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PRBI-10195; No. of Pages 11 ARTICLE IN PRESS

Process Biochemistry xxx (2014) xxx–xxx

Contents lists available at ScienceDirect

Process Biochemistry
journal homepage: www.elsevier.com/locate/procbio

Review

Biotechnological production of xylitol from lignocellulosic wastes:

A review
Tiago Lima de Albuquerque a , Ivanildo José da Silva Jr. a , Gorete Ribeiro de Macedo b ,
Maria Valderez Ponte Rocha a,∗
a
Universidade Federal do Ceará, Departamento de Engenharia Química, Campus do Pici, Bloco 709, 60455-760 Fortaleza, CE, Brazil
b
Universidade Federal do Rio Grande do Norte, Departamento de Engenharia Química, Campus Universitário Lagoa Nova, CEP 59078-970 Natal, RN, Brazil

a r t i c l e i n f o a b s t r a c t

Article history: Several studies have sought to evaluate alternative routes for the production of xylitol, a polyol with
Received 20 May 2014 high employability in both the food and pharmaceutical industries. Large-scale production is typically
Received in revised form 25 June 2014 achieved by a chemical process of d-xylose hydrogenation. However, due to the conditions employed in
Accepted 15 July 2014
this procedure, it remains very costly. The biotechnological route has been the subject of intense research
Available online xxx
throughout the last decade because, apart from implying lower costs, many microorganisms can be used.
The employment of unconventional substrates as carbon sources for this production also stands out as a
Keywords:
very promising alternative. In addition to the possibility of being assimilated by many microorganisms,
Xylitol
Bioprocess
it becomes a good destination for the disposal of industrial waste. Thus, the aim of this review was to
Lignocellulosic wastes gather data on the biotechnological production of xylitol from agro-industrial waste, as well as to identify
Yeasts the best cultivation conditions (i.e. treatment of hydrolysate, pH, temperature and aeration) employed
Operating conditions in the process.
© 2014 Elsevier Ltd. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
2. Xylitol applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
3. Xylitol production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
3.1. Biotechnological production of xylitol from lignocellulosic waste . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
3.1.1. Waste utilization for xylitol production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
3.1.2. Microorganisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
3.2. Interferences obtained in the hydrolysis of lignocellulosic materials for xylitol production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
3.3. Growth conditions aimed at producing xylitol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
3.3.1. pH and temperature . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
3.3.2. Aeration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
3.3.3. Carbon sources . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
3.3.4. Nitrogen sources . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
3.3.5. Bioreactor operation modes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
3.4. Xylitol purification from fermented hemicellulosic hydrolysates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
4. Future prospects and conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00

∗ Corresponding author at: Departamento de Engenharia Química, Universidade Federal do Ceará, Campus do Pici, Bloco 709, Fortaleza, CE 60455-760, Brazil.
Tel.: +55 85 3366 9611; fax: +55 85 3366 9610.
E-mail addresses: valderez.rocha@ufc.br, valponterocha@yahoo.com.br (M.V.P. Rocha).

http://dx.doi.org/10.1016/j.procbio.2014.07.010
1359-5113/© 2014 Elsevier Ltd. All rights reserved.

Please cite this article in press as: Albuquerque TLd, et al. Biotechnological production of xylitol from lignocellulosic wastes: A review.
Process Biochem (2014), http://dx.doi.org/10.1016/j.procbio.2014.07.010
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1. Introduction price. The study of these researchers concluded that if a substantial

