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Waste Management 49 (2016) 320325

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Waste Management
journal homepage: www.elsevier.com/locate/wasman

Integrated bioethanol and biomanure production from potato waste


Anjani Devi Chintagunta a, Samuel Jacob b, Rintu Banerjee b,
a
b

Advanced Technology Development Centre, Indian Institute of Technology, Kharagpur 721302, West Bengal, India
Agricultural & Food Engineering Department, Indian Institute of Technology, Kharagpur 721302, West Bengal, India

a r t i c l e

i n f o

Article history:
Received 1 May 2015
Revised 23 July 2015
Accepted 10 August 2015
Available online 24 August 2015
Keywords:
Bioethanol
Biomanure
Potato waste
Solid state fermentation

a b s t r a c t
Disposal of potato processing waste and the problem of pollution associated with it is a vital issue that is
being faced by the potato processing plants. The conventional peeling methods presently followed in the
processing plants for removing the potato peel, also result in the loss of some portion of the mash which
is rich in starch. Indiscriminate discharge of the waste causes detrimental effects in the environment, so
this problem can be resolved by successful utilization of the waste for the generation of value added
products. Hence, the present work focuses on integrated production of bioethanol and biomanure to utilize the waste completely leading to zero waste generation. The first part of the work describes a comparative study of ethanol production from potato peel and mash wastes by employing co-culture of
Aspergillus niger and Saccharomyces cerevisiae at various incubation time (24120 h) instead of application
of enzymes. The solid state fermentation of potato peel and mash inoculated with co-culture, resulted in
bioethanol production of 6.18% (v/v) and 9.30% (v/v) respectively. In the second part of the work, the residue obtained after ethanol production was inoculated with seven different microorganisms (Nostoc muscorum, Fischerella muscicola, Anabaena variabilis, Aulosira fertilissima, Cylindrospermum muscicola,
Azospirillium lipoferum, Azotobacter chroococcum) and mixture of all the organisms in equal ratio for nitrogen (N), phosphorous (P) and potassium (K) enrichment. Among them, A. variabilis was found to enrich N,
P and K content of the residue by nearly 7.66, 21.66 and 15 fold than that of the initial content, ultimately
leading to improved N:P:K ratio of approximately 2:1:1. The application of simultaneous saccharification
and fermentation (SSF) for the conversion of potato waste to ethanol and enrichment of residue obtained
after ethanol production with microorganisms to be used as manure envisages environmental
sustainability.
2016 Published by Elsevier Ltd.

1. Introduction
Changing life style and craving for processed foods are factors
responsible for the significant growth of global food processing
industries. Among several varieties of processed foods, the products from potatoes are in great demand. Potato is a starchy tuber
with carbohydrates constituting as much as of 1330% with a little
protein (0.74.6%) (Puttongsiri et al., 2012). The production of
potato in India for the year 201213 was reported to be 45.3 million tonnes from 1.99 million hectares of land with an yield of
22,763.1 kg/ha (Agricultural statistics at a glance, 2013). The
potato processing industry has multiplied in India owing massive
potato production and a booming demand for its products in the
market. Potato peeling is the initial step in any potato processing
industry and losses caused during this stage may range from 15%
to 40% according to the method followed for peeling (Arapoglou
Corresponding author.
E-mail address: rb@iitkgp.ac.in (R. Banerjee).
http://dx.doi.org/10.1016/j.wasman.2015.08.010
0956-053X/ 2016 Published by Elsevier Ltd.

et al., 2009). The processing loss during production must be


reduced by employing efficient peeling methods to obtain maximum economic returns and at the same time the disposal problem
needs to be addressed by converting the waste into value added
products.
Production of consumable alcohol from the food waste can be
an impressive alternative. Growing per capita spirit consumption
and escalating demand for premium brands are motivating the
growth of global spirits manufacturing market. Although the
industry was hit hard by the economic downturn during 2009, it
has picked up slowly with an average growth rate of 2.2% annually
upto 2013. In 2013, global market revenue rose to 9.5% and
expected to attain 9.8% by 2018 (Mynews desk, 2013). KawaRygielska et al. (2012) reported the utilization of by-products generated during potato granule processing as feedstock for ethanol
production. Ethanol production from starchy biomass generally
involves various steps like liquefaction, saccharification and fermentation (Pervez et al., 2014). In the present study, coculture of
A. niger and S. cerevisiae were used to obtain improved ethanol pro-

