The Effect of Mixed Biological Pretreatment and PEG 4000 on Reducing Sugar Production from Coffee Pulp Waste
Corresponding email: triw@chem-eng.its.ac.id
Published at : 24 May 2019
Volume : IJtech
Vol 10, No 3 (2019)
DOI : https://doi.org/10.14716/ijtech.v10i3.2900
Iswanto, T., Hendrianie, N., Shovitri, M., Altway, A., Widjaja, T., 2019. The Effect of Mixed Biological Pretreatment and PEG 4000 on Reducing Sugar Production from Coffee Pulp Waste. International Journal of Technology. Volume 10(3), pp. 453-462
| Toto Iswanto | Department of Chemical Engineering, Institut Teknologi Sepuluh Nopember, Sukolilo, Surabaya 60111 Indonesia |
| Nuniek Hendrianie | Department of Chemical Engineering, Institut Teknologi Sepuluh Nopember, Sukolilo, Surabaya 60111 Indonesia |
| Maya Shovitri | Department of Biology, Institut Teknologi Sepuluh Nopember, Sukolilo, Surabaya 60111, Indonesia |
| Ali Altway | Department of Chemical Engineering, Institut Teknologi Sepuluh Nopember, Sukolilo, Surabaya 60111 Indonesia |
| Tri Widjaja | Department of Chemical Engineering, Institut Teknologi Sepuluh Nopember, Sukolilo, Surabaya 60111 Indonesia |
Biological methods using bacteria and fungi
are regarded as more economically viable and environmentally friendly
alternatives for improving lignocellulosic degradation. Coffee pulp waste (CPW)
as a lignocellulosic biomass is abundant and has potential as a reducing sugar
feedstock. However, it contains lignin as a matrix polymer, which associated
with pectin and cover the cellulosic microfibrils and make it difficult to be
digested during the bioprocess. In this study, the performance of biological
pretreatment in reducing lignin and pectin using a co-culture of Bacillus subtilis (BS), Aspergillus niger (AN), or Trichoderma reesei (TR) has been
investigated. The pretreatment of the CPW was made using various microbial
ratios in an aerobic stirred-bioreactor and incubated at 30o C, pH 5
for 7 days. Removal of lignin and pectin was analyzed during the pretreatment
process. PEG 4000 as a surfactant was
used and its effect on the yield of
reducing sugar production from pretreated CPW using a A. niger and T. viride
(TV) co-culture with a surfactant to substrate
ratio of 1:1 (w/w) was investigated. A culture without surfactant was used as a
control. The results reveal that the best lignin and pectin removal was 99.9%,
when using a co-culture of AN and TR with a ratio of 1:1 (v/v) and of BS and TR
with a ratio of 2:1 (v/v). The cellulose content of CPW in these co-cultures
was 86.99% (w/w) and 81.61% (w/w), respectively, and the reducing sugar
concentration obtained was 12.5 g/L and 9.74 g/L respectively. In further
hydrolysis of pretreated CPW using a AN:TV (2:1) co-culture with the addition
of surfactant, the yield of reducing sugar obtained was higher than that of the
control, at 20.69%. Use of PEG 4000 as a surfactant had a positive effect on
enhancing the yield of reducing sugar from coffee pulp waste.
Biological pretreatment; Coffee pulp waste; Hydrolysis; Reducing sugar; Surfactant
The utilization of agro-industrial residues
as lignocellulosic biomass to produce
more valuable material has encouraged the improvement of advanced
biotechnological innovations, principally in enzyme and fermentation
technology, which have ultimately had an impact on the reduction of
environmental pollution problems (Pandey et al., 2000). One of the agro-industrial residues that is available in abundance in
Indonesia, but which has particular problems associated with its utilization due
to the presence of anti-nutritional factors
such as caffeine,
In the
biotechnological approach, CPW can be converted into reducing sugar as a
substrate to produce biofuels such as ethanol and biogas. To obtain the optimum
yield of reducing sugar, a pretreatment
process is needed to remove the unnecessary matrix polymers such as lignin and
pectin surrounding the cellulosic microfibrils. Pectin is a heterogeneous
polysaccharide, whose molecular structure, weight and functional properties
depend on its agro-waste source (Kusrini et al., 2018). Pectin has the function
of cross-linking cellulose and hemicellulose fibers, providing rigidity to the
cell wall (Abbott & Boraston,
2008). This is in line with Menon and Rao (2012), who state that hemicelluloses, amorphous
polymers of different sugars, and other polymers such as pectin, attach the
microfibrils of cellulose, which are stabilized by hydrogen bonds and covered
by lignin, making the biomass difficult to be digested in the bioprocess.
