Bioresource Technology Reports 18 (2022) 101092

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Bioresource Technology Reports

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Bioethanol from various types of banana waste: A review

Ashish N. Sawarkar a, *, Nikhil Kirti a, Ankita Tagade a, Shyam P. Tekade b
a
Department of Chemical Engineering, Motilal Nehru National Institute of Technology Allahabad, Prayagraj 211004, Uttar Pradesh, India
b
Department of Chemical Engineering, Gharda Institute of Technology, Lavel 415708, Maharashtra, India

A R T I C L E I N F O A B S T R A C T

Keywords: Various types of banana waste (BWs) generated after banana crop cultivation have tremendous potential for
Banana wastes bioethanol production. The present review paper aims to bring together information on various facets such as i)
Pretreatment variation in physico-chemical and biochemical composition of various types of banana waste ii) various pre­
Biochemical conversion
treatment methods, viz. mechanical, chemical, and biological iii) acid hydrolysis and enzymatic hydrolysis iv)
Fermentation
Bioethanol
fermentation mechanism and role of microbes in banana wastes conversion v) various fermentation schemes for
bioethanol production from banana wastes, coherently on a single platform based on reported literature. Analysis
revealed that novel pretreatment techniques such as organic acid pretreatment and electrical pretreatment
techniques such as microwave and ultrasonic are making substantial progress. Robust statistical approaches are
proven to be effective as regards to process parameters optimization. Finally, current challenges along with
research needs for making further inroads in the production of bioethanol from various BWs have also been
deliberated.

1. Introduction Bioethanol has piqued widespread interest on a global, national, and

regional scale. Recently, India has set a bold target of 20% ethanol-
Phenomenal growth in industrialization, lifestyle modernization, blending with gasoline by 2023–24 (Business Standard Report, 2021),
and vehicular population, all over the world, has resulted in a sub­ which is currently at about 9%. Bioethanol has high octane number
stantial rise in the energy demand. Currently, fossil-based fuels satisfy (106–114) (Aditiya et al., 2016), higher oxygen content (34.7%)
approximately 80% of the world's energy demand, with petroleum ac­ (Masum et al., 2014) and low boiling point in comparison with gasoline.
counting for 33% of the total energy supply. At the same time, in view of Further, it has high heat of vaporization (840 kJ/kg) than gasoline (305
the fluctuating crude oil prices accompanied by increasing environ­ kJ/kg), ensuring better volumetric efficiency of ethanol blend than pure
mental concerns pertaining to the use of fossil fuels, the demand for gasoline and therefore relatively high power output (Lynd et al., 1991).
biofuels is progressively increasing all over the world (Gokul et al., Bioethanol is considered eco-friendly as its high octane number prevents
2019; Gajera et al., 2020; Singh et al., 2021a; Singh et al., 2021b). engine knocking and facilitates early ignition (Nwufo et al., 2013).
Among various renewable energy sources, biomass is gaining in Because of its higher oxygen content, it endures for cleaner combustion
increasing attention because it is considered the most reliable and and lowers soot and hydrocarbons (HC) emissions (Krishnan et al.,
environment-friendly energy source with a host of product slates 2020). With lower energy content than petrol, it helps in the reduction of
(Tagade et al., 2021). Biofuels offer appealing solutions for lowering emissions of CO2 by 90% and SO2 by 60%–80% (Halder et al., 2019).
greenhouse gas emissions, reducing dependence on fossil oil, confront­ The various raw materials utilized for the production of bioethanol
ing energy security concerns, enhancing agricultural economies, and are categorized into four types, based on the source (feed) for its pro­
improving the sustainability of the global transportation system (Singh duction (Azad et al., 2015). First generation bioethanol is derived from
et al., 2021c). edible crops such as rapeseed, sunflower, oilseeds, potato, etc. whereas
Among the several biofuel substitutes, bioethanol is the most sought- second generation emanated from lignocellulosic biomass (Binod et al.,
after and widely produced biofuel (Passoth and Sandgren, 2019) that is 2019). Third generation bioethanol is produced from high carbon
foreseen to have a potential future in addressing today's global energy embedded algal biomasses such as microalgae (Azmah et al., 2016;
dilemma and deteriorating environmental quality (Kundu et al., 2021). Mathimani and Pugazhendhi, 2019; Patle et al., 2021). Although the

* Corresponding author.
E-mail address: [email protected] (A.N. Sawarkar).

https://doi.org/10.1016/j.biteb.2022.101092
Received 2 March 2022; Received in revised form 14 May 2022; Accepted 16 May 2022
Available online 20 May 2022
2589-014X/© 2022 Elsevier Ltd. All rights reserved.
A.N. Sawarkar et al. Bioresource Technology Reports 18 (2022) 101092

production of bioethanol from algae is a promising technology, there are 2. Banana wastes
significant limitations that need to be addressed before biofuel can be
commercialized and produced on a wide scale. Some of the most critical 2.1. Types of banana wastes and their typical applications
issues include profound knowledge regarding control of carbohydrate
mechanism of microalgae, biomass harvesting, low yield, production of Banana is one of the most important monocotyledonous annual fruits
algal biomass in sufficient quantities and high capital and operational grown in tropical and subtropical parts. It comes from the family of
costs of microalgal biomass bioreactors. Genetically altered microor­ Musaceae. Bananas are produced in around 130 countries. It is the
ganisms such as cyanobacteria, yeast, microalgae, and fungus are world's oldest farmed crop and one of the tallest herbaceous plants with
employed as raw materials to manufacture sustainable bioethanol as the a pseudostem (Gupta et al., 2022). The socio-economic significance in
fourth generation biofuel (Saha et al., 2019). The fourth-generation banana cultivation in terms of food security and as a source of export
biofuels are considered to be in their embryonic stage since the large revenue is quite palpable in some countries, which underlines the
scale production of such feed materials is currently not feasible due to importance of this fruit. Globally, approximately 115 million tons of
health and ecological risks of cultivating genetically modified microbes, bananas are produced per year. India is the largest producer of banana in
issues regarding legality, limited biomass output, and high the world (Kirti et al., 2020; Singh et al., 2020c) with a production of
manufacturing costs. around 32 million tons per annum, which makes about 28% of overall
From among various generations, lignocellulosic biomass which banana production in the world. Other prominent banana producing
typically involves agricultural and forest residues as feedstocks have countries include China, Indonesia, Brazil, Ecuador and the Philippines.
received tremendous attention for the production of bioethanol, mostly Fig. 1 depicts the percentage share of top banana producing countries.
because of the fact that it is not entangled in food vs. fuel debate, Banana plant bears a bunch of fruit once in a lifespan (Balogun et al.,
associated with first-generation biofuel feedstocks (Morales et al., 2018; Singh et al., 2022), thereby generating a considerable quantity of
2021). In particular, biofuels from agricultural residues has been a agro residues both at the time of harvesting as well as after processing of
sustained area of research. Globally, India is one of the foremost fruits (Tan et al., 2019). For one ton of bananas obtained, approximately
agricultural-based economies and livelihood of ~75% of its populace 100 kg of fruit is rejected and about 4 tons of wastes including 160 kg
banks on agriculture (Singh and Sawarkar, 2020) and consequently stalks, 440 kg skins, 480 kg leaves, and 3 tons pseudostem are produced
generates about 350 million tons of agricultural waste biomass annually (Fernandes et al., 2013). Banana pseudostem has trunk like structure
(Singh et al., 2020a). Moreover, The National Policy on Biofuels–2018, formed by tightly packed overlapping leaf sheaths. It provides support to
Government of India has laid significant emphasis on the production of the plant and mostly consists of water. Every pseudostem is replaced by
biofuels from agricultural waste biomass (Singh et al., 2020b; Kirti et al., another pseudostem after bearing fruit once. Banana leaves are gener­
2022). As a result, bioethanol production has got further impetus for ally 30–60 cm wide and up to 2 m long. Banana peels are outer covering
efficient agricultural residue utilization. for the banana fruit whereas rachis is the flower stalk.
After a close scrutiny of the reported literature, it was revealed that Banana and its agricultural waste have wide range of applications.
investigations pertaining to bioethanol production from rice straw They are used in food sector, paper and pulp sector, energy sector, fibre
(Binod et al., 2010), wheat straw (Talebnia et al., 2010; Ingrao et al., and film, and bioplastic (Tock et al., 2010; Ulloa et al., 2004; Padam
2021), corn stover (Zhao et al., 2018), sugarcane bagasse (Huang et al., et al., 2014; Gumisiriza et al., 2017; Rana et al., 2018; Sogi, 2020).
2020) have been presented in the form of a review paper, however, the Banana produced fibres are comparable to fibres obtained from fibrous
studies associated with bioethanol production from various banana commodities in terms of physical strength and cellulose content and
associated wastes have not been put together coherently on a single have been characterized from their fruit stalk, pseudostem and leaves
platform. It may be pointed out that banana is an extensively cultivated (Bello et al., 2014). Banana fibres and banana microfibrils are produced
fruit crop in the world and India is the largest producer of banana in the by treatment of banana waste with alkaline pulping and steam explosion
world and in turn banana residues which majorly comprises pseudos­ (Ibrahim et al., 2010) and these are used to process composite material
tem, leaves, rachis, and peel. The production of bioethanol from various by using polyethylene and polypropylene as the polymeric matrix
banana wastes has recently received much attention. As per the Scopus (Chattopadhyay et al., 2010; Ibrahim et al., 2010). The fibres obtained
database, in the last decade (2010− 2020), the number of publications from banana pseudostem have been used to reinforce epoxy composites
concerning the topic observed an ever-increasing trend. The total and raw materials for textile industry for production of traditional
number of publications reporting the subject matter was 7 till 2010 handicrafts and clothes (Padam et al., 2014). The pith from banana
which increased to 143 in 2020 and reached to 177 in 2021. According
to the Scopus database, the top academic journals publishing the work
on bioethanol production from banana wastes are Renewable and Sus­
tainable Energy Reviews, Bioresource Technology, Journal of Cleaner
other
Production, and Industrial Crops and Products. Studies reported during 8%
India
2008 to 2021 pertinent to the chosen topic are considered in this review Ecuador
paper. The aim of this review paper is to present a detailed overview of 10% 28%
various facets of production of bioethanol from a variety of banana
wastes, bottlenecks involved, and recommendations for future work.
Section 2 deliberates on worldwide production of banana including China
various types of banana waste and their conventional applications. It
18%
also throws light on the physico-chemical properties and biochemical
composition of various types of banana waste. Section 3 deals with Indonesia
various pretreatment methods which have been employed for different 15%
banana wastes. Section 4 deals with actual steps involved in the bio­ Brazil
ethanol production from a variety of banana wastes comprising, acid 12%
hydrolysis and enzymatic hydrolysis, role of microorganisms in the
conversion, and various fermentation schemes for bioethanol produc­ Philippines
tion from banana wastes. Challenges and possible solutions are dis­ 9%
cussed in Section 5 and research needs and future directions are
discussed in Section 6. Fig. 1. Percentage share of top banana producing countries.

