Emirates Journal of Food and Agriculture. 2016.

28(3): 188-195
doi: EJFA-2015-05-192
http://www.ejfa.me/

REGULAR ARTICLE

Introduction of actinomycetes starter on coffee fruits

fermentation to enhance quality of coffee pulp
Nurleni Kurniawati1, Anja Meryandini2, Titi Candra Sunarti3*
Study Program of Biotechnology, Bogor Agricultural University, Bogor, Indonesia, 2Department of Agroindustrial Technology, Bogor Agricultural
1

University, Bogor, Indonesia, 3Department of Biology, Bogor Agricultural University, Bogor, Indonesia

ABSTRACT
Coffee pulp contains bioactive compounds such as polyphenols and aldehydes, which are covalently linked to the pulp cell wall. The research
aimed to determine of potential Actinomycetes concortia on the degradation of coffee pulp to enhance the yield and quality of coffee
pulp polyphenol extracts. In this study, whole coffee fruit were fermented in solid-state cultivation by using concortia of Streptomyces
exfoliatus 42 and Streptomyces costaricanus 45I-3 that having cellulolytic and xylanolytic activities. The introduction of Actinomycetes
starter accelerated the fermentation which caused the degradation of lignocellulosic components of coffee pulp, and significantly effects
on the yield of bioactive compounds such as polyphenols, anthocyanins, tannins, and catechin. Compared to the spontaneous fermentation
of coffee fruit, the highest yield of bioactive components extracts produced from 6th day incubation for total polyphenols (1.20 mg mL-1),
anthocyanin (109.95 mg mL-1) and the highest yield of catechin (10.38 mg mL-1) produced form 3th day incubation, but introduction of
Actinomycetes reduced the tannin contents after fermentation.
Keywords: Coffee pulp; Polyphenol; Actinomycetes concortia; Anthocyanin; Tannin

INTRODUCTION coffee bean can produce variety in quality of coffee

products. The bacteria and yeast cultures are usually
Indonesia is one of biggest producer and exporter of introduced to make homogenous condition of spontaneous
coffee. Indonesia coffee plantation covers 1.305.895
​​ ha fermentation process. The used of bacterial starter culture
of land area; and produces 748.109 tonnes of dried coffee in fermentation process could reduce the fermentation time
bean per year (Ditjenbun, 2012). Coffee fruit cannot be and improve coffee bean quality (Silva et al., 2013). Specific
directly consumed; it should be processed by wet, dry, microorganisms were selected for the starter culture during
or semi-dry methods (Schewan et al., 2012). Wet method the fermentation process of coffee, since it is important
is the most commonly used for coffee processing and it stage to improve the quality of fermentation process and
needs a relatively faster method than dry and semi-dry also the sensory quality of coffee liquour (Massawe and
methods. In the wet method, after harvesting process of Lifa, 2010).
the coffee fruits, the pulp is mechanically removed and
then spontaneously fermented to remove the mucilage Coffee pulp has a high fiber that consisted of cellulose
layer. This method liberated about 40% of solid waste from (49%), hemicellulose (24.5%) and (7.63%) of lignin (Diniyah
coffee pulp (Saenger et al., 2001). The coffee pulp can be et al., 2013). Isolates of Actinomycetes (e.g Streptomyces spp.)
recycled include activities such as composting, feeding to has been proved can utilize the polysaccharides (e.g. starch,
animals, and production of organic fertilizer and biogas cellulose, hemicelluloses) as nutrients for its metabolism
(Rojas et al., 2003). because it can produce extracellular hydrolytic enzymes
(Kokulya et al., 2002). Based on Tuncer et al. (2004),
Spontaneous fermentation process carried out by complex the Streptomyces sp. F2621 produces lignin peroxidase,
microorganisms such as yeast, bacteria, and fungi (Silva endoglucanase, and xylanase enzymes which can degrade
et al., 2008). However, spontaneous fermentation of lignocelluloses. Astuti (2011) stated that S. exfoliatus 42 has
*Corresponding author:
T.C. Sunarti. Department of Agroindustrial Technology, Bogor Agricultural University, Bogor, 16680, Indonesia. Mobile: +6281381055432,
E-mail: [email protected]

Received: 03 May 2015;   Revised: 14 January 2016;   Accepted: 17 January 2016;   Published Online: 11 February 2016