Xylitol (C5 H12 O5 ) is a polyol that has a hydroxyl group attached
to each carbon atom in its chain, and possesses a high sweetening
power, presenting 40% fewer calories than sucrose. It was discov-
ered in 1891 by chemist Emil Fischer (German, 1852–1919) and his
team, who obtained it in syrup form from the reaction of xylose
with sodium amalgam [1]. The main characteristic of this polyol
is its high cooling power inherent in its high endothermic heat
(34.8 cal g−1 ) [2]. It has been vastly cited in the literature for its
organoleptic characteristics and health benefits such as: high sol-
ubility [3], low glycaemic rates [4], lack of carcinogenicity [5] and
cariostatic properties [6]. Furthermore, xylitol does not participate
in Maillard reactions (which can cause the formation of browning
compounds in food) and thus does not reduce the nutritional value
of the proteins present [7]. These characteristics confirm its appli-
cability in food production processes, such as candies, caramels,
chocolates, ice creams, jellies, marmalades and beverages [7].
Industrial production of xylitol has been on-going for almost
four decades and, since the early years, solutions of purified d-
xylose typically undergo a catalytic hydrogenation process, under
conditions of high-temperature (80–140 ◦ C) and pressure (up to
50 atm) [8], until polyol formation is achieved (chemical process).
Due to the operating conditions and the need for purity in the
employed xylose, this traditional process becomes quite expensive.
Several studies have sought alternatives to chemical routes, with
particular attention paid to biotechnological processes, which led
them to become a major focus in scientific studies. Aiming to further
reduce processing costs and, in addition, searching for solutions to
the recycling of agro-industrial waste, the potential utilization of
renewable raw materials is being evaluated, such as bamboo [9],
vegetable residues [10] and sunflower stalks [11]. These wastes, in
turn, can be hydrolysed, generally by diluted acids, with further
release of d-xylose available for microbial xylitol conversion.
Xylitol production is expanding in a global level [12] and it is
currently estimated a strong demand for this product in the global
market, of more than 125,000 tons per year, with a value that is
relatively high ($4.50–5.50 per kg for bulk purchase by pharma-
ceutics and food companies and $20.00 per kg in supermarkets)
and therefore, encouraging the search for production processes
with lower costs. The industrial production of the substance is
performed chemically by a catalytic hydrogenation of the purified
xylose. This method utilizes specialized and expensive equipment
to achieve the high pressure and temperature conditions necessary
for the process [12–15]. Xylitol has a 12% share of the total polyols
market with rapid growth worldwide due to an increasingly health
conscious consumer and fast increase in chewing gum sales [14,15],
and it is approved for food use in over 50 countries [15].
Studies that aim exclusively on economic evaluation of xylitol
production by bioprocesses routes are not found in the literature, as
well as studies for comparing its costs to the chemical process. It is
known that renewable carbon sources, such as industrial residues,
can be converted into substances with higher added value, both
chemically and by biotechnological routes, and the choice of the
process should be based on a detailed assessment of sustainability,
resource availability and transport logistics [14].
Franceschin et al. [15] assessed the bioconversion of rye straw
into ethanol and xylitol, evaluating this integrated process econom-
ically. In that study, it was concluded that the capital required for
the execution of the bioprocess reached a total amount of about $40
million, with 25.9% and 29.0% of this amount approximately corre-
sponding to the costs with the fermentation and recovery of xylitol,
respectively. It is noteworthy that the economic analysis is strictly
dependent on the marketing price of xylitol, and it was observed
that the market would only be saturated with xylitol after a long
period, which would occasionally lead to a decrease in its selling
portion of hemicellulose used is converted into xylitol, this product
would generate all the required income to maintain the process.
Furthermore, in addition to the use of lignocellulosic materi-
als for the production of xylitol and ethanol, some studies [16–18]
also show the use of sugars obtained from such materials for the
production of polyunsaturated fatty acids with medicinal interest.
Thus, the aim of this article was to review alternative processes
for the biotechnological production of xylitol from agro-industrial
waste. The diversity of microorganisms used in the process was
studied, as well as the optimum conditions for polyol production.
2. Xylitol applications
In the European Economic Community the safety of xylitol use
has been recognized since 1984, and the agency that regulates food
and drugs in the United States (Food and Drug Administration –
FDA) has classified it as “Generally Recognized as Safe” (GRAS)
since 1986 and “Safe for Teeth” since 1994. Xylitol is extremely
well tolerated by the human organism when taken in doses over a
certain time frame with a maximum amount of 20 g, with the total
amount consumed not exceeding 60 g per day, since the intake of
higher doses presents laxative effects [2]. The literature includes
several studies on the use of xylitol as a replacement for sucrose
in certain foods aimed at caloric reduction, such as in cupcakes
[19], cookies [20], chocolate [21] and chewing gums [22]. The use
of xylitol in the pharmaceutical industry is now widespread, mainly
due to its proven anticariogenic properties, which can reduce tooth
decay by up to 100%. Several studies have shown the effective-
ness of this sugar-alcohol against the formation of oral biofilms,
especially when confronting the bacterium Streptococcus mutans
[23,24]. Some authors claim that the cariogenic characteristics of
xylitol against this bacterium are due to a decrease in glucosyl-
transferase expression in the microorganism [23]. Other authors
state that it changes the cell structure by turning the polysaccharide
that surrounds them into a diffuse and sparse cell wall [24]. Apart
from S. mutans, other studies have shown the effectiveness of xylitol
employment against other microorganisms that are harmful to oral
health, such as: Streptococcus pneumoniae and Haemophilus influen-
zae [24], Staphylococcus aureus [25] and Pseudomonas aerugionosa
[26]. Besides fighting tooth decay, xylitol is recognized for being
able to contribute to tooth calcification, which, in most cases, is
caused by ageing [27]. It has also been reported to be effective in the
treatment of several diseases, for instance: diabetes [4], anaemia
[28], acute otitis media [29] and osteoporosis [30].
3. Xylitol production
Xylitol is an intermediate product formed during carbohydrate
metabolism in mammals, including man, with a human being under
normal conditions able to produce 5–15 g of xylitol per day [31].
The commercial value of xylitol restricts its employment to cer-
tain foods and medicines, and due to this fact, new production
processes need to be devised in order to cheapen the costs inher-
ent in it [8]. Xylitol production may be performed by chemical
means, by xylose-hydrogenation reactions, or through microbio-
logical routes, in which specific microorganisms convert xylose to
xylitol. Apart from these, the enzymatic production process has
raised the interest of scientists given the fact that it is a biotechno-
logical alternative that can convert up to 100% of xylose to xylitol,
since no metabolic diversions for cell maintenance occur [32,33].
Industrial-scale production first started in 1975 in Finland, by
the Finnish Sugar Co. Ltda., in Helsinki. This process, which utilizes
chemical routes, is carried out in high-pressure (31–40 atm) and
high-temperature (100–130 ◦ C) reactors and lasts from three to 5 h,