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A.N. Sawarkar et al. Bioresource Technology Reports 18 (2022) 101092

stems is used in spices. Furthermore, subsequent to decomposition, ba­ Table 2

nana pseudostem is used for mushroom cultivation together with other Biochemical analysis of various reported banana biomass wastes.
agricultural wastes such as wheat straw. Banana pseudostem has also Banana Biochemical analysis (wt%) Reference
been used for paper production. Banana waste pulp papers are more waste
Cellulose Hemicellulose Lignin Extractives
water resistant and stronger than wood-pulp and can be combined with
bamboo pulp in making greaseproof paper (Jacob and Prema, 2008; Banana 9.9 41.38 8.9 – Kabenge et al.
peel (2018)
Goswami et al., 2008). Temperature, pH, and pulping time are the 12.17 10.19 2.88 – Pathak et al.
important factors deciding the yield of banana waste trunk pulp (2017)
(Ogunsile et al., 2006). Banana 35.58 23.46 10.58 – Kabenge et al.
Banana peel can act as binder and base material, which can be used leaves (2018)
Semi 26.7 25.8 17 Fernandes
in fabrication work. Banana peels are used to make briquettes with –
dried et al. (2013)
lower burning rate than other agro-residue briquettes such as rice husk, Banana
sawdust, coconut fibre, peanut shell, and palm fibre with equivalent leaves
strength (Chin and Siddiqui, 2000). Apart from briquetting, banana Wet dried 53.5 28.5 15.4 – Fernandes
wastes are utilized as an efficient biofertilizer for enhancing agricultural banana et al. (2013)
leaves
productivity and soil fertility. Lim and Matu (2015) prepared bio- Banana 38.48 25.36 5.77 – Kabenge et al.
fertilizer from five types of agro waste; watermelon, papaya, pine­ pseudo (2018)
apple, citrus orange, and banana and it was found that biofertilizer from stem 38.36 29.05 17.81 14.78 Kumar et al.
banana contains highest amount of potassium (3.932 g K/L) and has pH (2019)
25 25 15 Kumar et al.
value of 4.52 which makes banana agro-waste suitable to produce bio­ –
(2020)
fertilizer for plants. Further, banana peels can also be converted into Rachis 32.1 8.8 19.0 Velásquez-
methane by rapid fermentation of peels in a plug-flow biogas reactor arredondo
(PFBR) (Chanakya et al., 2009). In addition, banana peels can be con­ et al. (2010)
verted to renewable fuels by enzymatic hydrolysis followed by 23 11.2 10.8 18.2 Guerrero and
Muñoz
fermentation assisted with microbes (Manikandan et al., 2008; Saxena (2018)
et al., 2009). Traditionally, banana leaves are used to serve meals and as 33.2 8.4 18.2 Villa-vélez
a covering for steamed food preparation. Banana leaves are also et al. (2015)
employed in a variety of ceremonial occasions, and their use is
expanding due to cultural diversity. However, it inevitably turns into
source for fermentable sugar as cellulose has higher degree of biodeg­
waste after it has fulfilled its purpose.
radation than hemicellulose and lignin (Pérez et al., 2002). Further, low
lignin concentration of banana wastes (<20%) significantly reduces
2.2. Physico-chemical and biochemical composition of various banana lignocellulose hydrolysis time, as well as energy consumption and pro­
wastes duction costs (Bhushan et al., 2019). Lignin and moisture content play
important role in selection of biomass for energy production. Low lignin
Table 1 shows proximate and ultimate analysis of various banana and low moisture are preferable for better energy production (Bhatia
wastes. It can be envisaged that the various banana wastes contain et al., 2017). Meng et al. (2019) studied the chemical composition
moisture, volatile matter, ash and fixed carbon in different proportions. changes during the liquefaction process of banana pseudostem and
Moisture content is an essential element that impacts the performance of estimated kinetic parameters of raw banana pseudostem and its three
enzymatic hydrolysis and fermentation. To improve conversion effi­ main components. Cellulose was found to have lowest reaction rate and
ciency, the water content of the material should be in the desirable required maximum activation energy of 41.32 kJ mol− 1 compared to
range. As can be seen from the ultimate analysis, the carbon and oxygen hemicellulose (18.74 kJ mol− 1) and lignin (26.14 kJ mol− 1).
content put together in the banana agro-waste is about 90%, with
pseudostem having the maximum percentage of these two. The 3. Pretreatment of banana wastes for conversion to bioalcohol
biochemical composition of various banana wastes has been shown in
Table 2. A comprehensive biochemical analysis is imperative to estab­ Pretreatment of banana wastes is of critical importance in the overall
lish actual fractions of different polymers and to build an enzyme conversion of lignocellulosic-derived sugars to biofuels (Rezania et al.,
cocktail for effective hydrolysis (Liu et al., 2021). It was revealed from 2020), especially for the production of bioethanol. The primary con­
Table 2 that the banana wastes contain significant proportions of cel­ stituents of banana lignocellulosic wastes are cellulose, hemicellulose
lulose and hemicellulose. Cellulose rich banana wastes are a potential and lignin. The linkage between the individual (intra-polymer linkages)

Table 1
Proximate and ultimate analysis of various reported banana biomass wastes.
Banana waste Proximate analysis (wt%) Ultimate analysis (wt%) Reference

Moisture Volatile matter Ash Fixed carbon C H N S O

9.80 85.26 5.01 0.07 40.24 6.14 1.3 0.098 52.22 Pathak et al. (2017)
Banana peel 11.56 88.02 9.28 2.7 35.65 6.19 1.94 – 45.94 Kabenge et al. (2018)
11.56 77.84 8.21 2.39 35.65 6.19 1.94 – 45.94 Ighalo and Adeniyi (2019)
6.67 83.35 9.05 7.60 38.57 6.44 2.45 – 43.49 Kabenge et al. (2018)
Banana leaves
6.67 77.79 8.45 7.09 38.57 6.44 0.8 – 43.49 Ighalo and Adeniyi (2019)
Wet dried banana leaves 74.7 41.3 12.2 46.5 15.90 9.20 1.4 0.05 73.40 Fernandes et al. (2013)
Semi dried banana leaves 8.3 78.8 8.7 12.5 43.5 6.3 1.3 0.2 48.7 Fernandes et al. (2013)
6.67 74.33 11.67 7.33 33.09 2.94 0.94 – 63.03 Kumar et al. (2020)
7.98 82.29 8.61 1.12 33.46 6.44 0.8 0.04 49.94 Ighalo and Adeniyi (2019)
Pseudo stem
7.98 89.43 9.36 1.21 33.46 6.44 0.8 – 49.94 Kabenge et al. (2018)
– 76.5 8.64 14.86 42.00 5.62 1.08 – 51.30 Kumar et al. (2019)
Rachis – 76.0 19.0 5.0 33 4.3 1.9 0.3 60.5 Islam et al. (2019)