188 Emir. J. Food Agric  ●  Vol 28  ●  Issue 3  ●  2016

 Kurniawati, et al.: Actinomycetes starter on coffee fruits

endoglucanase and exoglucanase activities, while S. exfoliatus with uniform quality. Coffee fruits were washed and rinsed
42 also known having xylanolytic activity (Apriyani, 2012); by using potable water and then poured into fermentation
and Nur (2008) reported that S. costaricanus 45I-3 also chamber, contained 500 g of coffee fruits, and followed by
had xylanolytic activity. In order to obtain the synergistic sterilization by using UV light exposures for 60 minutes.
activity of each strain in this research, the consortium of
S. exfoliatus 42 and S. costaricanus 45I-3 concortia starter Cultivation process
were used to accelerate fermentation process of the whole In the control treatment (spontaneous fermentation),
coffee fruits. coffee fruits that have been sterilized further added with
50 mL of sterile water. In the sample treatment, coffee
There are many reports of the production of enzyme, fruits were added with 50 mL or 10% (v/w) of microbial
antibiotics, organic acids, and bioactive compound which concortia of Actinomycetes as a starter culture, then
conducted by utilizing agricultural waste as substrates incubated for 9 days at 27 °C. Every 3 days the samples
through solid-state fermentation (SSF) (Martins et al.,2013; was observed, and the samples were conducted on two
Murty and Naidu 2011; Prata and Oliviera, 2007; replicates.
Shankaranand, V.S., and Lonsane, B.K. 2003). However,
relatively fewer studies have been conducted on bioactive Concortia of actinomycetes cultivation performance
compound extraction by the action of Actinomycetes The influence concortia of Actinomycetes as starter
through SSF. In the present study, utilization of S. exfoliatus culture addition in SSF to degrade of coffee pulp was
42 dan S. costaricanus 45I-3 starter consortia in the coffee characterized by monitoring the changes in the fiber
fruits fermentation to enhance the yield and quality of component, reducing sugar and total sugar, and active
coffee pulp polyphenol extracts. compounds. After being cultivated, coffee fruits were
further peeled to separate their coffee pulp from the coffee
beans. Coffee pulp were then dried in an oven at 50 °C for
MATERIALS AND METHODS
48 hours. Coffee pulp that has been dried and then ground
Microorganisms and cultures preparation to 40 mesh and used for analyses fiber component (Van
S. exfoliatus 42 and S. costaricanus 45I-3 used in this research Soest et al., 1963) and extracted the active compounds.
were collected from Animal Biotechnology and Biomedical The extraction process was conducted by maceration.
Laboratory, Center for Life Science & Biotechnology, Bogor A mass 25 g of coffee pulp powder was extracted by
Agricultural University. The cultures were re-cultured on 250 mL ethanol: water (80 : 20) solvent under agitation at
a Yeast Starch Agar (YSA) slant which contained: 1.0 g 100 rpm for 24 hour in Erlenmeyer flasks. The extract was
soluble starch, 0.02 g MgSO4.7H2O, 0.05 g K2HPO4, 2.2 g concentrated using a rotary evaporator. Furthermore, the
agar (per 100 mL). For 100 mL of propagation medium, extract was analyzed for sugars and bioactive compounds.
80 mL of YSA media were diluted and supplemented by
1 g of dried coffee pulp powder, then incubated for 7 days Analysis of sugars content which obtained such as total
at 27 °C. sugars were measured by phenol-H2SO4 method (Dubois
et al., 1959), and reducing sugars by DNS method (Miller,
Preparation of starter culture and fermentation 1959). Aliquots (1 mL) and added 2 mL of DNS reagent.
substrates The reaction mixture was incubated for 15 min at 100 oC
The enzyme activity in the coffee pulp media was measured in a water bath. Absorbance was measured at 550 nm by
to determine the growth of microbial concortia of UV-Vis spectrophotometer.
Actinomycetes as a starter culture that is used in the next
cultivation process. Two cockborers of S. exfoliatus 42 Analysis bioactive compounds in coffee pulp such total
and S. costaricanus 45I-3 for each culture were inoculated polyphenol content was measured using a modified Folin-
in propagation medium and incubated at 27 °C by using Ciocalteau method (Singleton and Rossi, 1965), and gallic
a shaking incubator with an agitation speed of 100 rpm acid was used as a standard, and the (%T) by UV-Vis
for 10 days. The supernatant were collected every day for spectrophotometer at a wavelength of 700 nm. The yield
enzyme activity assay. The starter with highest enzyme 1 gr of extracts were mixed with 5 mL Folin-Ciocalteau
activity was used in next cultivation process. Enzyme reagent in 100 mL volumetric flask that contained 50 mL
activity was measured by using DNS (Dinitrosalisilic acid) deionised water. Sodium carbonate solution (15 mL of 20%
method by Miller (1959) with glucose to cellulase activity mv-1 anhydrous sodium carbonate in deionised water) was
and xylose to xylanase activity as the standard. added after 1 minute. The volumetric flask were than made
up to volume with deionised water and after standing at
Preparation of fermentation substrates were conducted by 2 hour at room temperature the absorbance was measured
using whole coffee fruits of Robusta (Coffea canephora) type by UV-Vis spectrophotometer.
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 Kurniawati, et al.: Actinomycetes starter on coffee fruits