A.D. Chintagunta et al. / Waste Management 49 (2016) 320325

duction through simultaneous saccharification and fermentation


process. Ado et al. (2009a) reported that the synergistic metabolic
interactions between A. niger and S. cerevisiae in a starch medium
enhances amylolytic activity and total ethanol yield by preventing
accumulation of inhibitory concentrations of reducing sugar.
The solid residue obtained after ethanol production still contains certain amount of nutrients which upon further enrichment
can be effectively utilized as biomanure, an efficient way of controlling the solid waste accumulation in the environment. The
enriched residue is an excellent organic manure containing all
the essential nutrients vital for plant growth and humus to develop
the soil structure (Bhattacharyya and Banerjee, 2007).
The deficiency of nitrogen is usually met by supplementing the
soil with the chemical fertilizers, whose long term application
leads to soil degradation and other environmental issues. The
import of the chemical fertilizers as percentage value of consumption for the year 20122013 was estimated to be Rs.( ) 4,04,120
million (Chemicals and Petrochemicals Statistics at a Glance,
2014). In the present study, an attempt has been made to enrich
the waste by using blue green algae which are capable of fixing
nitrogen, an essential element for plant growth. Ananya et al.
(2014) reported excretion of vitamins, amino acids and hormones
like auxin, gibberellin etc. by algae which involve in promoting
plant growth. The administration of biomanure is not only a cost
effective way of enriching soil but also a better solution for environmental problems caused by the excessive usage of chemical
fertilizers.
Thus, the present work focuses on a comparative study of ethanol production from potato peel and mash wastes through simultaneous saccharification and fermentation employing co-culture
of A. niger and S. cerevisiae followed by treatment of mixed solid
residue with different nutrient enriching microorganisms for its
utilization as biomanure.

2. Materials and methods


2.1. Substrate
Potato peel and mash waste generated during processing of
potato products were collected from the local plants based in
Kharagpur, West Bengal, India.

2.2. Microorganisms
2.2.1. Inoculum for bioethanol production
Aspergillus niger and Saccharomyces cerevisiae cultures were
obtained from Microbial Biotechnology and Downstream Processing Lab, IIT Kharagpur, West Bengal, India. A. niger was maintained
on Potato Dextrose Agar (PDA) medium at pH 5.6 0.2 and 25 C
whereas S. cerevisiae was grown in medium containing glucose
(2%, w/v) and yeast extract (0.5%, w/v) at pH 6.5 and 37 C.

2.2.2. Inoculum for biomanure production


The blue green algae viz., Nostoc muscorum, Fischerella muscicola, Anabaena variabilis, Aulosira fertilissima, Cylindrospermum
muscicola selected for N, P and K enrichment studies were procured
from Vishwabharathi University, Kolkata and cultured in BG11
medium as reported by Rippka et al. (1979) at pH 7.5 and 25
28 C excluding nitrate or ammonium source as they generate
heterocysts in the nitrogen deficient medium (Pankaj, 2008;
Pereira et al., 2005; Saville et al., 1987). Azospirillium lipoferum
and Azotobacter chroococcum were cultured in nutrient broth medium at pH 7.3 0.2 and 3537 C.

321

2.3. Methods
Moisture, protein, reducing sugar, cellulose and starch content
of potato wastes were determined by following standard methods
(AOAC, 1965; Lowry et al., 1951; Miller,1959; Updegraff, 1969;
Hodge and Hofreiter, 1962). The estimation of N, P and K content
in the potato waste was done by Kjeldahl, spectrophotometric
and flame photometric methods (Scan test method, 1986;
Chapmann and Pratt, 1961; Hegedus and Pungor, 1955) respectively. The reducing sugar and ethanol content were estimated
by dinitrosalicylic acid and potassium dichromate method respectively (Miller, 1959; Seo et al., 2009).
2.4. Experimental setup for bioethanol production
Solid state fermentation process was established with two substrates i.e., potato peel and mash and ethanol production from both
the substrates was observed by employing co-cultures. The experimental set up with each substrate consists of 5 treatments each
containing 100 g (wet weight) of substrate mixed with 20 ml of
modified Czapekdox medium. A. niger (1%, v/w) containing
2.5  106 spores/ml was added to all the 5 treatments. S. cerevisiae
(10%, v/w) (Ado et al., 2009b) was inoculated to the first treatment
after 24 h of A. niger addition. To second treatment, S. cerevisiae
(10%, v/w) was inoculated after 48 h so on and fifth one after
120 h respectively and incubated at 37 C (Fig. 1). Inoculum free
substrate was used as control. Small aliquots were drawn from
each treatment at specific time intervals (24168 h) and centrifuged at 2000 rpm for 5 min. The clear supernatant was collected and analyzed for ethanol content.
2.5. Experimental setup for biomanure production
After bioethanol production, the liquid portion containing the
ethanol was separated from the solid content of both the substrates by squeezing through a cheese cloth. The solid residue (peel
and mash) was collected, mixed, characterized and employed for
nutrient (N, P, K) enrichment to convert into biomanure. A set of
eight treatments, each containing 100 g of mixed residue was
arranged and 10% (v/w) inoculum of each organism like N. muscorum, F. muscicola, A. variabilis, A. fertilissima, C. muscicola, A. lipoferum, A. chroococcum was added individually and as a mixture in
equal proportion and incubated at 2527 C. A control was also
maintained without the addition of inoculum. The residue was
checked for NPK enrichment weekly upto 13 weeks. The schematic
representation of integrated production of bioethanol and biomanure was shown in Fig. 1.