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A.N. Sawarkar et al. Bioresource Technology Reports 18 (2022) 101092

and different components (inter-polymer linkages) of lignocellulose Table 3

complex is provided by four distinct bonds, viz. ether bonds, carbon to Summary of advantages and disadvantages of various pretreatment methods.
carbon, hydrogen bond, and ester bond. The pretreatment of banana Pretreatment Advantages Disadvantages
wastes involves breaking of these bonds, which alters the structure of
Physical Grinding Reduces cellulose High power and
lignocellulosic biomass, making cellulose more accessible to enzymes crystallinity and energy requirement
(Krylova et al., 2008). Pretreatment reduces the degree of crystallinity polymerization
for subsequent hydrolysis and fermentation process (Aditiya et al., degree
2016). It also increases the porosity and reactivity of banana wastes and Milling Decreases degree of Equipment costly
polymerization with high
enhances the overall yield of biofuel (Kumar et al., 2009; Saritha et al., Increases specific maintenance
2012). Further, the pretreatment may also result in the release of some surface area along High power and
fermentable sugars by depolymerizing the hemicellulose component of with pore size energy prerequisite
the lignocellulose. Researchers have also employed the fundamentals of Irradiation Easy operation Expensive
Reduction or High energy
response surface methodology to optimize the pretreatment models for
avoidance of consumption
converting lignocellulosic biomass into fermentable sugars and then to inhibitory substances
bioethanol (Manmai et al., 2021). The selection of pretreatment method Microwave Highly capable to Lack of large scale
for the biomass depends on various factors such as composition and assisted break recalcitrant equipment
characteristics of residue, energy and power consumption (Putro et al., structure of High consumption
lignocellulosic of electricity for
2016; Bhatia et al., 2018; Lemões et al., 2018). Various pretreatment biomass microwave
methods for banana wastes include mechanical processing, chemical Increases enzymatic generation.
exposure, physicochemical and physical pretreatment, biological pre­ susceptibility
treatment or a combination of these pretreatment methods. Table 3 High heating
efficiency
summarizes advantages and disadvantages of various pretreatment
Easy operation
methods for banana wastes for the production of bioethanol. Requires short time
Physicochemical Steam Efficient in breaking Generation of
3.1. Mechanical pretreatment explosion recalcitrant nature of cellulose inhibitory
lignin by increasing compounds that
surface area and hampers enzymatic
Mechanical pretreatment process includes physical forces such as removing pentose hydrolysis
drying, chopping, milling and grinding to the biomass. These physical sugar Partial
forces lead to the increase of surface area and reduction in particle size, Hemicellulose hemicellulose
degree of crystallinity and degree of polymerization (Tassinari et al., solubilization degradation
High recovery of
1980). Primary mechanical pretreatment employs milling and grinding
xylose
to reduce particle size. Palacios et al. (2017) compared pulverized ba­ High yield of glucose
nana peels with unmilled peels for cellulose yield and it has been re­ and hemicellulose
ported that pulverized banana peels result in higher cellulose yields. occurring in two step
Mechanical pretreatment can be combined with other pretreatment method
Cost effective
modules for making process more efficient and economical. Thermo- CO2 explosion Not toxic, not Requires high
mechanical pretreatment is one of the common pretreatment module flammable pressure
practices commonly with steam. Banana trunks are used to produce Increase cellulose
binderless particleboard and steam pretreatment improves the me­ digestibility
Absence of the
chanical and physical strength of particleboard and provides dimen­
inhibitory
sional stability (Nadharia et al., 2019). compounds
Easy recoverable
3.2. Chemical pretreatment after extraction
Cost effective
Liquid hot Green technology as High energy and
Chemical pretreatment is one of the most important pretreatment water it eliminates the water requirement
methods. It facilitates the breaking of a rigid lignocellulosic matrix with/ usage of chemicals Low sugar
without hemicellulose and improves biodegradability. It renders ligno­ Relatively high concentrations
cellulosic biomass more amenability to rapid hydrolysis, resulting in xylose recovery Generation of
Economical degrading
higher monomeric sugar yields. It also decreases degree of polymeri­
pretreatment compounds which
zation and crystallinity of the cellulose components. Chemical methods inhibits microbial
such as lime, acid, alkali, ammonia fibre explosion, sulfur dioxide ex­ growth
plosion, and ionic liquids have been used for pretreatment of biomass Ammonia fibre Simple, short High cost for large
with organic acids (acetylsalicylic acid, oxalic acid, salicylic acid) as expansion processing time amount of
Improves proximity ammonia
catalyst (Behera et al., 2014). Chemicals with acid or alkali are used to of cellulose and High temperature
fractionate complex cell wall matrix and depolymerize cellulose content cellulose enzyme and pressure
(Putro et al., 2016). Guerrero et al. (2018) followed acid catalyzed steam High digestibility of requirement
explosion pretreatment of banana pseudostem and rachis with different lignocellulosic Less efficient
feedstock with high towards high lignin
conditions. Pseudostem was treated at 177 ◦ C for 5 min of residence time
enzymatic hydrolysis content biomass
and 2.2% H2SO4 (v/v), presenting a high overall glucose yield of 91.0% yield
and rachis at 198 ◦ C for residence time of 5 min and 1.5% H2SO4 (v/v) Ammonium salt
with an overall glucose yield of 87.1%. In another study, Gabhane et al. residue is easy to
(2014) studied the compositional analysis of banana waste using mi­ recover and can be
utilized as microbial
crowave, autoclave and ultrasonic pretreatment. Powdered banana nutrient
parts were treated in 100 mL of 5% (v/v) H2SO4 or 1% (w/v) NaOH The hydrolysate can
solutions. Temperature for microwave and autoclave assisted pretreat­ (continued on next page)
ment were 180 ◦ C and 121 ◦ C respectively. Ultrasonic pretreatment was

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A.N. Sawarkar et al. Bioresource Technology Reports 18 (2022) 101092