Physicochemical pretreatment methods have been widely developed, such
as steam explosion (Guo et al., 2011), or use of dilute acid (Dussán et al.,
2014), alkali (Menezes et al., 2014) or oxidation, or varied combinations of
these, but such processes usually involve high temperature, pressure and cost (Kumar et al., 2009). Since chemical or physical methods are
not able to provide selective removal, are expensive, and are not
environmentally friendly, microbial and enzymatic techniques for lignin and
pectin removal could be alternative advantageous methods.
The co-culture
strategy has been assessed as a more efficient way to treat lignocellulosic
waste (Menon & Rao, 2012). The combination of cultures can consist
of two, three, or a consortium of bacteria or fungi or a mixture of them, which
can be applied in many biological production processes. In the case of lignin and pectin removal, previous
studies have reported that A. niger
and T. reesei are capable of
degrading lignin (Adav & Sze, 2014;
Asses et al., 2018). Moreover, Roussos et al. (1995) found that some
strains of A. niger are capable of
degrading caffeine. B. subtilis can degrade lignin and pectin (Gummadi & Kumar, 2005; Torimiro & Okonji, 2013; Cragg et
al. 2015). Juliastuti et al. (2018) successfully achieved the combination of
three strains, A. niger, Pseudomonas putida, and T. harzianum to treat coffee pulp in
degrading lignin and anti-nutritional factor for a biogas feedstock.
Besides applying
the pretreatment, the use of surfactants (i.e. PEG 4000) in the hydrolysis of
lignocellulosic substrates has produced good effects. Surfactants, as
surface-active additives, affect the interaction between enzymes such as
cellulase and substrates (Li et al., 2012), leading to an increase in the total
amount of reducing sugar after the hydrolysis process. PEG is presented in
varying molecular weights (e.g. PEG 4000, 6000 and 8000); at the lowest
molecular weight (PEG 4000) the amounts of reducing sugar produced were 7% and
13% higher than those from PEG 6000 and 8000 respectively (Iveti? et al., 2014). However,
the effects of the addition of the same surfactants on the obtained hydrolysis
product may be different, depending on
the operating conditions, initial chemical content in the substrate, and
treatment method during the hydrolysis
process.
Therefore,
this study conducts an investigation to determine the best co-culture ratio
involving A. niger, T. reesei and B. subtilis to develop a co-culture for degrading lignin and pectin
at a faster degradation rate and which is able to tolerate the presence of the
anti-nutritional factors that effect their growth. The obtained CPW from the best
pretreatment condition was further hydrolyzed
using a co-culture of A. niger and T. viride with the addition of PEG 4000. The effects of the
co-culture on the reducing sugar obtained from the hydrolysis process were also
investigated and described.
The
microbial method in the form of a fungal and bacterial mixture has produced
satisfying results in the degradation of lignin and pectin in coffee pulp
waste, with more than 99% removal of those compounds. Due to the high cellulose
concentration (more than 80%) in pretreated coffee pulp waste, two co-cultures,
AN:TR (1:1) and BS:TR (2:1), were chosen for
the following hydrolysis process. Subsequently, the hydrolysis of pretreated
CPW using A. niger and T. viride with the addition of
surfactant resulted a higher yield of reducing sugar than that without
surfactant. The highest yield of reducing sugar resulted from the pretreated
CPW by AN:TR (1:1) and hydrolyzed by the
AN:TV (2:1) co-culture with the addition
of PEG 4000, at 20.69%. In this co-culture, PEG 4000 as surfactant made a
difference of 3.4% in reducing sugar, compared to without surfactant.
Hopefully, the co-culture ratio obtained can indicate a direction for further
study, focusing on the kinetics of its cell growth, which is important for
developing an economical bioprocess of reducing sugar production.
The authors thank Atikah Badriya Husein and Dwi Ayu Primaningrum who
have provided coffee pulp waste and selfless help. We also thank all members of Biochemical Technology Laboratory
and Wastewater Treatment Laboratory, Institut Teknologi
Sepuluh Nopember for their endless support.
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