3. Results and discussion


3.1. Characteristics of potato peel and mash wastes
The moisture, starch, cellulose, reducing sugar and protein content of potato peel and mash wastes are presented in Table 1. It has
been observed that potato peel and mash wastes have considerable
starch content (28.52% 0.17 and 49.78% 1.2 (w/dry weight)),
cellulose content (5.69% 1.6 and 2.31% 1.2 (w/dry weight)),
meagre fermentable reducing sugar (0.073% 0.0043 and
1.33% 0.10 (w/dry weight)) and protein content (0.082% 0.002
and 0.16% 0.001 (w/dry weight)) respectively. The reducing sugar
content in the potato peel was in accordance with that reported by
Khawla et al. (2014) but varied in starch and protein content. The
probable reason for such variation could be due to the influence of
climatic conditions, methods adopted for potato processing etc.

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Fig. 1. Schematic representation of efficient utilization of solid potato wastes.

Table 1
Characteristics of potato waste.

a
b

Parameter

Units

Moisture
Starch
Cellulose
Reducing sugar
Protein

(%,
(%,
(%,
(%,
(%,

w/w)a
w/w)b
w/w)b
w/w)b
w/w)b

Potato peel

Potato mash

86.5 0.01
28.52 0.17
5.69 1.6
0.073 0.004
0.082 0.002

76.8 0.02
49.78 1.2
2.31 1.2
1.33 0.10
0.16 0.001

Wet weight.
Dry weight.

3.2. Comparison of ethanol production between potato peel and mash


The ethanol production profile of potato peel and mash waste
inoculated with co-culture of A. niger and S. cerevisiae is shown
in Fig. 2. The ethanol production in co-culture inoculated peel
and mash started within 24 h of incubation time. Significant ethanol production of 6.18% (v/v) was obtained in 120 h of incubation
time from the first treatment of peel in which S. cerevisiae was
inoculated after 24 h of A. niger addition. Arapoglou et al. (2010)
reported 7.58 g/L of ethanol production from potato peel waste
by enzymatic saccharification and fermentation using S. cerevisiae.
Itelima et al. (2013) reported ethanol yield of 10.08 (%, v/v) from
corn cobs after 7 days of fermentation with co-culture of A. niger
and S. cerevisiae inoculated simultaneously. The maximum ethanol
production of 9.30% (v/v) was obtained in 72 h of incubation time
from the second treatment of mash.
The ethanol yield and average ethanol productivity rate from
potato peel was found to be 48.76 g/L and 0.406 g/L/h respectively
whereas in mash it was observed to be 73.4 g/L and 1.02 g/L/h
respectively. Approximately, 80.62% and 83.78% of starch hydrolysis was observed in the co-culture inoculated peel and mash
respectively. Significant starch hydrolysis in the co-culture inoculated peel and mash depicts an efficient action of amylase pro-

Fig. 2. Ethanol production profile in co-culture inoculated (a) peel and (b) mash.

duced by the A. niger for effective conversion of starch to


reducing sugar which was thereby used up by the yeast for ethanol
production. It was reported that solid state fermentation holds
tremendous potentials for the production of amylase by A. niger
(Suganthi et al., 2011) which have contributed to the effective
starch hydrolysis in co-culture inoculated substrate.