Table 3 (continued ) imparted at 25 kHz frequency and power of 150 W. Thakur et al. (2012)
Pretreatment Advantages Disadvantages pretreated the banana and wheat stem for enzymatic hydrolysis and
fermentation. 1 N NaOH or 1 N H2SO4 was incubated with 10% slurry of
be utilized without
additional disposal.
banana or wheat stem at room temperature. Guo et al. (2018) employed
Chemical Concentrated High conversion rate Toxic, corrosive acid pretreatment for banana peels using different organic acids such as
acidic of sugar High cost oxalic acid and tartaric acid at different temperatures and reaction
Hydrolysis carried times. According to their findings, pretreatment with 0.2% tartaric acid
is relatively slow
resulted in the maximum conversion (48.41%) of reducing sugar and
High operational
and maintenance 0.2% oxalic acid resulted in maximum conversion (46.39%) of reducing
cost sugar, at 100 ◦ C and for a reaction time of 65 min. Costa et al. (2018)
Necessitates reported a number of chemical pretreatments for banana rachis for
equipment with lignin degradation. Organic solvents (organosolv) have been chosen for
acid corrosive
resistance
pretreatment as it removes lignin and hemicellulose by solubilization
Need of acid and extraction. Cellulose is more susceptible to enzymatic degradation
recovery as compared to hemicellulose and lignin (Pérez et al., 2002). Acid pre­
Dilute acidic Fast reaction process Generation of treatment provides high yield of sugar as it solubilizes the hemi­
Less corrosion as degradation
celluloses fraction from biomass but may cause corrosion to the waste
compared to products
concentrated acid Low concentration carrying pipeline. Attempts have also been made to investigate the
of reducing sugar utilization of different organic acids in the acid pretreatment. In this
High temperature line, Guo et al. (2018) explored retreatment of banana peels by
and pressure employing tartaric, oxalic, and citric acid and compared it to the sulfuric
Alkaline Decreases degree of High cost of alkalis
polymerization Salt absorption into
acid pretreatment. The pretreatment experiment was conducted sepa­
Decreases biomass while rately with 2 g of peel powder immersed in concentration of 0.2% of
crystallinity of pretreatment acid, maintained at 80 ◦ C for 60 min on an orbital shaker at 50 rpm.
cellulose reactions Tartaric acid was found to achieve the highest conversion rate of
Effectively breaks
reducing sugar with 48.41%, while oxalic acid achieved second
recalcitrant lignin
structure via swelling maximum conversion with 46.39%, followed by sulfuric acid and citric
of biomass acid. Furthermore, using life cycle assessment (LCA) technique, Santiago
Enhances residual et al. (2022) conducted a comparative environmental analysis of several
polysaccharide valorization scenarios of banana peel waste to bioethanol. According to
reaction
their findings, valorisation of banana peel for bioethanol with oxalic
Favourable for
enzyme hydrolysis acid pretreatment is the most environmentally benign method. Reddy
End residue et al. (2019) investigated sequential alkali and acid pretreatment for
(generally cellulose) banana crop residue. According to their investigation, the sequential
can be utilized in
pretreatment with 1% alkali and 1% acid facilitated lignin solubilization
various application
Requires low of 56% and 61%, respectively. Hossain et al. (2019) reported glucose
temperature and production from the banana stem with two pretreatment methods for
pressure bioethanol production. They employed acid hydrolysis by diluting sul­
Wet oxidation Enhanced abstraction Stringent reaction furic acid (5.614 g/L, 90 min, and 0.5 M H2SO4) and enzymatic hy­
of lignin conditions
drolysis (40.61 g/L,72 h) by cellulase enzymes for glucose production
Low formation of High pressure and
inhibitor temperature which later on was fermented in the presence of Saccharomyces cerevisiae
Organosolv Extraction of high Expensive solvents yeast for bioethanol production. They also studied the energy generation
quality lignin which Efficient control of from bioethanol and reported lower CO2 (97,161 kg/y) and SO2 (513
can be utilized for the pretreatment
kg/y) emissions from diesel. Alkali pretreatment solubilizes more lignin
range of applications because of volatility
High purity cellulose of solvents
fraction, does not require high temperature and pressure but requires
separation with Solvents from the more time and high dosage of alkali such as calcium, potassium, sodium
minor degradation system causes and ammonium hydroxide (Shimizu et al., 2018; Bhushan et al., 2019).
Readily solvent certain effects on Tiwari et al. (2019) achieved recovery of sugar by microwave assisted
recovery fermentation
alkali (NaOH) pretreatment on banana peels and showed effect of mi­
Low input Solvent drainage
has an influence on crowave power and irradiating time on composition of sugar. Maximum
the environment concentration of reduced sugar 0.561 g/g of dry banana peels is reported
Biological Environment friendly Low hydrolysis rate at 600 W microwave power for 2 min. Similarly, a study was reported on
Effective in Time consuming
microwave-assisted alkali pretreatment of banana pseudostem. 8%
degradation of lignin
Mild condition
NaOH ratio was used for pseudostem pretreatment at 90 ◦ C for 8 min
Low consumption of (Chittibabu et al., 2014). Reddy et al. (2010) employed 1% NaOH and
energy 1% H2SO4 for the pretreatment of banana leaves after steam explosion
No usage of via autoclave at 121 ◦ C for 15 min. According to their investigation, with
chemicals
alkali pretreated banana leaves, a maximum ethanol production of 3.82
Combined Effective lignin and High energy
hemicellulose demand g/l was achieved, with an ethanol yield of 0.41 g/g of cellulose degra­
degradation Special equipment dation. Shimizu et al. (2018) studied the effect of acid (H2SO4), alkali
Better enzymatic design (NaOH) and peroxide (H2O2) pretreatment on banana pseudostem for
hydrolysis
bioethanol production. It has been reported that alkaline and peroxide
pretreatment strongly reduced hemicellulose (19.62%) to 4.38 and
8.68%, respectively and lignin content (17.26%) to 7.65% and 7.17%,
respectively. Cellulose content (60.84%) was found to increase to
75.48% and 74.37% with the aid of alkaline and peroxide pretreatment,

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respectively. Acid pretreatment module, on the other hand, completely paths. The conversion route is very crucial for the bioalcohol production
removed hemicellulose but increased lignin content from 17.26% to from biomass. The essence of all the conversion routes for bioalcohol
39.99%. Legodi et al. (2021) compared the cellulose production from production from banana wastes lies in destroying the crystalline struc­
three different pretreatment methods NaOH, H2SO4, and hot water and ture of lignocelluloses by removing as much lignin as possible and
reported 52%, 48%, and 25% cellulose, respectively after pretreatment. separating hemicellulosic and cellulosic components. Fig. 2 shows
The highest bioethanol (17.6 g/L) was produced via alkaline pretreated schematic representation of various steps involved in biochemical con­
banana stem using Saccharomyces cerevisiae UL01 at 30 ◦ C and 100 rpm version of banana wastes to bioethanol.
converting 51% of glucose. They also reported a faster hydrolysis rate in
hot water than acid and alkaline pretreatment. Microwave assisted
pretreatment, ultrasonic assisted and electrical pretreatment are basi­ 4.1. Biochemical conversion of banana wastes to bioethanol
cally considered under physical pretreatment module and physico­
chemical pretreatment includes CO2 explosion, steam explosion, Biochemical conversion is touted to be an eco-friendly and sustain­
ammonia fibre expansion, and oxidative pretreatment (Kumar and able approach that employs biological agents, viz. bacteria, fungus or
Sharma, 2017). Albarelli et al. (2011) studied the effect of supercritical their enzymes for bioethanol production. Biochemical conversion has its
carbon dioxide on banana peels for adsorptions of heavy metals. Su­ own advantages such as less energy input, self-sustained, suitability for
percritical CO2 was used for solvent extraction as well as biomass high moisture content biomass (Gumisiriza et al., 2017). Subsequent to
treatment. Ultrasound pretreatment uses the principle of cavitation. pretreatment, hydrolysis and fermentation are the two significant steps
High temperature and pressure are produced after collapsing of micro­ in the biochemical production of bioethanol from banana wastes (Ghosh
scopic vapor bubble creating hot spot in cold liquid. Villa-vélez et al. et al., 2017). Further, rate of bioconversion depends upon properties of
(2015) studied the effect of concentration, pH, ultrasonic power and the substrate, microbial agents and process conditions.
time for pretreatment of banana flower stalk. They showed the effect on
total sugar and reduced sugar yield and reported 98.1–556.2 mg/g of 4.1.1. Hydrolysis of banana wastes
total sugar yield at 300–1500 W. Alkali pretreatment reported less The process of conversion of biomass biopolymers to fermentable
process time (24 h) than that of acid (84 h) and microwave pretreatment sugars is known as hydrolysis or saccharification. The hydrolysis out­
(84 h). Alkali pretreatment has a higher amount of lignin removal at low comes are typically determined by the type and properties of the
temperature and pressure whereas, in acid pretreatment, the tempera­ biomass such as concentration, crystallinity, pretreatment methods, pH,
ture is inversely proportional to the acid concentration. Further, high temperature, mixing, and hydrolysis conditions (Aditiya et al., 2016).
temperatures of the acidic pretreatment result in a release of some un­ The hydrolysis of banana wastes into fermentable sugars is a critical
desired compounds like furfural, acetic acid, and 5-hydroxymethylfurfu­ stage that greatly influences overall process efficiency. Primarily, hy­
ral (HMF). At modest concentrations (<1 g/L), these chemicals can drolysis of banana wastes is accomplished by two techniques. One
inhibit cellulase enzymes and fermentation organisms. Hence removal method employs the use of acids as catalysts, while the other makes use
of inhibitory compounds remains a challenge for acid pretreatment. of enzymes (Guo et al., 2018).

3.3. Biological pretreatment 4.1.1.1. Acid hydrolysis. Acid hydrolysis is mainly accomplished by
dilute or concentrated solutions of sulfuric or hydrochloric acid. The
Biological pretreatment uses microorganisms and enzymes for the production of chemicals such as furfural, hydroxymethylfurfural (HMF),
degradation of biomass. It is an eco-friendly approach with minimal and acetic acid, which act as inhibitors to the microorganisms that
energy input and mild environmental conditions (Mielenz, 2015). Bio­ generate ethanol from sugars are most commonly encountered bottle­
logical pretreatment depends upon strain selection, incubation condi­ necks (Zabed et al., 2016). Corrosion of the equipment due to high acid
tions such as temperature, pH, substrate load, and agitation. It involves concentrations, as well as the need for acid recovery or neutralization, is
microorganisms such as fungi which include brown rot, white rot, and another key disadvantage of this approach. De Souza et al. (2013)
soft rot for degradation of lignin and hemicellulose. White and soft rot investigated acid hydrolysis of banana wastes, particularly banana
attacks both lignin and hemicellulose whereas, brown rot attacks only pseudostem. According to their findings, dilute acid hydrolysis for ba­
lignin. Class II peroxidases enzymes such as manganese peroxidase, nana pseudostem converted cellulose to 36% of glucose at optimum
lignin peroxidase, and versatile peroxidase; Cellulase enzymes such as conditions of 2% H2SO4 and 15 min residence time at 120 ◦ C.
species of Cellulomonas, Clostridium, and Streptomyces, and fungi like
Aspergillus, Humicola, Phanerochaete and Trichoderma have been used 4.1.1.2. Enzymatic hydrolysis. Enzymatic hydrolysis is a biologically
for degradation of lignin. Aspergillus species has been reported as the derived, environmentally benign, and efficient technique of hydrolyzing
most employed fungi for biological pretreatment of banana residues both cellulose and hemicellulose into fermentable sugars (Keshwani and
(Ghasemzadeh et al., 2017; Bhushan et al., 2019). Ingale et al. (2014) Cheng, 2009). It is performed by employing cellulase which is a group of
employed Aspergillus ellipticus and Aspergillus fumigatus under co-culture various cellulytic enzymes that work together to break down cellulose to
fermentation of banana pseudostem. These microbes were used to produce oligosaccharides, cellobiose, and glucose. Complete hydrolysis
degrade lignin and hemicellulose for producing reduced sugar. Singh of cellulose requires synergistic action of at least three enzymes to break
et al. (2015) produced bioethanol with yield 6.287% (w/w) after pre­ down cellulose and provide a usable energy source in the form of
treatment of banana peels with Aspergillus niger. Ghosh et al. (2019) glucose. An efficient cellulase is supposed to degrade the cellulose in its
studied biological pretreatment of banana pseudostem using semi- crystalline form while also being resistant to acidic pH (4, 5) and stress
defined synergistic microbial consortium consisting of baker's yeast situations (Balat, 2011). The utilization of enzymes in cellulose hydro­
and the native fungal sp. of banana pseudostem for pyro-oil generation. lysis is efficient in terms of high selectivity and catalytic action, low
The pretreatment occurred at various pretreatment times, viz. 24 h, 36 energy requirement, mild process conditions and limiting the produc­
h, and 48 h and it was found that maximum yield of banana pseudostem tion of undesirable byproducts (Saritha et al., 2013; Sharma et al.,
(53.54% (w/w)) was obtained at 773 K with increased oxygen content. 2021). Additionally, enzymatic hydrolysis requires low maintenance
cost and generates higher sugar yield whereas acid hydrolysis needs a
4. Bioalcohol production from banana wastes better disposal system and it slowly degrades sugar monomer (Ferreira
et al., 2009). The cost of cellulase enzymes, which contributes to 30 to
Various types of banana waste have been explored as potential 50% of the overall cost of enzymatic saccharification of lignocellulose
sources for the bioethanol production through different conversion for ethanol generation, is the primary limitation. Legodi et al. (2021)