Determination of tannin content with buthanol-HCl

method by IAEA (1999) with ethanol-HCl solution
(95%). Solution of ferric (25 ferric ammonium in 2 N
HCl) was prepared by mixing 16.6 mL of HCl in 100 mL
in deionised water to make 2N HCl, and then 2 g of
ferric was dissolved in HCl solution. Mix to analyze
tannins prepared by mixing 0.5 ml of the sample, add
3 mL of butanol-HCl, was added 0.1 mL of reagent Fe
into the tube and shaken using a vortex. tubes containing
material is heated at 90 °C for 60 minutes in water
bath. Absorbance was measured at 550 nm by UV-Vis
spectrophotometer.

Determine catechins were analyzed by High Performance

Liquid Chromatography (HPLC) and anthocyanins Fig 1. The enzyme activities of Actinomycetes concortia in the coffee
pulp medium 1% and incubated at 27°C
measured according to Igelias et al. (2008).

Statistical analysis or exponential phase. The results also showed that

All data were obtained in this research was analysis using Actinomyctes concortia produced higher xylanolytic
statistical analysis. The parameters were observed as fiber activity compared to cellulolytic one, even cellulose is
compounds (cellulose, hemicellulose, lignin, and extractive the main component of coffee pulp.
compound), sugar (total sugar and reducing sugar), and
bioactive compounds (total polyphenol, anthocyanin, Those results are in accordance with the research reported
catechin, and tannin). The data obtained are shown as by Tuncer et al. (2004) who stated that the xylanase
the mean ± standard deviation of 2 replicates and duplo, and cellulase enzymes of Streptomyces sp. FP2621 were
then analyzed using a Completely Randomized Design produced in the growth phase (exponential). The enzyme
(CRD) using SAS software (Statistical Analytic Software activity on Actinomycetes concortia is lower than one
version  9.1). Duncan test α: 0.05 to identify significant isolate of Actinomycetes in CMC and synthetic xylan
differences. media. It is due to nutrients competition in utilizing
substrate for microbial growth. Interactions between
species are not only of synergism or commensalism, but
RESULTS AND DISCUSSION also can be a competition and inhibition (Kato et al.,
2005). Differences in enzyme activity is also caused by
The enzyme activity of actinomycetes concortia
the presence of polyphenols which can be inhibitors to
starter cultur on coffee pulp medium
the enzyme. Polyphenols could bind the enzyme active
The activity of cellulase and xylanase enzymes by site thus inhibiting the activity of the cellulase enzyme
Actinomycetes concortia in coffee pulp medium was (McDaugall et al., 2005; Jurgonski et al., 2013). These
measured to determine the time for preparation of polyphenols were water-soluble compounds generated due
Actinomycetes concortia as a starter that was used in to the reduction of coffee pulp size to 40 mesh as enzyme
the cultivation process (Fig. 1). The concortia enzymes production medium. Differences in enzyme activity is
activities were slower produced compared to single also due caused by the type and substrate concentration
Actinomycetes (Data not shown). The increase of differences. The enzyme has a high specificity to the
enzyme activity was not significant in 1-2 days incubation, substrate. The use of different medium causes differences
since it was that microbial concortia of Actinomycetes in resulted enzyme activity (White, 1995).
in the adaptation phase (lag phase). It is also due to
the microbes has not hydrolyze cellulose and xylan as Cultivation performance from actinomycetes concortia
carbon source but using available reducing sugar in coffee Cultivation of Actinomycetes concortia in the coffee fruits
pulp. Fontes et al. (2000) reported that the growth of causing the change in fiber contents of coffee pulp. The
xylanase producing microbes was tested using glucose change was associated with the presence of microbes
and xylan as the carbon sources resulted in more rapid and their ability to secrete extracellular enzymes. In the
cell growth on glucose medium compared with cell control treatment (B), reduction of fiber content was
growth on xylan medium. The high activity produced smaller than sample treatment (A) (Table 1). According
from 6 days fermentation, indicating that the growth of to Dalzell et al. (1997) decomposition of organic material
Actinomycetes concortia reached the maximum point by microbial and water needed oxygen and nutrients from
190 Emir. J. Food Agric  ●  Vol 28  ●  Issue 3  ●  2016
 Kurniawati, et al.: Actinomycetes starter on coffee fruits

