
3
Departement of Chemistry, Faculty of Mathematics and Natural Science, Bogor
Agricultural University.
Abstract
The aim of this studies were conducted to evaluate culturing of mushroom
P.ostreatus on coffee husk in solid state fermentation as means of improving the
nutritive value of coffee husk for ruminant animals. The influence of P.ostreatus
on coffee husk biodegradation was investigated. The dry matter and composition
changes of coffee husk substrate for P.ostreatus cultivation were analysed on day
0, 30 and 60 after seeding. The profile of cellulose, hemicellulose and lignin were
changed when it was used by P.ostreatus. Meanwhile their rate of change varied
at different growing day. The increase of protein content and the reduction of
lignocellulose content increase dry matter digestibility of coffee husk substrate.
This fact could provide an alternative of biofermentation product based on coffee
husk substrate which is safe for environment.
Keywords: biodegradation, coffee husk, digestibility, substrate, P. ostreatustion
Introducton
Pleurotus ostreatus s one of the popular cultvated mushroom. It can be
cultvated on a wde range of lgnoselulosc substrates such as wheat straw, cocoa
husk and cotton stalks (Fazael et al., 2004; L et al., 2001; Alemawor, 2009).
Pleurotus ostreatus belongs to whte rot fung whch are able to degrade lgnn because
produce lgnnolytc extracellular enzymes, such as laccases, lgnn peroxdases and
Mn peroxdases (Kerem et al., 1992; Chang and Mles, 2004).
The ablty of P.ostreatus degrades a wde varety of lgnoselulosc substrates,
enablng t to play an mportant role n managng organc wastes whose dsposal s
problematc. Pleurotus speces have been used by human for ther nutrtonal value,
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
341
medcnal propertes, transformaton of wastes nto anmal feed and other benefcal
effects (Hadar and Araz, 1986; Gregor et al. 2007; Adamovc et al., 1998).
Coffee husk s the major byproducts produced durng the operaton of coffee
cherry to get coffee gran by sun dryng (Fan et al., 2004). In coffee-producng regons, coffee husk s barely utlzed. Therefore, t s consdered the most abundant
pollutant materal. Coffee husk has potency as a source of rumnant feed. The proten content s 9.2-11.3% equally to rce bran proten (±10.4%) and has metabolc
energy around 3.356 Kcal/kg (Zanudn and Murtsar, 1995). The content of lgnn
s 35.0-40.0% (Fan and Soccol, 2005). The dgestblty of these materals are lmted by the presence of lgnn whch prevents access of hydrolytc enzymes to cellulose and hemcelluloses.
Applcaton of Pleurotus ostreatus s worth consderng for mprovng the nutrtve value of coffee husk. Ths study was carred out to asses the effect of a sold
state fermentaton nvolvng Pleurotus ostreatus on the nutrton composton of coffee husk and to evaluate n vtro dgestblty. In addton, fermentaton perod on the
process was evaluated.
Materals and Methods
Coffee husk were obtaned from coffee hullng plant at Rejang Lebong
Resdence Bengkulu Provnce. Coffee husks were ar-dred to mosture content 1015%. The sold state substrate were prepared wth the composton adopted from
sawdust standard medum (Herlyana et al. 2008). The mushroom substrate may be
defned as a knd of lgnocellulosc materal supports the growth, development and
frutng of mushroom. The substrate were conssted of 82,5% coffee husk, 15% rce
bran, 1,5%gps and 1,0% CaCO3. The clean water were added to the substrate as
much as 60-65% (v/w). All these components were placed n polypropylene bag n
amount 400 gram per bag. Each bag was closed wth a small cotton plug nserted n
the mddle of ts openng. The bags were sterlzed at 121oC for 30 mnutes. After
cool, each of bag was seeded wth 15 gram (3,75%) of Pleurotus ostreatus spawn.
All spawned bag were placed n growng room wth the temperature was 22-28oC
and relatve humdty 60-80%. After 30 days, the substrate was fully colonzed, and
on 60 days prmordal started to appeared.