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A.N. Sawarkar et al. Bioresource Technology Reports 18 (2022) 101092

Cellulose

Lignin

Pretreatment Product
Hydrolysis Hydrolysate Fermentation
Recovery

Hemicellulose Bioethanol

Banana agro wastes

Fig. 2. Schematic representation of steps involved in biochemical conversion of banana wastes to bioethanol.

studied the crude enzyme from Trichoderma harzianum LMLBP07 13-5 sugars into simple chemicals and produces alcohol as a by-product.
and T. longibrachiatum LMLSAUL 14-1 for enzymatic hydrolysis of Various bacteria and yeast metabolizes monosaccharides (e.g., glucose
pretreated banana pseudostem. According to their investigation, and fructose) and disaccharides (e.g., maltose and sucrose), resulting in
saccharification of liquid hot water pretreated pseudostem released the generation of bioethanol from banana wastes. The efficiency of any
21.8 g/L of glucose at substrate loading of 15% and crude cellulase microorganism as a bioethanol fermenter can be determined based on its
loading of 10 filter paper unit (FPU)/g by Trichoderma harzianum proficiency under different process conditions such as osmotic tolerance,
LMLBP07 13-5. Saccharification by crude cellulase of T. long­ genetic stability, pH, temperature, feed concentration, residence time,
ibrachiatum LMLSAUL 14-1 released 38.6 g/L of glucose at substrate nutritional needs, etc. (Gupta and Verma, 2015; Kang and Lee, 2015). In
loading of 12.5% and cellulase loading of 10 FPU/g from banana general, the reliable bio agent for producing bioethanol should possess
pseudostem. John et al. (2020) investigated enzymatic hydrolysis of acid important characteristics, viz. capability to grow in low-cost and simple
assisted ultrasonication pretreated banana peel using Aspergillus niger. growing medium, high bioethanol yield (>90%, theoretical estimation),
They reported 9.3 g/L of maximum reducing sugar after 96 h of enzy­ high bioethanol tolerance (>40.0 g/L), tolerance to inhibitors in the
matic hydrolysis with 40 FPU of isolated cellulase at 50 ◦ C and 150 rpm. metabolism, and potential to retard the containments from growth
Further, cellulase activity of the isolated crude was found to be 1.712 media.
FPU/mL. Guerrero et al. (2018) investigated enzymatic hydrolysis for As per the literature, Saccharomyces cerevisiae is the most commonly
banana pseudostem and rachis. The researchers reported 75 g/L of utilized yeast for ethanol fermentation as it has been established to be
glucose conversion from banana pseudostem at 15.1% of solid loading robust and well adapted to the fermentation of banana wastes hydro­
and 14.9 FPU/g glucan. Further, they reported 96.0 g/L of glucose lysates. Saccharomyces cerevisiae uses the Embden-Meyerhof-Parnas
conversion for rachis at optimum parameters of 17.6% of solid loading (EMP) pathway to take in glucose molecules (majorily hexose) and
and 16.0 FPU/g glucan of enzyme dosage. Nwabanne and Aghadi (2018) generate ethanol. Li et al. (2021) studied the variation in biomass
employed Aspergillus niger for hydrolysis of banana peel and reported enzymatic saccharification and bioethanol production among 11
122 mg/mL of sugar yield at 34 ◦ C, pH 6.5, and for residence time of 5 different variations of banana pseudostems and rachis samples with
days. Suhag et al. (2020) investigated enzymatic hydrolysis of dilute chemical and hot water pretreatment. They stated that the banana crop
acid pretreated banana leaves by employing Aspergillus niger JD-11 in showed complete enzymatic saccharification which leads to bioethanol
incubator shaker under optimum conditions of pH 5.0, 150 rpm for 70 h. yield up to 31–38% (% dry matter) containing a high proportion of
The researchers attained highest amount of reducing sugars by crude edible carbohydrates (soluble sugar, starch) and lignocellulose. They
cellulase (524.83 mg/g) at 45 ◦ C, 15 FPU/g enzyme loading, and 2% also reported the role of two wall polymer cellulose CrI and lignin G-
substrate loading. Shimizu et al. (2018) investigated enzyamatic hy­ monomer in enzymatic hydrolysis of banana waste. Shankar et al.
drolysis utilizing 15 FPU/g total cellulase and 15 U/g glucosidase for (2020) reported Saccharomyces cerevisiae as a promising strain for pro­
alkaline, acidic and peroxide pretreated banana pseudostem. It was re­ ducing bioethanol from hexoses in banana leaves. Saccharomyces cer­
ported that the enzymatic hydrolysis of alkali pretreated pseudostem evisiae possesses better efficiency in sugars to alcohol conversion along
had the maximum glucose yield (85%), followed by acid pretreatment with tolerance to high ethanol concentrations for banana peels (Palacios
(82% glucose yield), and peroxide pretreatment (74% glucose). Guo et al., 2019). Oberoi et al. (2011) reported that Saccharomyces cerevisiae
et al. (2018) reported enzymatic hydrolysis of acidic pretreated banana yeast produced highest ethanol concentration with 28.2 g/L from ba­
peel with the highest conversion rate of 48.41% of reducing sugar, nana peels. Kluyveromyces marxianus, a thermotolerant yeast has also
implying that almost half of the dried peel powder by weight was hy­ been utilized for fermentation of banana residue hydrolysate. It can
drolyzed into fermentable sugars by cellulase. Tiwari et al. (2019) withstand high temperature which is necessary for enzymatic hydroly­
conducted enzymatic hydrolysis by employing enzyme from Aspergillus sis. Gatdula et al. (2021) and Palacios et al. (2017) explored Kluyver­
niger to microwave pretreated assisted banana peel. Maximum reducing omyces marxianus as a viable bio-agent capable of generating bioethanol
sugar of 0.561 g/g was reported under enzyme loading of 30 FPU/g, pH from banana pseudostem and banana peels, respectively. Ingale et al.
5 at 50 ◦ C and 150 rpm for 48 h. Under co-culture fermentation on (2014) explored a yeast strain S. cerevisiae NCIM 3570 for ethanol syn­
alkaline pretreated banana pseudostem, Ingale et al. (2014) employed thesis from banana pseudostem. In addition to these, attempts have also
two cellulase producing fungal strains, A. fumigatus and A. ellipticus, been made by researchers to investigate co-culture enzymes for the
which accounted for 41% saccharification with 833 mg% of reducing fermentation of banana wastes. In this line, Reddy et al. (2010) inves­
sugars at 5 g of substrate loading, pH 5 at 50 ◦ C for 48 h of incubation. tigated fermentation of banana leaves by coupling Clostridium thermo­
Kusmiyati and Sukmaningtyas (2018) obtained 29.8 g/L reducing sugar cellum CT2 and C. thermosaccharolyticum HG8 for bioethanol production.
content from alkali pretreated banana pseudostem by utilizing cellulase
enzyme for enzymatic hydrolysis. 4.1.3. Different fermentation schemes for bioethanol production from
banana wastes
4.1.2. Fermentation mechanism and role of microbes in banana wastes The design for fermentation of banana waste hydrolysate employs a
conversion special approach in which fermentation is carried out in multiple phases
Fermentation is a microbial oxidation process that converts soluble following cellulose hydrolysis. The most widely utilized schemes for