organic matter as a source of energy and then release CO, There were differences in the decrease of cellulose,
water and heat energy, causing the weight of the material hemicellulose and lignin levels in coffee pulp during
decreases. A decrease in weight due to the release of CO2 fermentation using Actinomycetes concortia. The highest
and other compounds. Those results are in accordance with decrease percentage in the lignin and cellulose levels in
the research reported by Tuomela et al., (2000) organic coffee pulp was on day-6 at 4.19% of lignin, 29.11% of
material of lignocellulosic utilized as a carbon source for cellulose, and on day-9 at 16.89% of hemicellulose (Fig. 2).
microbial metabolic processes that produce energyand The decrease in lignin content in coffee pulp substrate
release CO2 and the end product is more simple line with occured due to lignin peroxidase enzyme activity produced
produced extracellular enzymes. Weight loss also showed by S. exfoliatus 42. Apriyani (2013) stated that S. exfoliatus
that the consortium actinomycetes able to describe the 42 was capable of degrading lignin in corn cobs, causing a
components used in fiber include cellulose, hemicellulose, 4.8% decrease in lignin content. Peroxidase enzymes could
and lignin in coffee pulp. degrade the substrate such as phenols, aromatic amines and
several components such as alkyl peroxide (Jing Li et al.,
The decrease of fiber content in control (spontaneous 2009). The decrease in lignin content occurred after 3 days
fermentation) did not occur significantly, due to the growth of incubation, and the highest decrease in lignin content was
of indigenous bacteria on the coffee fruits. According to occured on 6th day. Tuncer et al. (2004) stated that Streptomyces
Silva et al. (2008), naturally, the coffee processing through the sp. FP2621 has lignocellulolytic activity as peroxidase.
spontaneous fermentation involved microorganisms such as Peroxidase activity increased with cell biomass growth and
Bacillus sp. which was able to hydrolyze cellulose due to its achieved optimum activity after 4 days of incubation.
ability to produce cellulase (Coughlan and Mayer, 1991). The
role of Actinomycetes starter in the cultivation process was The highest decrease in cellulose content occured on day-6.
related to its ability to produce extracellular enzymes, such It was linked with the production of cellulase enzyme by
as cellulase, xylanase and lignin peroxidase (Apriyani, 2012). S. exfolatus 42. The result of this degradation is faster than
Microbes have the capabilities to produce the enzymes for the research conducted by Tuncer et al. (2004). The highest
assimilate organic matter by degrade components of the
substrate; more complex substrates is used, more complex
enzyme is required to degrade the substrate (Tuomela et al.,
2000). Soto et al. (2008) reported the decreasing in cellulose
and hemicellulose fibers in Borage officinalis was due to the
use of cellulase and xylanase enzymes. The presence of
enzyme that was produced by Actinomycetes concortia was
effectively used to degrade complex organic components
into a simple molecule called monosaccharide which can be
used as a carbon source for microbial cells.