Please cite this article in press as: Albuquerque TLd, et al. Biotechnological production of xylitol from lignocellulosic wastes: A review.
Process Biochem (2014), http://dx.doi.org/10.1016/j.procbio.2014.07.010
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Fig. 1. Xylitol production by chemical route.
depending on the employed conditions. The currently used large-
scale production is divided into several steps (Fig. 1): Step 1. Acid
hydrolysis of the xylan-rich natural material; Step 2. Hydrolysate
purification up to the point where a solution of pure xylose is
obtained; Step 3. Catalytic hydrogenation of pure xylose to xyli-
tol with a catalyst (a Ni-Al2 alloy); Step 4. Purification of the xylitol
solution obtained; Step 5. Xylitol crystallization [34,35].
The high temperatures and pressures employed required spe-
cialized and expensive equipment to achieve conditions necessary
for the process [12–15], not to mention the numerous purification
steps required, render the process of obtaining xylitol expensive
[12,13]; therefore, new means of production that can be as effec-
tive as many of the traditional methods have been sought [36].
Currently, Danisco (industry of DuPont Company) is a major global
supplier of xylitol, mainly from China, which it produces via catal-
ysis using xylose obtained from hardwood sources [15]. They have
recently developed an integrated process with the pulp and paper
industry. However conversion of xylose to xylitol is still achieved
via a chemical route.
3.1. Biotechnological production of xylitol from lignocellulosic
waste
3.1.1. Waste utilization for xylitol production
Xylose, along with glucose, is a major sugar derived from the
hydrolysis of lignocellulosic biomass, among other sugars such as
mannose, galactose, arabinose and rhamnose (all with countless
applications in both pharmaceutical and food industry) [37,38].
This substance is the second most abundant sugar found in nature,
Please cite this article in press as: Albuquerque TLd, et al. Biotechnological production of xylitol from lignocellulosic wastes: A review.
Process Biochem (2014), http://dx.doi.org/10.1016/j.procbio.2014.07.010
3
Fig. 2. Diagram of process for bioproduction of xylitol from lignocellulosic material.
and the process of converting xylose to xylitol has a signifi-
cant economic role regarding biomass employment [39]. Thus,
the development and optimization of methods both for obtain-
ing xylose from lignocellulosic wastes and for then converting this
sugar into products with higher added value, such as xylitol, is
of great interest. Fig. 2 shows a simplified flowchart of the steps
performed to produce xylitol using lignocellulosic materials.
The employment of waste generated by agribusiness has been
widely investigated for potential use of its raw materials in xyli-
tol production. It is known that these materials, after undergoing a
pretreatment, can release fermentable sugars such as d-xylose, the
main carbon source for xylitol production. Table 1 shows the variety
of waste that has been evaluated throughout the last decade for this
process. A pretreatment or hydrolysis step is performed initially
on these raw materials to expose their constitutive sugars so they
can be used later as a carbon source by different microorganisms,
enabling these raw materials to be susceptible to biotechnological
usage. Many of the pretreatments or hydrolysis steps that release
xylose from the hemicellulosic structure employ acids at dilute
concentrations, such as phosphoric acid [11] and sulphuric acid
[38,40]. The waste is then subjected to these acidic solutions at
temperatures generally above 100 ◦ C.
It is noteworthy that during the hydrolysis of lignocellulosic
materials, many compounds that can inhibit microorganisms can
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Please cite this article in press as: Albuquerque TLd, et al. Biotechnological production of xylitol from lignocellulosic wastes: A review.

Table 1
Xylitol production by different yeast strains and operational conditions using lignocellulosic waste as feedstock.

Microorganism Waste Culture conditions Maximum xylitol Fermentation mode Yield (g g−1 ) Productivity Reference
production (g L−1 ) (g L−1 h−1 )

pH Temperature (◦ C) Time (h)