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A.N. Sawarkar et al. Bioresource Technology Reports 18 (2022) 101092

bioethanol synthesis from banana wastes includes separate hydrolysis pseudostem in an SSF scheme by employing Saccharomyces cerevisiae
and fermentation (SHF), simultaneous saccharification and fermenta­ Ethanol Red. They reported that SSF resulted into glucose concentration
tion (SSF), pre-saccharification and subsequently simultaneous of 27.5 g/L after a reaction time of 8 h. The release of glucose units and
saccharification and fermentation (PSSF) and continuous flow immediate consumption by yeasts are at the core of these activities. This
fermentation. impact resembled that there was no glucose left after 50 h. Enzymes, on
In SHF, hydrolysis and fermentation steps are operated sequentially. the other hand, continued to hydrolyze oligomers in the medium which
Herein, pretreated lignocellulosic biomass is first hydrolyzed to glucose in turn resulted in the rise of ethanol produced until 72 h. 46.2 g/L of
and then fermented to bioethanol, in separate reactors. The primary ethanol production with a maximum attainable yield of 82.8% was
advantage of SHF is that both stages are produced at optimal tempera­ achieved after 72 h of reaction time from rachis. They further reported
tures for cellulase enzymes and yeast. The disadvantage of the method is that SSF of banana pseudostem accounted for 40.1 g/L ethanol pro­
the buildup of hydrolysis products (mainly cellobiose and glucose duction with maximum attainable yield of 70.8% at 72 h. Kusmiyati and
sugars) in the enzymatic reactor, which produces feedback inhibition of Sukmaningtyas (2018) demonstrated SSF by employing cellulase
the cellulolytic enzyme system. Further, use of two separate reactors for enzyme and Saccharomyces cerevisiae to alkali pretreated banana pseu­
SHF process raises the investment expenses (Saha et al., 2011). Another dostem and reported 4.32 g/L ethanol production. Oberoi et al. (2011)
important issue with SHF is microbiological contamination as a result of explored SSF scheme by utilizing S. cerevisiae to hydrothermally pre­
the prolonged incubation period in hydrolysis. The enzymes could be treated banana peels for the production of bioethanol and reported that
possible source of contamination, and sterilizing them in a large-scale banana peels produced 28.2 g/L ethanol concentration and bioethanol
process remains challenging. In one of the recent studies, Legodi et al. productivity of 2.3 g/L/h.
(2021) investigated SHF of alkaline pretreated pseudostem hydrolysate In PSSF scheme, the enzymes are introduced to the pretreated
employing Saccharomyces cerevisiae UL01 at 30 ◦ C and 100 rpm. The biomass to begin the saccharification process, and a few hours later,
highest amount of ethanol produced was 17.6 g/L. Guerrero et al. (2018) when the glucose begins to accumulate, the fermentative microbe is
investigated SHF scheme for pretreated acid catalyzed steam explosion added to convert it to ethanol, preventing glucose buildup in the reactor.
of banana rachis and pseudostem. According to their findings, rachis As a result, during the initial hours of the process, the enzymes may
produced 48.0 g/L of bioethanol, with maximum attainable yield of function at their ideal temperature for glucose release, and the viscosity
85.9% after 120 h of the process (72 h involved for enzymatic hydrolysis in the system stays low. The technique sought to be cost effective as
and 48 h for the fermentation) using Saccharomyces cerevisiae. Similarly, hydrolysis and fermentation are carried in the single reactor. Further,
for 120 h of processing, the highest bioethanol content in the broth for PSSF is considered to be superior to SSF and SHF in terms of bioethanol
banana stem was found to be 42.0 g/L with a yield of 74.3%. Further, the concentration and bioethanol conversion. However, longer reaction
study concluded that the usage of SHF eliminates the filtration proced­ time during the conversion process is the major bottleneck associated
ure, thereby lowering producing costs. Uchôa et al. (2021) explored with PSSF. Guerrero et al. (2018) investigated the suitability of banana
integrated usage of three different banana residues composition in the rachis and pseudostem as a substrate for bioethanol production in a PSSF
fermentation broth. The wet mass ratio of between banana pulp, peels, scheme. According to their findings, acid catalyzed steam exploded
and pseudostem bagasse of 1:2:10 was utilized. According to the au­ banana rachis produced the maximum attainable bioethanol yield
thors, simultaneous fermentation of three waste banana residues by co- 86.6% in 72 h time of reaction for 17.6% of solid loading and 16.0 FPU/g
culture of S. cerevisiae and P. tannophilus achieved the highest produc­ of enzyme dosage. Alternatively, banana pseudostem attained the
tivity and yield in ethanol. maximum yield of bioethanol (73.9%) in 8 h reaction time under 15.1%
SSF scheme combines hydrolysis and fermentation in a single of solid loading and 14.9 FPU/g of enzyme dosage. Suhag et al. (2020)
reactor. In this method, the glucose produced by the hydrolyzing en­ investigated banana leaves as a potential substrate for bioethanol pro­
zymes is promptly consumed by the fermenting microbe present in the duction. According to their findings, after 12 h of enzymatic hydrolysate
culture, and a low concentration of sugars is kept in the media, thereby fermentation, the highest value of 15.43 g/L of ethanol was achieved,
minimizing the problem of cellulase end product inhibition (Hans et al., with a yield of 0.38 g/g sugar and a productivity of 1.28 g/L/h. Ingale
2019). The major advantages of SSF are high ethanol yield, less con­ et al. (2014) conducted fermentation of banana pseudostem by
version time, less sterile condition requirement, low enzyme require­ Saccharomyces cerevisiae NCIM 3570 and reported the ethanol yield of
ment, cost saving by eliminating expensive reactions and separation 84%. The substrate was dried overnight after pretreatment with acid,
equipment. However, inhibition of cellulase enzyme by ethanol gener­ alkali, and steam and treated with two sets of saccharification. It has
ated during fermentation is a primary drawback of SSF. Variation in the been found that alkali treatment with enzymatic saccharification (833
optimal temperatures for enzymes involved in hydrolysis, yeast, and ± 9.6 mg%) produced more reduced sugar than alkali treatment (203 ±
various microorganisms involved in the fermentation process is another 8.3 mg%). Palacios et al. (2019) produced ethanol from banana peels
primary concern (Rastogi and Shrivastava, 2017). The optimal tem­ using two yeasts through different process configurations. Saccharo­
perature for enzymatic hydrolysis is 40–50 ◦ C, however, microorgan­ myces cerevisiae and Kluyveromyces marxianus strains were used with
isms with high ethanol production and yield do not generally withstand simultaneous saccharification and fermentation (SSF) and pre-
this high temperature. Employing thermotolerant microorganisms, like saccharification and simultaneous saccharification and fermentation
Kluyveromyces marxianus, which is engineered to resist higher temper­ (PSSF) at high solid loading, to the extent of 25% w/w. S. cerevisiae
atures vital for enzymatic hydrolysis can overcome this drawback. strain with 25% (w/v) BP on PSSF process produced maximum ethanol
Another significant disadvantage of SSF is the partial hydrolysis of the (32.6 g/L) whereas S. cerevisiae and K. marxianus produced 13 g/L and
substrates at the end of the process, which leads the yeast and adsorbed 11 g/L at 35 and 41 ◦ C on SSF with 10% (w/w) BP and did not show any
cellulases to form a close association with the recalcitrant residue (Toor significant difference for ethanol production by SSF. It was further
et al., 2020). This limits the reuse of the high concentration of yeasts revealed that PSSF is superior to SSF as it results in higher yield of
required to achieve reasonably high ethanol generation in the subse­ ethanol and can be applied to high concentration of banana peel but it
quent batch. As a consequence, much of sugars produced during cellu­ requires more time compared to SSF. However, fermentation of sugars is
lose hydrolysis are utilized to nourish yeast rather than sugars to faster in S. cerevisiae compared to K. marxianus with equal or greater
ethanol. Gatdula et al. (2021) studied SSF scheme of chemically pre­ amount of ethanol. Saccharomyces cerevisiae is the widely used yeast for
treated banana pseudostem and reported 5.35 g/L ethanol by employing ethanol production. Guerrero et al. (2018) produced bioethanol from
Kluyveromyces marxianus at 35 ◦ C with a 24 h reaction time at the banana pseudostem and banana rachis through fermentation by
enzyme loading of 30 FPU/g. Guerrero et al. (2018) investigated the Saccharomyces cerevisiae. The maximum attainable yield of ethanol from
potential of pretreated acid catalyzed steam explosion banana rachis and banana pseudostem and rachis was found to be 74% and 87%,