The content of proten was analyzed usng Kjeldahl method. The cell wall
components (NDF, ADF, Lgnn, cellulose and hemcelluloses) were analyze usng
deterjent analyze method as descrbed by Goerng and Van Soest (1970). In vitro dry
matter dgestblty was evaluated accordng to Tlley and Terry method (1963).
The treatment was the fermentaton tme conssted of 0 untreated), 30 and 60
days after seedng. The nutrent composton changes were descrbed descrptvely.
For the dry matter measurement the treatment was arranged n Block Randomsed
Desgn (3x4). The rumen noculum were obtaned from four cattles as block.
342
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
Sgnfcant dfferences were calculated usng Duncan’s multple range test followng
analyss of varance.
Result and dscusson
The celluloses, hemcelluloses and lgnn are the man sources of carbon and
energy for P.ostreatus growth, whle proten serves as the ntrogen source. Ther
degradaton and utlzaton can greatly affect P.ostreatus growth and resultng feed
value of the substrate. The change of nutrent composton contents durng the
P.ostreatus mycela growth perod are shown n Table 1.
There were ncreasng of proten content and decreasng of fber fracton (lgnn,
NDF and ADF) produced by bofermentaton. The decreasng of fber fracton s the
ndcaton that Pleurotus ostreatus can degrade the cell wall component of coffee
husk.
The decreasng of NDF and ADF from coffee husk suggested that these fung
could utlze the cell wall component as carbon source and energy for growth. The
decreasng of NDF and ADF contents of treated coffee byproduct has been reported
by Penaloza et al., (1985). The decreasng of NDF, ADF and ADL n the frst 30 days
of mycela growth were 2.339%, 4.586% and 19.874%, respectvely. Meanwhle
n 60 days, the decreasng of NDF, ADF and ADL were 16.587%, 15.036% and
31.161%, respectvely from the ntal value.
The fermentaton tme was mportant to mprove the nutrtve value of straw.
The longer fermentaton perod led greater depleton of carbohydrate source of
coffee husk by fung. Ths condton could mprove the dgestblty of coffee husk as
result of the changes n non structural carbohydrate to structural carbohydrate rato.
Decreasng of lgnn n coffee husk could be a result of lgnn degradng enzymes
produced by Pleurotus (Hong et al., 2003). These result are supported by the report
from Wdastut et al. (2008) who noted lgnnoltyc enzyme actvtes followed the
pattern of lgnn dsappearance from substrate and drectly corrected wth tme of ts
dsappearance. Plat and Hadar (1983) noted that durng the mycela growth perod,
P.ostreatus mycela were more capable to degrade lgnn, and the degradaton of
lgnn played an mportant role n mycela development.
The rapd decreasng of hemcellulosc component n 30 days fermentaton
showed that hemcelluloses were the frst substrate utlzed by mycela as the carbon
and energy sources at the begnnng phase of growth. The decreasng of hemcelluloses was 31,578% from ntal value n 30 days fermentaton. Ths suggest that
hemcellulose s more easly degraded than cellulose and lgnn. Pleurotus ostreatus
mushroom secreted enzym to demolsh the easer used compound. Pleurotus ostreatus needs a carbon source whch s easer to metabolze (Crawford, 1981). Hemcelluloses were degraded easer than cellulose and lgnn (Perez, 2002).
The cellulose content ncreased 35.574% n 30 days and 27.063% n 60 days.
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
343
Bofermentaton broke the lgnocelluloses bond. Delgnfcaton has mportant
role n mycela growth whch cleavage polysaccharde component (cellulose and
hemcelluloses) (Agosn and Oder, 1985). Ths component wll be utlzed by fung
as substrate for ther growth (Hatakka, 2004).
Durng the mycela growth, the proten content ncreased 0.927% n 30 days
and 17.220% n 60 day fermentaton. Mycela n 60 days were thcker than 30 days.
Fungal cell n mycela contrbuted the proten content of subtrate because 60 and
70% of ntrogen present n the fungal cell s proten (Chang and Mles 2004). The
hgher proten content n 60 days n the substrate were prepared to transferable
ntrogen nto frut bodes. The extensve formaton of prmorda n 60 days ndcated
the end of the vegetatve growth phase of P.ostreatus. As coffee husk substrate
was degraded and nutrent used by P.ostreatus, the total organc matter of substrate
decreased (Table 1).