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A.D. Chintagunta et al. / Waste Management 49 (2016) 320325

3.3. Nutrient enrichment of residual potato wastes


The residual potato wastes after bioethanol production was collected, mixed and analyzed for initial N, P and K content (Table 2).
The potato peel and mash wastes were mixed to enhance the nutrient content of the residue which upon action of NPK enriching
microorganisms, can be efficiently used for soil enrichment and
fertility (Lehto et al., 2005). Though the enrichment studies were
carried out for 13 weeks, the data has been presented only upto
6 weeks (Table 3) as there was no remarkable enrichment
observed beyond that.
The pattern of N, P and K enrichment varied depending on the
organism used. Among the various organisms N. muscorum, F. muscicola, A. variabilis, A. fertilissima, C. muscicola, A. lipoferum, A.
chroococcum and mixture considered in the present work, maximum nitrogen (1.3%, w/w) and significant P (0.68%, w/w) and K
(0.8%, w/w) enrichment were found in residue inoculated with A.
variabilis as compared to other organisms. A. variabilis was found
to have high potential in enriching the potato waste which
increased N, P and K by 7.66, 21.66 and 15 fold respectively
(Fig. 3). It was reported that in Anabaena PCC7120, the vegetative
cells supply glutamate to heterocysts, which convert it to glutamine and other amino acids and in return vegetative cells obtain
fixed nitrogen in the form of amino acids from heterocysts (Kumar
et al., 2010) which might be the plausible reason for nitrogen
enrichment. The phosphorous enrichment in the substrate inoculated with the cyanobacteria is due to the conversion of inorganic
phosphorous present in the substrate to soluble form of phosphorous by the action of organic acids (Hariprasad and Niranjana,
2009) released from the phosphorous solubilizing cyanobacteria.
Similar results were reported in other gram negative phosphorous
solubilizing bacteria by Ranjan et al. (2013). Conversion of immobilised potassium into soluble form by the action of organic acid
formed by microflora may contribute for the potassium enrichment of the potato waste. This is in accordance with that reported
by Anthoni (2000) and Rani et al. (2013) in which Bacillus sp. was
employed for solubilization of soil potassium.
A. fertilissima, F. muscicola and Azospirillum lipoferum inoculated
residue showed maximum enrichment in the 5th week and mixture inoculated residue in the 4th week. Thus, maximum NPK
enrichment was observed in the potato waste within 6 weeks of
incubation period. The less encouraging enrichment of residue
treated with mixed culture after 4 weeks is due to the negative
allelopathic interaction among various cyanobacteria in the mixture. For instance, nostocyclamide produced by Nostoc was identified as an efficient anticyanobacterial metabolite (Riley and
Chavan, 2007) which had a pronounced effect on the morphology
of Anabaena. Nostoc spongiaeforme produces a violet pigment nostocine A, which inhibits the growth of many cyanobacteria
(Hirata et al., 1996; Maheep, 2014). The secondary metabolite fischerellin (FsA), isolated from F. muscicola, strongly inhibited the
electron flow in photosystem II in other cyanobacteria (Gantar
et al., 2008).
The N:P:K in A. variabilis enriched residue was found to be
approximately 2:1:1. It was reported that cyanobacterial nitrogen
fixation is essential in the cultivation of rice and for rice-field fertility (Anand and Pereira, 2011). According to NAAS Report

Table 3
Nitrogen, phosphorus and potassium content of potato waste during enrichment
process.
Organism

Week

N% (w/w)