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respectively. Shankar et al. (2020) studied the potential of banana leaves are also being explored, lately, for pre-treatment of banana wastes which
as a substrate for ethanol production. They initially pretreated the leaves are proving efficient in terms of producing less inhibitors, operation ease
with acid thereby extracting the cellulosic fraction and hemicellulosic and high heating capability in short time. In hydrolysis process, the
sugars. The hydrolysate produced by enzymatic saccharification of the major challenge is to procure efficient cellulase for depolymerisation of
cellulosic portion of banana leaves was fermented to ethanol by cellulose and hemicellulose for producing fermentable monomers in
Saccharomyces cerevisiae, yielding 8.1 g/L of ethanol. Souza et al. (2014) high concentration. In context to hydrolysis of banana residues, Asper­
investigated bioethanol production by utilizing banana pseudostem as a gillus niger is extensively utlised for hydrolysis. Further, there is a wide
fermentation substrate and reported 22.1 g/L of ethanol production scope to explore potent, low cost, cellulase enzyme alternatives for
from the fermentation. Matharasi et al. (2018) employed Saccharomyces effective saccharification of complex banana residue polymer.
cerevisiae (KX033583) strain to the hydrolysate obtained by enzymatic Fermentation of various types of banana waste leading to the pro­
hydrolysis of banana pseudostem and banana peel. They reported that duction of bioethanol is highly sensitive to substrate and product re­
banana pseudostem produced 15.61 ± 0.07 g/L of ethanol yield at covery. In general, continuous fermentation processes are found to be
35 ◦ C, pH of 6 and at 5% of inoculum levels. On the other hand, banana economical, have high solvent productivity and developed to reduce the
peels hydrolysate produced 17.00 ± 0.07 g/L of ethanol. Meena et al. cell washout problem with a disadvantage of contamination problem
(2015) separately treated banana pseudostem with two distinct cellulose (Kumar and Gayen, 2011). These problems can be addressed through the
degrading bacteria (A2 and A3). After pretreatment, the residue was available computational tools as well as developed biotechnological
subjected to fermentation by utilizing Saccharomyces cerevisiae and re­ approaches focusing on the system biology (Bhatia et al., 2012). Future
ported that higher yield (0.288 g/g) of ethanol from residue degraded studies can be concentrated on consolidated bioprocessing wherein
with A2. Conversely, pseudostem pretreated with A3 bacteria resulted in cellulase production, banana waste hydrolysis, and ethanol fermenta­
less ethanol yield (0.196 g/g). Arumugam and Manikandan (2011) tion are collectively carried in a single reactor. Clostridium phyto­
investigated that the hydrolysates produced from the dilute H2SO4 fermentans, a microbe that can effectively ferment cellulose straight to
pretreated banana fruit peels produced 13.84% ethanol with a fermen­ ethanol, may be employed in this method. Consolidated bioprocessing of
tation efficiency of 27.13% at 42 h of incubation. banana wastes may assist in overall economization of the process by
Apart from the above discussed fermentation schemes, Rosario and lowering the number of unit operations, resulting in considerable energy
Pamatong (1985) investigated continuous flow fermentation of banana and operational cost reductions (Dahman et al., 2019).
fruit pulp sugar into ethanol. Continuous fermentation includes the Optimization of various parameters in subsequent process is still
continuous and simultaneous addition of substrate and harvesting of end challenging which critically affects the bioethanol production. Optimi­
product. Cell immobilisation methods are frequently utilized in contin­ zation of pre-treatment parameters, enzyme concentrations, saccharifi­
uous flow fermentation, allowing for cell recovery and ongoing usage. cation and fermentation parameters with the aid of statistical design
However, continuous fermentation procedures are more difficult to could prove beneficial in terms of improving ethanol productivity from
carry out, especially when the processes are to be maintained for a long banana residues, thereby evincing commercial potential of the process.
duration of time. According to their findings, ethanol productivity of 15 In this line, Oberoi et al. (2011) and Gebregergs et al. (2016) studied the
gdm− 3 h− 1 and ethanol yield of 89% of theoretical was achieved by optimization of different parameters during the process and achieved
utilizing carrageenan immobilized yeast (S. cerevisiae) loading of 13.4 g/ significant bioethanol productivity from banana wastes. More studies in
l at dilution rate of 0.28 h− 1. Table 4 presents a summary of the reported this direction need to be conducted.
investigations including pretreatment techniques, conditions, conver­ Challenges also exist with respect to feedstock collection and supply.
sion processes, microorganism employed for bioethanol production Innovation in collecting and channelling a vast amount of banana wastes
from various banana wastes. is a continuous challenge as various banana wastes are produced from
the field of banana cultivation. Also, one of the major sources of banana
5. Challenges and plausible remedies waste i.e. banana peels is available in households. A comprehensive
policy as regards the collection of both these major banana wastes needs
. The management of various agricultural wastes poses many envi­ to be implemented. Participation of the private sector in collection,
ronmental and sustainability concerns (Russ and Pittrof, 2004). The segregation, supply, and storage services of various banana wastes to the
growth of pathogens, bacteria, microbial action and biological insta­ bioalcohol conversion set-up facility can serve as an effective approach
bility are the significant issues while collecting and the storage of the for the government to minimize its financial load. Cooperatives or other
banana biomass (Gupta et al., 2022). The conversion of agricultural local bodies might be incentivized to collect and distribute a set amount
biomass such as various banana wastes into value-added products like of banana agro residues over a long period of time.
ethanol will be attractive if the product satisfies market demands and
standards and the process of its production is cost effective (Jayathi­ 6. Research needs and future directions
lakan et al., 2012). In this context, it is important to optimize the process
conditions for improving the yield of ethanol while limiting the by- Fig. 3 depicts research needs and future directions for bioethanol
products generation in the process. There are several other challenges production from banana wastes. To take the subject matter forward,
that need to be encountered for economically viable bioethanol pro­ there is tremendous scope for biological research leading to further in­
duction from banana wastes. The most common bottlenecks confronted roads in banana plant breeding. Plant genetic engineering can be
include the aspects of the pre-treatment, hydrolysis process and extensively researched to exploit deconstruction of banana plant cell
fermentation configuration. With regard to pre-treatment, it can be wall polysaccharides, inhibition of lignin biosynthesis enzymes, and
ascertained from literature that acid and alkali pre-treatment are increase of polysaccharide levels in banana plants for potential
conventionally employed methods for rendering banana components enhanced bioethanol production.
(cellulose and hemicellulose) amenable for hydrolysis. However, both Fundamental studies to understand the effect of each pre-treatment
the pre-treatments involve various disadvantages. Application of technique on various banana wastes at a structural and molecular
organic acids such as oxalic, tartaric could prove beneficial in terms of level need to be carried out. Further research on the pretreatment con­
less production of inhibitors compounds, safe and corrosive free. ditions to enhance the release of sugar concentrations from saccharifi­
Exploring lime pre-treatment for banana wastes could be regarded as cation is of immense interest to upsurge the bioalcohol-banana wastes
alternative viable techniques as it is comparatively cheap than alkaline ratio prior to scale-up. The transport mechanism pertaining to the
agents. Further, lime can be recovered easily as calcium carbonate by various pretreatment technologies for banana wastes needs to be thor­
neutralising with carbon dioxide. Microwave and ultrasonic irradiation oughly investigated for determining the rate controlling mechanism/s

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Table 4
Pretreatment techniques, conditions, conversion processes, microorganisms employed for bioethanol production from various banana wastes.
Banana Pretreatment Pretreatment Conversion process Microbial organism Optimum Conversion/ Reference
waste technique condition fermentation yield
conditions