The ability of an enzyme to degrade the fiber components

depends on the activity of the enzyme possessed. Greater
enzyme activity means higher enzyme ability to degrade
lignocellulose components. Kammerer et al. (2005) stated that
the use of enzyme in the polyphenols extraction from grape
processing waste must have the relevant processes such as the Fig 2. The decrease of lignin, cellulose, and hemicellulose after
type of enzyme, the enzyme-substrate ratio and temperature. incubation at 27°C

Table 1: The composition of the substrate component after cultivation at 27 °C

Fiber compound Treatment Incubation time (day)
(%) 0 3 6 9
Cellulose A 56.30±0.2a 49.13±0.6b 30.44±0.7c 28.65±0.4c
B 56.38±1.2d 51.57±0.3d 48.33±0.1e 45.24±1.3f
Hemicellluose A 23.65±0.3 a
18.29±0.4 a
16.00±0.5c 11.14±0.3d
B 23.31±0.2 a
22.92±0.7 b
15.68±0.7c 10.28±0.4d
Lignin A 6.10±0.1 a
3.40±0.4 b
2.90±0.2b 2.83±0.1b
B 6.10±0.1a 5.40±0.5c 4.90±0.4c 4.89±0.5c
Extractive compound A 13.95±0.1a 29.18±0.2b 50.66±0.3c 57.38±0.1c
B 14.21±0.1d 20.11±0.1d 30.09±0.3e 36.59±0.1f
Data: Mean±standard deviation; α=0.05, Note: A: Sample (Actinomycetes consortium), B: Control (Spontaneous fermented)

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 Kurniawati, et al.: Actinomycetes starter on coffee fruits

hydrolysis of straw substrate with endoglucanase produced occoured on 3th day of incubation, but a few decreased
by Streptomyces sp. FP2621 was on 7th day (Tuncer et al., on 6 th day of incubation. It is because on 6 th day,
2004). Jager et al. (2010) stated that the microbes could Actinomycetes concortia only converted oligosaccharides
hydrolyze crystalline cellulose through cellulase enzymes into monosaccharides. The yield of reducing sugar in
depends on the crystallinity of cellulose. Cellulase enzyme control was a few increased or constant, in contrast to the
could hydrolyze cellulose in plant cell walls. This hydrolyze sample treatment. It is due to microbes that are present
process caused insoluble fiber content decrease into more during the fermentation process. In the sample treatment,
soluble and simple components (Yoon et al., 2005). Wang reducing sugar increased on 3th day of incubation and
et al. (2008) describe that the cellulase enzymes produced reached the highest on 6th day of incubation. It is in
by microbes to utilize cellulose involve the combined accordance with the highest degradation of complex fiber
hydrolysis enzymes, consisting of endoglucanase, component (Fig. 2). Reducing sugar is a product of cellulose
exoglucanase (cellobiohydrolase), and ß-glucosidase. and xylan degradation by enzyme generated microbes.
Degradations of xylan and cellulose produce monomeric
The highest decrease in hemicellulose content in 9th day sugars in the form of reducing sugar. According to Saha
of incubation was related to xylan hydrolyzation by (2004), hydrolysis of cellulose produces glucose monomers
xylanase enzyme, because xylan is the largest component and cellobiose oligomers, while xylan produces xylose,
of hemicellulose. The xylanolytic enzyme system that arabinose, and xylooligosaccharide. According to Yoon
carries out the xylan hydrolysis is normally composed of et al., (2005) fiber hydrolysis using cellulase enzymes obtained
a repertoire of hydrolytic enzymes, including endo1.4-ß- some sugar components are monosaccharides (glucose,
xylanase that cleaving the glycosidic bonds and in liberating fructose, galactose, and arabinose), cellooligosaccharides
short xylooligosaccharides, 1.4 ß-D-xylosidase to convert (cellopentaose, cellotetraose, cellotriose, and cellobiose), and
xylooligosaccharide into xylose. The side constituent groups galactooligosaccharides (galactotetraose and galactotriose).
of xylan will be released by α-L-arabinofuranosidase, α-D-
glucuronidase, and acetyl xylan esterase to arabinose, The degradation of lignocellulose component would
glucuronic acid, and acetate (Subraminayan and Prema, affect the extraction yield of bioactive compounds in
2002). The hydrolysis mechanism of xylan is initiated by coffee pulp. The result of total polyphenols extract
α-arabinofuranosidase that capable of hydrolyze xylan into obtained was higher in the process using starter
L-arabinose and xylobiose. The cleavage of branch chain culture of Actinomycetes concortia compared with the
of xylan will facilitate the hydrolysis of xylanase by exo- spontaneos cultivation (Table 3). The statistical analysis
xylan and β-xylosidase. The main products of hydrolysis using α: 0.05 also showed that the use of Actinomycetes
of these enzymes were xylose, arabinose, xylotetrose, and concortia was significant impact on the yield of bioactive
xylotriose (Puspaningsih, 2004). compounds extract and Duncan’s test also showed
significant differences between control and sample
Lignocellulose hydrolysis by lignocellulolytic enzymes treatment. In the control, there were limited number
also affects to the yield of the extracts such as total of microorganisms during fermentation that caused
sugar, reducing sugar, and bioactive compounds. The slightly increased in polyphenol extracts obtained. In
determination of components is done by extracting using fermentation, the presence of hydrolytic enzymes not
a polar solvent is ethanol. Therefore, in this study the only causes the degradation of the cell wall, but also
measured components of the extraction of the bioactive might affect phenolic compounds stability. Hydrolysis
components that are polar. Polyphenols are a group of using an enzyme decreases the viscosity of the substrate,
polar compounds which contains an OH group (Shi et al. reduce the attractive forces between molecules and
2003). Measurement of the sugar component is the total decrease the stability of the interaction between uronic
sugar and reducing sugar, whereas the bioactive compounds acids, proteins, and tannins. In contrast, the breaking of
include total polyphenols, anthocyanins, tannins and a cell wall polymer increases permeability and porosity of
catechins. The result of the extraction coffee pulp showed the cell, enchance solubility of internal cell components
that the total sugar content was increasing after treated by with a consequent increase in the concentration of
Actinomycetes concortia compared to the spontaneous phenolic compounds and the antioxidant activity of
cultivation. The analysis result showed that the use of the extract (Cerda et al., 2013). Therefore, the addition
Actinomycetes concortia affected the yield of sugar content of Actinomycetes concortia culture can improve the
and Duncan’s test also showed significant differences yield of bioactive compounds particularly polyphenol
between control and sample treatment (Table 2). compound extracts. The yield analysis of secondary
metabolites include four of bioactive compounds
The highest of total sugar content from the cultivation of coffee pulp, such as total polyphenols, tannins,
process using starter culture of Actinomycetes concortia anthocyanins, and catechins. Ramirez-Coronel et al. (2004)
192 Emir. J. Food Agric  ●  Vol 28  ●  Issue 3  ●  2016
 Kurniawati, et al.: Actinomycetes starter on coffee fruits