T.L.d. Albuquerque et al. / Process Biochemistry xxx (2014) xxx–xxx

ARTICLE IN PRESS
Candida athensensis SB18 Vegetable waste 7.0 30.0 102 100.1 Batch mode (bioreactor) 0.81 0.98 [10]
Candida guilliermondii Sugar cane bagasse 5.5 30.0 120 50.5 Batch mode (Erlenmeyer flasks) 0.81 0.60 [37]
FTI20037
Candida guilliermondii Rice straw – 30.0 116 66.1 Batch mode (bioreactor) 0.84 0.17 [71]
FTI20037
Candida magnoliae Bamboo culm – 30.0 30 10.5 Batch mode (bioreactor) 0.59 0.42 [9]
Candida tropicalis As 2.1776 Corncob 5.5 30.0 120 96.5 Fed-batch mode (bioreactor) 0.83 1.01 [43]
Candida tropicalis BCRC 20520 Wood sawdust 5.0 30.0 96 41.4 Batch mode (Erlenmeyer flasks) 0.70 0.43 [77]
Candida tropicalis CCTCC Corncob 6.0 35.0 14 38.8 Fed-batch mode (bioreactor) 0.70 0.46 [36]
M2012462
Candida tropicalis HDY-02 Corncob 7.0 35.0. 78 58 Fed-batch mode (bioreactor) 0.73 0.74 [78]
Candida tropicalis JH030 Rice straw 6.0 30.0 80 31.1 Batch mode (Erlenmeyer flasks) 0.71 0.44 [90]
Candida tropicalis NBRC 0618 Olive pruning waste 5.0 30.0 25 53 Batch mode (bioreactor) 0.49 – [40]
Candida tropicalis W103 Corncobs 6.0 35.0 70 68.4 Fed-batch mode (bioreactor) 0.70 0.95 [91]
Debaryomyces hansenii Sugar cane bagasse 6.0 40.0 156 71.2 Batch mode (immobilized cells) 0.82 0.46 [39]
Debaryomyces hansenii CCMI Grain brewery 5.5 30.0 72 24.0 Batch mode (Erlenmeyer flasks) 0.57 0.51 [88]
941
Debaryomyces hansenii NRRL Vine waste 6.0 30.0 116 27.5 Batch mode (Erlenmeyer flasks) 0.54 – [89]
Y-7426
Hansenula polymorpha ATCC Sunflower stalks 5.5 30.0 169 0.31 Batch mode (bioreactor) 0.0023 – [11]
34438
Kluyveromyces marxianus CCA Cahew apple bagasse 6.0 30.0 96 6.76 Batch mode (Erlenmeyer flasks) – – [38]
510
Kluyveromyces marxianus CE Cashew apple bagasse 4.5 40.0 72 4.8 Batch mode (Erlenmeyer flasks) – – [87]
025
Pichia stipitis NRRL Y-30785 Corn stover 5.6 30.0 72 12.5 Batch mode (Erlenmeyer flasks) 0.61 0.18 [76]
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be formed. The main such compounds generated are furfural,
hydroxymethylfurfural, aliphatic acids, phenolic compounds and
vanillin [41–43]. Inhibitors present or formed from hydrolysates
can limit consumption of the carbon source and may even stop the
fermentation process from happening [41].
In many bioprocesses some treatment of the hydrolysate is
mandatory in order to eliminate these fermentative interferences.
This is mainly due to the fact that even in the smallest amounts
some of them can prevent microbial growth. Several detoxifica-
tion methods are used to remove toxic compounds from plant
biomass hydrolysates, such as enzyme biological treatments (lac-
case), physical treatments, evaporation and extraction, chemical
treatments with alkaline agents (NaOH, CaO, Ca(OH)2 ) and physico-
chemical treatments involving adsorption with activated carbon
and ion-exchange resins. Activated carbon [39] and ion-exchange
resins [37] have been used quite successfully to decrease or even
completely eliminate the major fermentation inhibitors. Addition-
ally, treatments employing microorganisms for detoxification of
lignocellulosic hydrolysates have received considerable attention
in recent years. Fonseca et al. [42] studied biological detoxifi-
cation of different hemicellulosic hydrolysates (corn fibre, straw
and sugarcane bagasse, coffee bean husks) using yeast Issatchenkia
occidentalis CCTCC M 206097. The greatest clearance of inhibitors
together with the lowest loss of reducing sugars was observed
when the hydrolysate was concentrated fivefold before the bio-
logical treatment. The total reductions observed within 24 h of the
experiment were: syringaldehyde (66.67%), ferulic acid (73.33%),
furfural (62%) and 5-HMF (82%). Mussatto and Roberto [2] treated
rice straw hydrolysate with five types of activated carbon and
observed that with carbons presenting lower particle sizes, greater
reductions in toxic compounds could be achieved.
3.1.2. Microorganisms
Different species of microorganisms, as well as several alter-
native raw materials, have been evaluated for the production of
xylitol by biotechnological methods, but most studies are restricted
to certain species of Candida. The biotechnological production of
xylitol, being less expensive, has the potential to be an advanta-
geous alternative to the chemical process that is conventionally
employed. Various types of microorganisms, including bacteria
and filamentous fungi, can be used in the process. Rangaswamy
and Agblevor [44], for example, selected 17 bacterial cultures of
the genera Serratia, Cellulomonas and Corynebacterium for poten-
tial xylitol production (with trials carried for 48 h at 30 ◦ C and
130 rpm in Erlenmeyer flasks), and found out that Corynebac-
terium sp. B-4247 was the strain with the highest production rates
(10.05 g L−1 ). Cirino et al. [45] described the production of xyli-
tol using Escherichia coli W3110, a genetically modified strain, and
achieved a production of up to 38 g L−1 of xylitol (30 ◦ C, 250 rpm
for 80 h). Other genetically modified bacteria are also promising
for xylitol production, such as Bacillus subtilis [46] and Corynebac-
terium glutamicum [47]. Studies with filamentous fungi for xylitol
production are scarce, but some researchers have reported positive
results with Hypocrea jecorina [48] and Trichoderma reesei [49]. In
the latter case, the authors studied a genetically modified strain of T.
reesei to produce xylitol from barley straw pre-treated with organic
solvents, with the process being conducted for 168 h at 30 ◦ C.
Iverson et al. [50] evaluated the increasing catabolic reducing
power output (NADH) of glucose catabolism for reduction of xylose
to xylitol by genetically engineered E. coli AI05. The obtained strain
could in theory simultaneously uptake glucose and xylose, and util-
izes glucose as a source of reducing power for the reduction of
xylose to xylitol, under anaerobic conditions.
Yeasts are still the primary focus of studies that aim to produce
xylitol from hemicellulosic hydrolysates and different species are
cited in Table 1. These microorganisms can produce xylitol as an
Please cite this article in press as: Albuquerque TLd, et al. Biotechnological production of xylitol from lignocellulosic wastes: A review.
Process Biochem (2014), http://dx.doi.org/10.1016/j.procbio.2014.07.010
5
intermediate metabolite during the metabolism of d-xylose. Xylose
reductase (XR) is a typical NADPH-dependent enzyme, whereas
xylitol dehydrogenase (XDH) requires NADP+. Thus, conversion of
d-xylitol occurs in two steps, a reduction step followed by an oxi-
dation step. d-Xylose is first reduced to d-xylitol by NADPH and