The ncreasng of proten content and the decreasng of lgnocelluloses of
coffee husk after fermentaton showed that Coffee husk could be used as substrate
P.ostreatus cultvaton. The mprovng nutrton value after fermentaton especally
on 60 days ndcated that the substrate can be used as a product feed.
In vitro dry matter dgestblty tests for rumnant were conducted for the
dgestblty of untreated and treated coffee husk. Four replcaton were conducted
and the result are shown n fgure 1. Average dry matter dgestblty (Table 1) ncreased sgnfcantly 4.983% n 60 days fermentaton and decreased 14.435% n 30
days fermentaton from untreated coffee husk. The possblty of ths condton s
that n 30 days fermentaton the hgher level of cellulose made dgestblty lower.
Table 1. Changes of nutrent contents and average in vitro dry matter dgestblty of coffee
husk substrate durng Pleurotus ostreatus mycela growng (0, 30, and 60 days
fermentaton) (as % dry matter)
Nutrent contens
(%)
Organc matter
Crude Proten
NDF
ADF
Hemcelluloses
Cellulose
Lgnn
Dry Matter
Dgestblty (%)
0 days (Untreated)
(Treated) 30 days
after seedng
(Treated) 60 days
after seedng
93.710
10.360
95.177
87.184
7.993
19.514
65.421
29.518±1.249a
92.950
10.456
92.950
83.186
5.469
26.456
52.419
25.257±0.721b
86.599
12.144
79.390
74.075
5.3170
24.795
45.035
30.989±1.263c
Dfferent superscrpt n the same row means sgnfcantly dfferent (P
of Pleurotus ostreatus and the effect on in vitro digestibility
Irma Badarina1, D. Evvyernie1, T. Toharmat1, E. N. Herliyana2& L.K. Darusman3
1
Department of Nutrition and Feed Technology, Faculty of Animal Science, Bogor
Agricultural University, Bogor 16680 Indonesia,
Email: erniedwierra@yahoo.com
2
Department of Silviculture,Faculty of Forestry, Bogor Agricultural University
3
Departement of Chemistry, Faculty of Mathematics and Natural Science, Bogor
Agricultural University.
Abstract
The aim of this studies were conducted to evaluate culturing of mushroom
P.ostreatus on coffee husk in solid state fermentation as means of improving the
nutritive value of coffee husk for ruminant animals. The influence of P.ostreatus
on coffee husk biodegradation was investigated. The dry matter and composition
changes of coffee husk substrate for P.ostreatus cultivation were analysed on day
0, 30 and 60 after seeding. The profile of cellulose, hemicellulose and lignin were
changed when it was used by P.ostreatus. Meanwhile their rate of change varied
at different growing day. The increase of protein content and the reduction of
lignocellulose content increase dry matter digestibility of coffee husk substrate.
This fact could provide an alternative of biofermentation product based on coffee
husk substrate which is safe for environment.
Keywords: biodegradation, coffee husk, digestibility, substrate, P. ostreatustion
Introducton
Pleurotus ostreatus s one of the popular cultvated mushroom. It can be
cultvated on a wde range of lgnoselulosc substrates such as wheat straw, cocoa
husk and cotton stalks (Fazael et al., 2004; L et al., 2001; Alemawor, 2009).
Pleurotus ostreatus belongs to whte rot fung whch are able to degrade lgnn because
produce lgnnolytc extracellular enzymes, such as laccases, lgnn peroxdases and
Mn peroxdases (Kerem et al., 1992; Chang and Mles, 2004).
The ablty of P.ostreatus degrades a wde varety of lgnoselulosc substrates,
enablng t to play an mportant role n managng organc wastes whose dsposal s
problematc. Pleurotus speces have been used by human for ther nutrtonal value,
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
341
medcnal propertes, transformaton of wastes nto anmal feed and other benefcal
effects (Hadar and Araz, 1986; Gregor et al. 2007; Adamovc et al., 1998).