Table 2: Yield sugar was extracted after cultivation at 27 °C

Compound Treatment Incubation time (day)
0 3 6 9
Total sugar (mg mL−1) A 11.94±0.62c 19.60±1,62a 16.25±0.85b 10.82±0.87c
B 12.17±1.31c 11.98±1.31c 11.89±1.30c 12.35±0.00c
Reducing sugar (mg mL−1) A 2.06±0.00b 4.82±0.01 a
4.86±0.01a 1.73±1.01c
B 2.06±0.16b 2.53±0.18 b
2.78±0.18b 2.67±0.19b
Data: Mean±standard deviation; α=0.05, Note: A: Sample (Actinomyset concorcia), B: Control (Spontaneous fermented)

Table 3: The yield of the bioactive compounds after cultivation at 27°C

Compound Treatment Incubation time (day)
0 3 6 9
Polyphenol (mg mL−1) A 0.75±0.01d 0.76±0.00d 1.20±0.00a 1.15±0.04b
B 0.75±0.03d 0.75±0.03d 0.86±0.02c 0.87±0.02c
Tannin (%) A 2.49±0.15d 2.77±0.17c 4.20±0.14a 2.94±0.00c
B 2.28±0.12d 2.70±0.12c 4.34±0.08a 3.97±0.02b
Anthocyanin (mg g )−1
A 57.68±0.23d 58.00±0.2 c
109.95±0.1a 68.00±0.1c
B 57.68±0.2d 59.32±0.2 c
101.59±0.1a 66.36±0.1b
Catechin (mg mL )
−1
A 0.24 10.38 0.79 0.34
B 0.24 6.99 0.61 0.32
Data: Mean±standard deviation; α= 0.05, Note: A: Sample (Actinomyset concorcia), B: Control (Spontaneous fermented)