subsequently this metabolite is oxidized to d-xylulose by NADP+
(Fig. 3), with these two reactions considered limiting for d-xylose
fermentation and d-xylitol production [10,51].
Numerous microbial species have a metabolic system with
NADPH-dependent XR and NAD+-dependent XDH as cofactors and
these enzymes are induced by xylose [52]. Among the genera capa-
ble of producing xylitol, Candida is the most studied and stands as
a promising microorganism. With this type of microorganism, oxy-
gen availability is the most important factor in xylitol production
from d-xylose. Under limited oxygen conditions, oxidative phos-
phorylation cannot re-oxidize all the NADH generated. Thus, the
intracellular NADH concentration increases and leads to xylitol
accumulation.
Yablochkova et al. [53] studied the activities of XR and XDH in 11
different species of yeast (including Candida, Kluyveromyces, Pichia,
Torulopsis and Pachysolen) and found that Candida tropicalis Y-456
had the highest specific activity of XR: 6.57 ␮mol min−1 mgprotein −1 .
According to literature reports, microorganisms with high XR activ-
ity and NADPH-dependent are potential producers of xylitol from
d-xylose.
Misra et al. [54] also investigated xylitol production by 18 strains
of yeasts and, coincidentally, the Candida yeasts showed better pro-
duction performance. A strain of C. tropicalis achieved production
of 12.11 g L−1 of xylitol using 50 g L−1 initial xylose concentration
at 30 ◦ C, pH 5 and 200 rpm for 72 h.
Studies have shown variable values for xylitol production by
biotechnological processes using yeasts from industrial residues
(Table 1). It can be readily observed that the results are very
discrepant and are related to the different microbial species and
growth conditions involved (carbon and nitrogen sources, pH, aer-
ation, and so forth). Controlling these conditions is therefore of
fundamental importance for optimizing the xylitol production pro-
cess.
Zikou et al. [17] employed a Thamnidium elegans strain that pro-
duced a maximum xylitol quantity of 31 g L−1 during growth on
blends on xylose and glucose.
Zhang et al. [10] studied xylitol production using the yeast Can-
dida athensensis SB18 in agitated flasks and bioreactor performed
in both batch and fed-batch systems. The concentration, produc-
tivity and yield of xylitol obtained in this study were 100 g L−1 ,
0.81 g g−1 and 0.98 g L−1 h−1 , respectively, using hydrolysed hor-
ticultural waste containing 200 g L−1 xylose.
Villarreal et al. [55] evaluated the xylitol production from euca-
lyptus hemicellulosic hydrolyzate by Candida guilliermondii and,
under the best employed operating conditions, 32.7 g L−1 of xyli-
tol were produced after 48 h fermentation, which correspond to
0.68 g L−1 h−1 volumetric productivity.
In a study done by Rocha et al. [38], Kluyveromyces marxianus
CCA 510 produced ethanol and xylitol, using cashew apple bagasse
hydrolysate as culture medium and it reaching a concentration of
11.89 g L−1 and 6.76 g L−1 , respectively, at 96 h. More study exam-
ples related will be cited in the following topics.
3.2. Interferences obtained in the hydrolysis of lignocellulosic
materials for xylitol production
Lignocellulosic materials, including cellulose, hemicellulose and
lignin, are the most abundant materials on Earth. Hemicellulose is
the second most common component in plant cell walls (cellulose
being the first) and is a potential substrate for ethanol and xylitol
production [56]. Hemicellulose degradation by chemical processes
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Fig. 3. d-Xylose metabolic pathway in yeasts.
yields xylose as the main fraction, as well as variable fractions of
arabinose, mannose, galactose and glucose. Apart from these afore-
mentioned sugars, chemical hydrolysis of lignocellulose can release
35 other byproducts that act as microbial inhibitors [57].
Inhibitors can be divided into furan derivatives, such as fur-
fural and 5-hydroxymethylfurfural (HMF), phenolic compounds,
weak acids (levulinic and formic acid) and ions of heavy metals
(nickel, aluminium, chromium, etc.) [58]. Furfural and HMF are two
furan derivatives formed by the hydrolysis of sugars, hexoses and
pentoses, respectively. These furans are present at relatively high
concentrations in the hydrolysate and can act as potential microor-
ganism inhibitors. Furans are probably the most important group
of inhibitors, since the fermentability of hydrolysates obtained by
acidification is inversely related to the concentration of these com-
pounds. In addition, phenolic compounds cause loss of integrity in
biological membranes, affecting their ability to serve as selective
barriers and enzyme matrices [59]. Acetic acid, the main aliphatic
compound present, is released from acetyl-hemicellulose groups.
In turn, formic acid is a product of HMF degradation.
Several methods to detoxify the culture media can be employed,
such as physical (detoxification mediated by evaporation), chem-
ical (addition of Ca(OH)2 ), adsorption (using ion-exchange resins
and activated carbons) and biological (microbial and enzymatic) to
fight inhibitors of xylitol production [56].
Mateo et al. [60] studied several methods to detoxify acid
hydrolysates of olive pruning residues (liming with NaOH, CaO and
Ca(OH)2 and adsorption with activated carbon). When NaOH was
used, the decrease in phenolic compounds was minimal (28–31%)
for two different alkaline detoxification processes, and a slight
reduction in acetic acid (<7%) was achieved, although no sig-
nificant sugar loss was observed. The calcium-based treatments,
though, had a notable effect, removing more lignin-derived com-
pounds (56–71%), acetic acid (about 50%) and total furan content
(approximately 70%). However, a high loss of sugar also occurred,
Please cite this article in press as: Albuquerque TLd, et al. Biotechnological production of xylitol from lignocellulosic wastes: A review.
Process Biochem (2014), http://dx.doi.org/10.1016/j.procbio.2014.07.010
making this method unsuitable for pretreatment of olive pruning
hydrolysates. As for the use of activated carbon in this hydrolysate,
it was shown to be an excellent alternative to reduce inhibitors (46%
of acetic acid, 81% of phenolic compounds and 98% of total furans),
with the possibility of regenerating the carbon employed to reduce
the cost of the process.
3.3. Growth conditions aimed at producing xylitol
Several factors may influence microbial growth and, in order to
optimizing these, conditions should be adjusted according to the