Coffee husk s the major byproducts produced durng the operaton of coffee
cherry to get coffee gran by sun dryng (Fan et al., 2004). In coffee-producng regons, coffee husk s barely utlzed. Therefore, t s consdered the most abundant
pollutant materal. Coffee husk has potency as a source of rumnant feed. The proten content s 9.2-11.3% equally to rce bran proten (±10.4%) and has metabolc
energy around 3.356 Kcal/kg (Zanudn and Murtsar, 1995). The content of lgnn
s 35.0-40.0% (Fan and Soccol, 2005). The dgestblty of these materals are lmted by the presence of lgnn whch prevents access of hydrolytc enzymes to cellulose and hemcelluloses.
Applcaton of Pleurotus ostreatus s worth consderng for mprovng the nutrtve value of coffee husk. Ths study was carred out to asses the effect of a sold
state fermentaton nvolvng Pleurotus ostreatus on the nutrton composton of coffee husk and to evaluate n vtro dgestblty. In addton, fermentaton perod on the
process was evaluated.
Materals and Methods
Coffee husk were obtaned from coffee hullng plant at Rejang Lebong
Resdence Bengkulu Provnce. Coffee husks were ar-dred to mosture content 1015%. The sold state substrate were prepared wth the composton adopted from
sawdust standard medum (Herlyana et al. 2008). The mushroom substrate may be
defned as a knd of lgnocellulosc materal supports the growth, development and
frutng of mushroom. The substrate were conssted of 82,5% coffee husk, 15% rce
bran, 1,5%gps and 1,0% CaCO3. The clean water were added to the substrate as
much as 60-65% (v/w). All these components were placed n polypropylene bag n
amount 400 gram per bag. Each bag was closed wth a small cotton plug nserted n
the mddle of ts openng. The bags were sterlzed at 121oC for 30 mnutes. After
cool, each of bag was seeded wth 15 gram (3,75%) of Pleurotus ostreatus spawn.
All spawned bag were placed n growng room wth the temperature was 22-28oC
and relatve humdty 60-80%. After 30 days, the substrate was fully colonzed, and
on 60 days prmordal started to appeared.
The content of proten was analyzed usng Kjeldahl method. The cell wall
components (NDF, ADF, Lgnn, cellulose and hemcelluloses) were analyze usng
deterjent analyze method as descrbed by Goerng and Van Soest (1970). In vitro dry
matter dgestblty was evaluated accordng to Tlley and Terry method (1963).
The treatment was the fermentaton tme conssted of 0 untreated), 30 and 60
days after seedng. The nutrent composton changes were descrbed descrptvely.
For the dry matter measurement the treatment was arranged n Block Randomsed
Desgn (3x4). The rumen noculum were obtaned from four cattles as block.
342
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
Sgnfcant dfferences were calculated usng Duncan’s multple range test followng
analyss of varance.
Result and dscusson
The celluloses, hemcelluloses and lgnn are the man sources of carbon and
energy for P.ostreatus growth, whle proten serves as the ntrogen source. Ther
degradaton and utlzaton can greatly affect P.ostreatus growth and resultng feed
value of the substrate. The change of nutrent composton contents durng the
P.ostreatus mycela growth perod are shown n Table 1.
There were ncreasng of proten content and decreasng of fber fracton (lgnn,
NDF and ADF) produced by bofermentaton. The decreasng of fber fracton s the
ndcaton that Pleurotus ostreatus can degrade the cell wall component of coffee
husk.
The decreasng of NDF and ADF from coffee husk suggested that these fung
could utlze the cell wall component as carbon source and energy for growth. The
decreasng of NDF and ADF contents of treated coffee byproduct has been reported
by Penaloza et al., (1985). The decreasng of NDF, ADF and ADL n the frst 30 days
of mycela growth were 2.339%, 4.586% and 19.874%, respectvely. Meanwhle
n 60 days, the decreasng of NDF, ADF and ADL were 16.587%, 15.036% and
31.161%, respectvely from the ntal value.