Leaves Alkali and acidic Aqueous solutions of Fermentation Saccharomyces cerevisiae Initial sugar 15.43 g/L (Suhag et al.,
pretreatment, 0.1 N NaOH and 0.1 concentration- 40 g/ 2020)
steam N H2SO4 (1:10 (w/ L, yeast loading-(5%
pretreatment v)) autoclaved at v/v), pH 5.5, 150
121 ◦ C for 1 h rpm, temp-30 ◦ C,
fermentation time-30
h
Acidic 2.5% (v/v) dilute Fermentation Saccharomyces cerevisiae 10% of inoculum 8.1 g/L (Shankar et al.,
H2SO4, autoclaved at S. cerevisiae 2020)
121 ◦ C, 30 min inoculated at temp.
30 ◦ C, 150 rpm, time-
96 h
Alkali and acidic 1% of NaOH or 1% Co-culture Clostridium thermocellum Inoculum size 5%, pH 22 g/L (Reddy et al.,
pretreatment H2SO4, autoclaved at fermentation (under CT2 7.5, temp-30 ◦ C, 2010)
121 ◦ C, 15 min anaerobic conditions) C. thermosaccharolyticum incubation period-5
HG8 days
Pseudostem Acidic 0.5% v/v of H2SO4 Simultaneous Kluyveromyces marxianus Enzyme loading of 30 5.35 g/L (Gatdula et al.,
incubated in shaker saccharification and FPU/g, temp-35 ◦ C, 2021)
at 40 ◦ C, 150 rpm for Fermentation (SSF) reaction time-24 h
1h
Alkaline 3% of NaOH Separate hydrolysis Saccharomyces cerevisiae Inoculation of 17.6 g/L (Legodi et al.,
autoclaved at 121 ◦ C, and fermentation hydrolysate medium 2021)
15 psi for 1 h (SHF) with microorganism
in incubation temp-
30 ◦ C, rpm-150, time-
48 h
Acid-catalyzed 177 ◦ C, 5 min, 2.2% Separate hydrolysis Saccharomyces cerevisiae Solid loading-15.1%, 42 g/L (Guerrero et al.,
steam explosion H2SO4 (v/v) and fermentation Ethanol Red enzyme dosage-14.9 2018)
(SHF) FPU/g
Alkali 2 N of NaOH Simultaneous Saccharomyces cerevisiae Enzyme dosage- 20 4.32 g/L (Kusmiyati and
pretreatment autoclaved at 121 ◦ C Saccharification and FPU/g, 10% (v/v) Sukmaningtyas,
for 90 min fermentation (SSF) yeast, temp-120 ◦ C, 2018)
time-120 h
Physical Grinded with electric Fermentation Saccharomyces cerevisiae Inoculum level-5%, 15.61 g/L (Matharasi et al.,
grinder and further pH -6, temp-35 ◦ C 2018)
sieved
Biological Pretreated with Fermentation Saccharomyces cerevisiae Incubation time-24 h, 14.4 g/L (Meena et al.,
degrading bacteria incubation temp- 2015)
A2 for 72 h 30 ◦ C at 100 rpm
Biological Pretreated with Fermentation Saccharomyces cerevisiae Incubation time-24 h, 9.8 g/L (Meena et al.,
degrading bacteria incubation temp- 2015)
A3 for 24 h 30 ◦ C at 100 rpm
Chemical 120 ◦ C/15 min Fermentation Saccharomyces cerevisiae Inoculum agitation 22.1 g/L (Souza et al.,
pretreatment H2SO4 (2% m/m) frequency-120/min, 2014)
NaOH (3% m/m) temp-◦ C, pH 4.5.
Alkali and 18 h Fermentation Saccharomyces cerevisiae Hydrolysate conc-4.1 17.1 g/L (Ingale et al.,
microbial NCIM 3570 g% sugar, yeast cells – 2014)
treatment 5% inoculation, pH-
5.5, incubation temp-
30 ◦ C, agitation time-
72 h
Peels Acidic with 2% (v/v) H2SO4, 60 Fermentation Saccharomyces cerevisiae Inoculation with 5% 4.24 g/L (John et al.,
ultrasonification min subjected to (v/v) of S. cerevisiae, 2020)
ultrasonification at temp-37 ◦ C, shaker
operating frequency speed-100 rpm.
of 20 kHz and
750 W
Autoclaving 121 ◦ C, 15 min Presaccharification Saccharomyces cerevisiae 25% (w/v) of banana 32.6 g/L (Palacios et al.,
and Simultaneous peel, yeast load-2 g/L, 2019)
Saccharification and pH-4.8, shaker speed-
Fermentation (PSSF) 150 rpm, temp-35 ◦ C,
time-64 h
Physical Grinded with electric Fermentation Saccharomyces cerevisiae Inoculum level-5%, 17.00 g/L (Matharasi et al.,
grinder and further pH -6, temp-35 ◦ C 2018)
sieved
Acidic 0.5% v/v H2SO4 acid Fermentation Kluyveromyces marxianus 20% (w/w) of 21 g/L (Palacios et al.,
concentration, hydrolysate banana 2017)
121 ◦ C, autoclaved peel, time-24 h, 6.5 ×
for 15 min 107 cells/mL, temp-
42 ◦ C, shaker speed-
150 rpm
Acidic Fermentation Saccharomyces cerevisiae 45 g/L
(continued on next page)

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A.N. Sawarkar et al. Bioresource Technology Reports 18 (2022) 101092

Table 4 (continued )
Banana Pretreatment Pretreatment Conversion process Microbial organism Optimum Conversion/ Reference
waste technique condition fermentation yield
conditions

1.50% v/v H2SO4 5 g/L yeast, temp- (Gebregergs

concentration, 91 ◦ C, 30 ◦ C, speed of et al., 2016)
21.66 min retention incubator shaker-200
time rpm, pH -5
Biological Pretreated with Fermentation Saccharomyces cerevisiae Incubation time-24 h, 4.75 g/L (Meena et al.,
degrading bacteria incubation temp- 2015)
A2 for 72 h 30 ◦ C at 100 rpm
Biological Pretreated with Fermentation Saccharomyces cerevisiae Incubation time-24 h, 4.65 g/L (Meena et al.,
degrading bacteria incubation temp- 2015)
A3 for 24 h 30 ◦ C at 100 rpm
Hydrothermal 10% (w/v) banana Simultaneous Saccharomyces cerevisiae Solid-to-liquid 28.2 g/L (Oberoi et al.,
sterilization peel and nutrient Saccharification and loading-10% (w/v), 2011)
autoclaved at 121 ◦ C, Fermentation (SSF) pH-5, agitation speed-
15 psi, 15 min 200 rpm, temp-50 ◦ C,
time-3 h, cell
concentration- 3 ×
109 cells/mL
Liquid hot water Banana peels Fermentation Saccharomyces cerevisiae pH-6, yeast loading-1 13.84 g/L (Arumugam and
followed by acidic slurried with g yeast/1000 mL, Manikandan,
treatment distilled water at Temp-35 ◦ C, 2011)
10% w/w autoclaved Incubation time-42 h
at 121 ◦ C for 15 min.
H2SO4 (0.05 N)
Rachis Acid-catalyzed 198 ◦ C, 5 min 1.5% Pre-hydrolysis and Saccharomyces cerevisiae Pre-saccharification 48.3 g/L (Guerrero et al.,
steam explosion H2SO4 (v/v) simultaneous Ethanol Red time-8 h, solid 2018)
saccharification and loading-17.6%,
fermentation (PSSF) enzyme dosage-16.0
FPU/g, reaction time-
72 h for PSSF

Enhancing polysaccharide levels in banana plants for high yield of bioethanol

Exploiting deconstruction of banana plant cell wall polysaccharides
Banana plant genetic Inhibition of lignin biosynthesis enzyme
Engineering

Study at molecular and structural level

Effect of pre-treatment Investigation of transport mechanism of pre-treatment to determine rate
technique controlling mechanism and improving efficiency of pre-treatment

Exploring potent, cheap, genetically modified microorganisms,

Enhancing hydrolysis and
multitasking strains, cocktail blends for effective hydrolysis and
fermentation
fermentation
Research
Needs Process engineering for
Well-balanced and ingenious combination of pre-treatment,
integrated approach hydrolysis, and fermentation with optimization of reduced energy,
chemicals and nutritional inputs
Optimization of various
parameters Optimization of pre-treatment parameters, enzyme concentrations, saccharification
and fermentation parameters with the aid of RSM and OA to achieve commercial
potential of bioethanol from banana residues
Advanced recovery Optimizing upstream and downstream operations for reducing operating costs
techniques

Exploring high yield separation and purification of bioethanol

Life Cycle Assessment

Accessing environmental impacts associated with all the stages of

life cycle of bioethanol from banana wastes

Fig. 3. Research needs and future directions for bioethanol production from banana wastes.

and hence to identify the efforts for improving the efficiency of treat­ which would enhance various aspects of fermentation such as yield,
ment methods. Moreover, clarity on prospective application of gener­ better and wider substrate use, and increased recovery rates. Other way
ated by-products needs to be established, which could be helpful in round, the emerging scope of the process growth may be investigated by
opting for a particular pretreatment method. developing multitasking strains capable of cellulase secretion along with
With reference to the hydrolysis of banana wastes, there is a scope for co-fermentation of glucose and xylose to enhance process cost
innovative techniques for depolymerizing cellulose and hemicellulose to effectiveness.
produce fermentable monomers with high concentrations. Besides, There is enormous potential for process engineering which would
research efforts in generating more efficient enzyme blends/cocktails for entail the use of integrated approaches for the improved yield of bio­
depolymerizing holocellulose polymers of banana wastes into ferment­ alcohol production from banana wastes (Sirohi et al., 2020). Process
able sugars moieties are needed. There exists a huge opportunity for the engineers' priority may be concentrated on a well-balanced and inge­
genetic engineering in developing genetically modified microorganisms nious combination of pretreatment, hydrolysis, and fermentation with

11
A.N. Sawarkar et al. Bioresource Technology Reports 18 (2022) 101092

genetically robust fermentation strain, synthetic hydrolyzing enzymes, Bhatia, S.K., Joo, H.-S., Yang, Y.-H., 2018. Biowaste-to-bioenergy using biological
methods – a mini-review. Energy Convers. Manag. 177, 640–660. https://doi.org/
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10.1002/ese3.910.1016/j.enconman.2018.09.090.
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