specificity of each microorganism. The most extensively studied
factors in microbial growth that show some influence in biopro-
cesses are temperature, pH, aeration rate, inoculum concentration
and nutrients (carbon, nitrogen, and vitamin sources, etc.). Proper
control of these variables is of great importance for good per-
formance in obtaining high-quality products with high efficiency.
There have been many studies on the optimal environmental con-
ditions for yeast growth aimed at biotechnological production of
xylitol, and these are discussed below.
3.3.1. pH and temperature
Most studies employing yeasts in biotechnological processes
have been carried out by adjusting the temperature in the range
of 30–37 ◦ C, which proves to be optimal for these microorganisms
in xylitol production [36,61].
In some cases, however, higher temperatures may be employed;
for instance, the thermotolerant characteristic of the yeast K.
marxianus makes it stand out in fermentation studies when com-
pared to other microorganisms. This is because, in addition to its
enzymatic reactions being accelerated, the risk of contamination
by other microorganisms is diminished [62]. Rodrussamee et al.
[63] studied the potential of K. marxianus DMKU3-1042 for xyli-
tol and ethanol production from sugars present in hemicellulose
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hydrolysates at high temperatures (30 ◦ C, 40 ◦ C and 45 ◦ C). Cell
growth and sugar consumption were observed at all temperatures
studied, confirming that the strain can grow at high temperatures.
It is noteworthy that cell growth showed diminished intensity
as the temperature increased, though without ceasing. At 30 ◦ C,
ethanol production was 2.5 g L−1 at 72 h, and xylitol production
was 4.3 g L−1 after 48 h. Xylitol production was actually favoured
at 40 ◦ C obtained a concentration of 7.0 g L−1 .
Three strains of K. marxianus (IMB2, IMB3 and IMB4), isolated
from an Indian distillery were characterized for their growth in
xylose at high temperatures (40–45 ◦ C) and different pH values (4.5,
5.0 and 5.5). It was observed that the lower temperature (40 ◦ C) at
pH 5.5 was better for ethanol and xylitol yields, where values of
2.08 g L−1 and 7.36 g L−1 , respectively, were reached using the IMB4
strain after 96 h of cultivation [64]. In turn, the optimal tempera-
ture for Kluyveromyces sp. IIPE453 growth was at 50 ◦ C and pH 5.0,
reaching a maximal production of ethanol and xylitol of 82 g L−1
and 11.5 g L−1 at 48 h, respectively [65].
Srivani and Setty [66] attempted to find the optimal environ-
mental conditions for xylitol production by Candida parapsilosis
NCIM-3323, studying variations in temperature (25–35 ◦ C) and ini-
tial pH (3–6). Over the range of initial pH studied, it was observed
that xylitol productivity was maximal when the initial pH was
3.5. Above this, there was a decrease in production. At the tem-
perature ranges, it was further observed that the productivity of
xylitol increased until the temperature reached 30 ◦ C and, from
this temperature on, a drop in production occurred. Ramesh et al.
[67] investigated xylitol production from corncob hemicellulosic
hydrolysate by Debaryomyces hansenii var. hansenii (MTTC 3034)
using a statistical optimization of response surface approach. In
this study, the optimum temperature and pH for production of the
polyol were 31.8 ◦ C and 7.25 g L−1 , respectively.
With C. tropicalis, several studies have considered temperatures
of 30–35 ◦ C for xylitol production. Ping et al. [36], for example,
utilized C. tropicalis CCTCC M2012462 for xylitol production from
hydrolysed corncob at 35 ◦ C, reaching 38.8 g L−1 after 84 h of bio-
processing. Misra et al. [61] also used a corncob hydrolysate to
obtain 11.89 g L−1 xylitol at 30 ◦ C using a strain of C. tropicalis. El-
Baz et al. [68] studied the production of xylitol in synthetic media
at 30 ◦ C for C. tropicalis, obtaining a maximum concentration of
36.25 g L−1 .
3.3.2. Aeration
Yeasts do not form a homogeneous group regarding their energy
efficiency. The destination of pyruvate, a glycolytic intermediate,
determines the type of metabolism. When all the pyruvate pro-
duced is converted via the tricarboxylic acid cycle (respiration),
oxidative metabolism occurs and, if it is reduced to ethanol or other
compounds, oxy-reductive metabolism (fermentation) happens,
with the two pathways also able to occur simultaneously. Aeration
is seen as a major factor in the experimental production of xylitol,
and the amount of oxygen available is one of the determining fac-
tors for which pathway xylose will be shunted to, fermentation or
respiration, in turn regulating the carbon consumption balance for
both growth and bioconversion [68].
It is evident that yeasts require little oxygenation in the cul-
ture medium for d-xylose reduction and have specific rates for each
species [68].
To use and metabolize d-xylose, it must be converted to xylu-
lose. Xylulose is then phosphorylated to xylulose 5-phosphate by
xylulokinase (XK), with the latter being metabolized by the pentose
phosphate pathway. In bacteria, d-xylose is converted directly to d-
xylulose by xylose isomerase (XI). In eukaryotes such as yeasts and
filamentous fungi, d-xylose is converted to d-xylulose by means of
reduction and oxidation, which are catalyzed by xylose reductase
(XR) and xylitol dehydrogenase (XDH). The main limiting factor in
Please cite this article in press as: Albuquerque TLd, et al. Biotechnological production of xylitol from lignocellulosic wastes: A review.
Process Biochem (2014), http://dx.doi.org/10.1016/j.procbio.2014.07.010
7
this process is redox imbalance, which is caused by the difference
in the enzymes’ preference for coenzymes, NADPH in the case of XR
and NAD+ in the case of XDH. This redox imbalance is the key rea-
son for the activity by which the yeast accumulates xylitol during
anaerobic cultivation [69]. Fig. 3 shows the key reactions in d-xylose
metabolism during xylitol production using xylose as substrate.
Sirisansaneeyakul et al. [70] studied xylitol production by Candida
magnoliae TISTR 5663 in a fed batch and under oxygen-limited
conditions (1 vvm). Xylitol yields of 0.727 g g−1 , 0.719 g g−1 and
0.720 g g−1 were obtained with average biomass concentrations
of 21 g L−1 , 48 g L−1 and 50 g L−1 , respectively, under the evaluated
conditions (stirring speed 300 rpm, pH 7.0 and 30 ◦ C).
El-Baz et al. [68], in turn, evaluated the potential of C. tropicalis
in a synthetic medium containing 20 g L−1 xylose as carbon source
and concluded that xylitol production was increased when medium