The fermentaton tme was mportant to mprove the nutrtve value of straw.
The longer fermentaton perod led greater depleton of carbohydrate source of
coffee husk by fung. Ths condton could mprove the dgestblty of coffee husk as
result of the changes n non structural carbohydrate to structural carbohydrate rato.
Decreasng of lgnn n coffee husk could be a result of lgnn degradng enzymes
produced by Pleurotus (Hong et al., 2003). These result are supported by the report
from Wdastut et al. (2008) who noted lgnnoltyc enzyme actvtes followed the
pattern of lgnn dsappearance from substrate and drectly corrected wth tme of ts
dsappearance. Plat and Hadar (1983) noted that durng the mycela growth perod,
P.ostreatus mycela were more capable to degrade lgnn, and the degradaton of
lgnn played an mportant role n mycela development.
The rapd decreasng of hemcellulosc component n 30 days fermentaton
showed that hemcelluloses were the frst substrate utlzed by mycela as the carbon
and energy sources at the begnnng phase of growth. The decreasng of hemcelluloses was 31,578% from ntal value n 30 days fermentaton. Ths suggest that
hemcellulose s more easly degraded than cellulose and lgnn. Pleurotus ostreatus
mushroom secreted enzym to demolsh the easer used compound. Pleurotus ostreatus needs a carbon source whch s easer to metabolze (Crawford, 1981). Hemcelluloses were degraded easer than cellulose and lgnn (Perez, 2002).
The cellulose content ncreased 35.574% n 30 days and 27.063% n 60 days.
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
343
Bofermentaton broke the lgnocelluloses bond. Delgnfcaton has mportant
role n mycela growth whch cleavage polysaccharde component (cellulose and
hemcelluloses) (Agosn and Oder, 1985). Ths component wll be utlzed by fung
as substrate for ther growth (Hatakka, 2004).
Durng the mycela growth, the proten content ncreased 0.927% n 30 days
and 17.220% n 60 day fermentaton. Mycela n 60 days were thcker than 30 days.
Fungal cell n mycela contrbuted the proten content of subtrate because 60 and
70% of ntrogen present n the fungal cell s proten (Chang and Mles 2004). The
hgher proten content n 60 days n the substrate were prepared to transferable
ntrogen nto frut bodes. The extensve formaton of prmorda n 60 days ndcated
the end of the vegetatve growth phase of P.ostreatus. As coffee husk substrate
was degraded and nutrent used by P.ostreatus, the total organc matter of substrate
decreased (Table 1).
The ncreasng of proten content and the decreasng of lgnocelluloses of
coffee husk after fermentaton showed that Coffee husk could be used as substrate
P.ostreatus cultvaton. The mprovng nutrton value after fermentaton especally
on 60 days ndcated that the substrate can be used as a product feed.
In vitro dry matter dgestblty tests for rumnant were conducted for the
dgestblty of untreated and treated coffee husk. Four replcaton were conducted
and the result are shown n fgure 1. Average dry matter dgestblty (Table 1) ncreased sgnfcantly 4.983% n 60 days fermentaton and decreased 14.435% n 30
days fermentaton from untreated coffee husk. The possblty of ths condton s
that n 30 days fermentaton the hgher level of cellulose made dgestblty lower.
Table 1. Changes of nutrent contents and average in vitro dry matter dgestblty of coffee
husk substrate durng Pleurotus ostreatus mycela growng (0, 30, and 60 days
fermentaton) (as % dry matter)
Nutrent contens
(%)
Organc matter
Crude Proten
NDF
ADF
Hemcelluloses
Cellulose
Lgnn
Dry Matter
Dgestblty (%)
0 days (Untreated)
(Treated) 30 days
after seedng
(Treated) 60 days
after seedng
93.710
10.360
95.177
87.184
7.993
19.514
65.421
29.518±1.249a
92.950
10.456
92.950
83.186
5.469
26.456
52.419
25.257±0.721b
86.599
12.144
79.390
74.075
5.3170
24.795
45.035
30.989±1.263c
Dfferent superscrpt n the same row means sgnfcantly dfferent (P