Anabaena variabilis

1
2
3
4
5
6

0.17 0.03
0.23 0.07
0.29 0.05
0.38 0.04
0.89 0.3
1.30 0.3

P% (w/w)
0.07 0.01
0.07 0.02
0.18 0.03
0.27 0.03
0.49 0.04
0.68 0.05

K% (w/w)
0.07 0.016
0.07 0.02
0.10 0.03
0.29 0.02
0.53 0.04
0.8 0.02

Aulosira fertilissima

1
2
3
4
5
6

0.16 0.02
0.18 0.06
0.23 0.08
0.26 0.01
0.84 0.3
0.72 0.03

0.05 0.01
0.05 0.01
0.06 0.02
0.13 0.02
0.32 0.03
0.26 0.02

0.05 0.01
0.07 0.01
0.09 0.01
0.15 0.02
0.36 0.03
0.26 0.02

Nostoc muscorum

1
2
3
4
5
6

0.20 0.01
0.49 0.01
0.78 0.3
0.79 0.2
0.93 0.4
1.20 0.3

0.06 0.01
0.06 0.08
0.15 0.07
0.33 0.05
0.58 0.2
0.74 0.1

0.06 0.01
0.08 0.03
0.21 0.02
0.40 0.06
0.68 0.3
0.82 0.5

Cylindrospermum
muscicola

1
2
3
4
5
6

0.18 0.06
0.26 0.01
0.34 0.01
0.34 0.08
0.46 0.09
0.58 0.03

0.06 0.01
0.06 0.01
0.08 0.01
0.12 0.02
0.18 0.03
0.27 0.07

0.08 0.01
0.10 0.01
0.10 0.012
0.21 0.04
0.29 0.02
0.29 0.05

Fischerella muscicola

1
2
3
4
5
6

0.16 0.08
0.28 0.05
0.33 0.07
0.58 0.02
0.60 0.06
0.61 0.07

0.05 0.03
0.05 0.04
0.05 0.01
0.13 0.05
0.24 0.04
0.14 0.02

0.08 0.02
0.08 0.01
0.09 0.02
0.22 0.01
0.28 0.06
0.21 0.06

Azospirillum lipoferum

1
2
3
4
5
6

0.15 0.08
0.2 0.03
0.23 0.1
0.50 0.03
0.56 0.04
0.50 0.03

0.09 0.01
0.07 0.01
0.06 0.01
0.14 0.03
0.16 0.02
0.16 0.03

0.07 0.02
0.06 0.01
0.07 0.013
0.11 0.015
0.28 0.04
0.17 0.02

Azotobacter chroococcum

1
2
3
4
5
6

0.15 0.08
0.17 0.04
0.39 0.02
0.46 0.01
0.67 0.02
0.70 0.03

0.06 0.01
0.05 0.03
0.15 0.07
0.19 0.01
0.23 0.02
0.22 0.06

0.07 0.01
0.09 0.01
0.11 0.02
0.15 0.01
0.23 0.05
0.29 0.03

Mixture

1
2
3
4
5
6

0.15 0.04
0.17 0.03
0.18 0.08
0.19 0.03
0.17 0.04
0.14 0.03

0.05 0.02
0.065 0.03
0.072 0.05
0.08 0.07
0.08 0.08
0.074 0.03

0.09 0.02
0.105 0.04
0.11 0.05
0.12 0.02
0.11 0.05
0.10 0.06

Fig. 3. Increase in nutrients before and after enrichment (mediated by Anabaena


variabilis).
Table 2
Biochemical characteristics of potato waste before enrichment process.
Parameters

Content in potato waste

Nitrogen (% TS)
Potassium (% TS)
Phosphorous (% TS)

0.15 0.05
0.05 0.01
0.03 0.001

(2009), the N:P:K recommendation for rice, potato, sorghum, cotton and wheat field is about 4:2:1 to 4:2:2. Biomanure from potato
waste can be specifically used for enrichment of alluvial and lateritic soils which needs an adequate supplementation of nitrogen.

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A.D. Chintagunta et al. / Waste Management 49 (2016) 320325

Fig. 4. Rough estimate of bioethanol and biomanure production from potato waste (based on the data obtained from a: Agricultural statistics at a glance, 2013; b: Pandey
et al., 2009; c: Guttormsen and Carlson, 1969; d: Data from present study; e: Average value of solid utilized for ethanol production from peel and mash).

3.4. An estimate of bioethanol and biomanure production from potato


waste in India
Based on the outcome of the present study, it is important to
estimate the bioethanol and biomanure production from the generated quantum of wastes from the potato processing industries
in India throughout the year. Fig. 4 represents an estimated potential of ethanol and manure production from the available potato
waste resources.
4. Conclusion
Significant amount of ethanol was produced in short incubation
time by employing co-culture of S. cerevisiae and A. niger to potato
waste through SSF. From the results, it could be concluded that
potato wastes from potato processing plant is an attractive raw
material for ethanol production with simultaneous reduction of
waste thereby circumventing environmental pollution. In addition,
the study also explored the possibilities of utilizing the residue
obtained after ethanol fermentation for further enrichment to be
applied as biomanure. The obtained enriched residue offers an economically viable alternative to chemical fertilizers for accomplishing the ultimate target of increased productivity.
Acknowledgment
Authors acknowledge DST-TIFAC for financial assistance to
carry out this work.
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