oxygenation was reduced (by doubling the volume of medium in
the 250 mL Erlenmeyer flask from 20 mL to 40 mL). Mussatto and
Roberto [71] investigated xylitol production by C. guilliermondii FTI
20037 in a bioreactor from rice straw hemicellulosic hydrolysate
at different agitation rates (200 rpm, 300 rpm and 500 rpm). These
researchers concluded that xylose bioconversion to xylitol by the
yeast depends on the stirring speed, reaching a maximum yield of
0.84 g g−1 at 300 rpm.
3.3.3. Carbon sources
The use of lignocellulosic materials for the production of
high-added value bioproducts is promising, as they represent an
abundant and renewable supply of carbon sources. Lignocellulosic
materials such as corncobs, rice straw, sugarcane bagasse, cashew
bagasse, sawdust and oat hulls, among others (Table 1), represent
an important and cheap source of microbial substrates. The hemi-
cellulosic fraction can be hydrolysed to xylose and then fermented
to xylitol [9].
With respect to xylose utilization inside the cells, this compound
can be either metabolized through the phosphoketolase reaction,
which is the most efficient pathway yielding around 1.2 mole of
acetyl-CoA per 100 g of xylose (∼0.66 mole) utilized, or the pentose
phosphate pathway, where around 1.0 mole of acetyl-CoA is formed
per 100 g of xylose utilized [72].
Several studies have evaluated the use of corncob hydrolysate
as a carbon source for xylitol production [36,61,64]. Kamat et al.
[73] isolated a yeast strain, Cyberlindnera (Williopsis) saturnus, from
mangrove forests that was capable of producing xylitol (29.1 g L−1 )
from detoxified corncob hydrolysate (composition: 65 g L−1 xylose,
13 g L−1 glucose and 6.3 g L−1 arabinose). Xylitol production started
at 48 h of fermentation and continued up to 144 h. Glucose, how-
ever, was consumed mostly in the first hours of microorganism
incubation, with total assimilation completed by 24 h. Misra et al.
[61] also studied the potential of corncob hydrolysate for xylitol
production using a strain of C. tropicalis. These authors achieved a
maximum extraction of 20.92 g L−1 xylose using 1% (v/v) sulphuric
acid in the corncob hydrolysis. Concentrating the hydrolysate up
to 52.71 g L−1 , in relation to xylose concentration, 15.19 g L−1 of
xylitol was obtained at 60 h of culture. Ping et al. [36] used a concen-
trated non-detoxified corncob hydrolysate for xylitol production by
C. tropicalis CCTCC M2012462, reaching a maximum of 38.8 g L−1
xylitol.
Barley straw was evaluated for its employment in xylitol pro-
duction by a genetically modified T. reesei [49]. Two hydrolysis
methods were tested in the raw material (one by adding organic
solvents and the other by addition of NaOH). The best xylitol pro-
duction (13.22 g L−1 ) was achieved with the use of barley straw
treated with organic solvents plus the addition of 2% xylose.
Detoxified bamboo culm acid hydrolysate (19 g L−1 xylose) was
fermented by C. magnoliae FERM P-16522 in a study by Miura
et al. [9]. The authors [9] concluded that this raw material has the
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potential to be utilized in xylitol production, reaching a maximum
concentration of 10.5 g L−1 for the polyol.
Martinez et al. [11] investigated the production of xylitol
and ethanol by Hansenula polymorpha from sunflower stems
hydrolysed with phosphoric acid. The results of this research
showed that, for an initial concentration of total reducing sugars
of 13.3 g L−1 obtained in the process, xylitol and ethanol yields
were 0.023 g g−1 and 0.14 g g−1 , respectively. Rocha et al. [38],
in a study on ethanol production from non-detoxified cashew
bagasse hydrolysate (29.08 g L−1 glucose, 24.48 g L−1 xylose and
11.33 g L−1 arabinose), observed simultaneous xylitol production
by the yeast K. marxianus CE025. Xylitol production was high-
est at 72 h of fermentation, reaching a concentration of 4.77 g L−1 .
Some recent researches have also been dedicated to evaluating
xylitol production in synthetic media containing xylose as the
main carbon source. Srivani and Setty [66] studied the optimiza-
tion of several parameters (pH, temperature and initial xylose
concentration) for xylitol production from xylose fermentation
by C. parapsilosis NCIM-3323. It was concluded that the maxi-
mum production of xylitol (28.14 g L−1 ) was achieved with the
following values for pH, temperature and initial xylose con-
centration: 3.5, 30 ◦ C and 60 g L−1 , respectively. Vajzovic et al.
[74] used a synthetic medium (containing glucose or xylose,
30 g L−1 ) to evaluate xylitol production in the presence of cer-
tain inhibitors (furfural, 5-hydroxymethylfurfural and acetic acid)
and the results obtained by these authors showed that high con-
centrations of inhibitors (above 3 g L−1 ) negatively affected xylitol
production.
3.3.4. Nitrogen sources
Various nitrogen sources are being investigated in biotechno-
logical studies to optimize the growth of microorganisms and the
production of metabolites of interest. Among the most studied
organic sources are peptone, yeast extract and casamino acid [75].
Among the inorganic sources are ammonium sulphate and phos-
phate, sodium nitrate and urea [9,41].
There have been many studies investigating the influence
of urea utilization as a nitrogen source for xylitol production.
Rodrigues et al. [76] investigated the effects of employing urea
and ammonium sulphate as nitrogen sources on xylitol pro-
duction by Pichia stipitis YS-30. The medium, containing corn
stover hydrolysate, was supplemented with either 5.0 g L−1 urea
or 5.0 g L−1 ammonium sulphate. It was observed that when urea
was used instead of (NH4 )2 SO4 , xylose consumption and ethanol
production rates increased by 25% and 34%, respectively.
Ko et al. [77] studied xylitol production employing waste wood
fermentation. The use of urea as a nitrogen source in the fermenta-
tion medium was tested, replacing the yeast extract with 10 g L−1
urea or 10 g L−1 soybean meal. The yield obtained using urea was
1.3-fold higher than that obtained with yeast extract. Zhang et al.
[10] evaluated ethanol and xylitol production from C. athensensis
SB18 from vegetable waste, supplementing the production medium
with 2.0 g L−1 urea. Significant variations in the parameters of
biomass growth and xylitol production were observed. The strain
preferably used urea as a nitrogen source and showed high conver-
sion of xylose to xylitol, which was completely consumed in 108 h
of fermentation. The highest cell density and xylitol production
were 12.07 g L−1 and 112.27 g L−1 , respectively. The results indi-
cated that, among the nitrogen sources investigated, urea is the