found the bioactive compounds consisted of four main obtained in the sampel treatment was lower compared with
classes of polyphenols in coffee pulp, i.e.  flavan-3-ols, the control treatment (Table 3). The same study results
hydroxycinnamic acids, flavonols, and anthocyanidins. that use Streptomyces sp. through solid state fermentation
showed the decrease in polyphenols content in coffee
The yield of extraction showed that the highest total pulp (Orozco et al., 2008). The different results shown by
polyphenol was obtained on 6 th day of incubation. Moreno-Peres et al. (2010), in which tannin extract from
According to Huang et al. (2007), a mixture of Aspergillus grapes increased after fermented using pectinase and
oryzae and Trichoderma reesei that produces cellulase and β-galactosidase enzymes. The reduction of polyphenols
xylanase enzymes can improve the yield of ellagic acid content, particularly tannin from fermented coffee pulp,
extract which is classified as polyphenols. Laroze et al. can be utilized as animal feed. It caused by tannins can
(2010) and Collao et al. (2007) also reported that the use inhibit the growth of fiber-degrading bacteria in the
of commercial enzyme mixture such as cellulase and digestive tract of ruminants (Ozkose et al., 2011). Increased
hemicellulase can enhance the yield of polyphenol extracts tannin extract in both control and sample treatments was
that can be used as antioxidant from Raspberry and Oenothera followed by an increase in its monomer called catechin.
biennis. Maier et al. (2008) also stated that the use of a The yield of catechins increased dramatically after 3 days
mixture of two types of commercial enzymes can improve of incubation.
the yield of polyphenol extracts in grapes. Extraction of
phenolic antioxidants from vegetables using enzymes may The increase of total polyphenols was apparently due to
occur through hydrolytic degradation of polysaccharides increase one of its components, such as anthocyanin. In
in cell wall. These phenolics are bound to lignin and the sample treatment, the yield of anthocyanin extract
polysaccharides by hydrogen or hydrophobic bonds. In was higher than control. Polyphenolic compounds such
addition, other mechanisms may also be carried out by as anthocyanins have covalent bonds to the cell wall.
enzymes which are directly cleave the ether or ester bonds Those results are in accordance with the research reported
between the phenols and plant cell wall polymers (Pinelo by Ramirez-coronel et al., (2004) who stated that the
et al., 2008). Increase of total polyphenolic compounds increased of anthocyanin extracts yield was occurred
also occurs due to the release of the bond between the after 3 days of incubation. According to Jurgonski et al.
components of cellulose, hemicellulose and lignin with (2013), β-glucosidase enzyme can increase anthocyanin
polyphenolic compounds. Polyphenolic compounds are extracts from L. caerulea. Extraction of polyphenol and
covalently bound to the cell wall. According to Gonzales anthocyanin from Blackcurrent fruits can improved the
et al. (2011), solid fermentation using Aspergillus tamarii was yields by cellulase and hemicellulase enzymes derived
able to improve the yield of phenolic compound extracts in from Trichoderma spp. (Kapasakalidis et al., 2009). Prata
the form of hydroxycinnamic acid from the coffee pulp that and Oliveira (2007) described the use of fresh coffee pulp
can be used as an antioxidant compound. Tannin extract as a potential source of natural dye due to the content of
Emir. J. Food Agric  ●  Vol 28  ●  Issue 3  ●  2016 193
 Kurniawati, et al.: Actinomycetes starter on coffee fruits

cyanidin-3-rutinoside anthocyanin. Murthy et al. (2012) on oil extraction and antioxidant recuperation from Oenothera
biennis by cold pressing. Grasas y Aceites. 58: 10-14.
also reported that the red color of coffee pulp contained
cyanidin-3-rutinoside and cyanidin-3-glucoside that Coughlan, M. P. and F. Mayer. 1991. The cellulose-decomposing
bacteria and their enzymes systems. Prokaryotes. 1: 460-516.
potential as antioxidants and natural dye foods. In addition,
Dalzel, H. W., A. J. Biddlestone, K. R. Gray and K. Thurairajan. 1997.
Pinelo et al. (2008) stated that β-galactosidase enzyme from Soil management: compost production and use in tropical and
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IAEA. 1999. Working Document. Quantification Taninns iin Tree
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Titi Candra Sunarti through Institute of Research and colour development, anthocyanin content, fruit quality, and
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