most promising for the studied strain.
In contrast, Hongzhi et al. [78] conducted an experimental sta-
tistical design to optimize a culture medium for xylitol production
from corn bagasse hydrolysate. The nitrogen sources used in the
experiments were ammonium nitrate, peptone, urea, ammonium
sulphate and yeast extract. Among these sources, only (NH4 )2 SO4
Please cite this article in press as: Albuquerque TLd, et al. Biotechnological production of xylitol from lignocellulosic wastes: A review.
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and yeast extract had a significant influence, with optimal concen-
trations of these components of 5.0 g L−1 and 4.6 g L−1 , respectively.
3.3.5. Bioreactor operation modes
Most studies about the xylitol production showed in the litera-
ture are conducted in batch mode (flasks or bioreactors). However,
some experiments approach the production of xylitol in other oper-
ating modes, as in fed batch mode (Table 1), which can achieve
higher yields.
Silva et al. [79], for example, used fluidized bed reactor operated
in semi-continuous mode for xylose-to-xylitol conversion by C.
guilliermondii immobilized on porous glass. The reactor was loaded
with 200 g of immobilized system and operated at a volume of
1.4 L in semi-continuous mode. The fermentation was performed
in seven cycles (total time of 672 h), using fresh medium for each
one. The first and second cycles provided the highest values of
xylitol yield of 0.79 g g−1 and 0.57 g g−1 , respectively, and volumet-
ric productivity of 0.52 g L−1 h−1 and 0.60 g L−1 h−1 , respectively, a
decrease being observed afterwards.
Santos et al. [80] studied the influence of aeration rate and car-
rier concentration on xylitol production from sugarcane bagasse
hydrolysate in immobilized-cell fluidized bed reactor. The results
obtained showed that carrier concentration (Cs) had a negative
influence on xylitol yield based in the consumption of xylose
(YP/S ) and volumetric productivity (QP ), whereas aeration rate (AR)
had a positive influence on QP and a negative influence on YP/S .
With an AR = 0.093 min−1 and Cs = 62.5 g L−1 , the YP/S value was
low (0.25 g g−1 ), but the QP was highest (0.44 g L−1 h−1 ), probably
because cell metabolism was faster when more oxygen was avail-
able to the yeast. When the highest levels of the AR and Cs were
used, the xylitol yield and concentration were the lowest and the
cell concentration was the highest, suggesting that cell metabolism
was directed preferentially towards biomass production instead of
xylitol accumulation.
Salgado et al. [81] evaluated the coupling of two sizes of
CSTR-type bioreactors for sequential lactic acid and xylitol pro-
duction from hemicellulosic hydrolysates of vineshoot trimmings.
By connecting two reactors of 2 L and 10 L, operational condi-
tions were set up for the sequential production of lactic acid and
xylitol in continuous process, considering the dependence of the
main metabolites and process parameters on the dilution rate.
The maximum production of xylitol (5.1 g L−1 ), volumetric xyli-
tol productivity (QP = 0.218 g L−1 h−1 ), volumetric rate of xylose
consumption (QS = 0.398 g L−1 h−1 ) and product yield (0.55 g g−1 )
were achieved at an intermediate dilution rate of 0.043 h−1
(F = 3.55 mL min−1 ).
3.4. Xylitol purification from fermented hemicellulosic
hydrolysates
In recent years, xylitol crystallization has attracted the attention
of scientists around the world. Recovery and purification are com-
plicated steps in many fermentation processes, since they usually
depend on both the nature of the product and the complex com-
position of the fermentation broth. Xylitol, sorbitol and arabinitol
are difficult to separate on an industrial scale and a specific sepa-
ration technique would be of great value. Regarding polyols, very
little information is available on xylitol recovery, with reports more
oriented to the treatment of hemicellulosic hydrolysates, fermen-
tation and metabolic bioconversion [82].
Crystallization is a first-order phase transition. In other words, at
the transition point the solid and liquid phases are in equilibrium;
a supersaturated or undersaturated liquid phase can be created
by a change in concentration, temperature or pressure; the two
phases are separated by a crystal surface with an interfacial tension
higher than zero; and supersaturation is needed to overcome the
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nucleation barrier caused by the interfacial tension [83]. Crystal-
lization is a separation process where a solid phase is created from
a liquid phase. About 70% of the industrial products sold as bulk
chemicals, intermediates, fine chemicals, biochemicals and food
additives are solids [83].
The literature reveals few studies on recovering xylitol from
fermented broths, and the establishment of a methodology that
allows recovery and makes its biotechnological production viable
is desired. Many studies have attempted to purify xylitol using a
crystallization process. The prevalence of crystallization as a purifi-
cation method is due to the feasibility of obtaining high-purity and
uniformly sized crystals, which facilitates the following steps in the
process such as filtration, washing and drying of the product.
In order to improve the purity and yield of xylitol crystals
some studies have focused on the purification process, using ion-
exchange resins and optimization of crystallization conditions.
Xylitol purification from fermented broths is difficult because of
the low product concentration and the complex composition of
the media. Even though crystallization has been employed to
recover xylitol from fermentation media formulated with commer-
cial xylose, the complex composition of cultivated hydrolysates
does not allow a direct crystallization, requiring previous purifi-
cation steps such as adsorption on activated charcoal and ethanol
precipitation [84].
Wei et al. [85], for example, evaluated the crystallization and
purification of xylitol from fermented corncob hydrolysate. The fer-
mented broth was initially bleached with activated carbon (solid
percentage 1%, 60 ◦ C, 165 rpm), desalted with a combination of
two ion-exchange resins (732 and D301x), and residual sugars
were separated using resin UBK-555 (Ca2+ ). Then, the solution
was concentrated in vacuum to supersaturation (750 g L−1 xylitol).
Afterwards, 1% xylitol crystals were added to the supersaturated
solution, which was then cooled to −20 ◦ C for 48 h. Crystalline
xylitol with a purity of 95% and a yield of 60.2% was obtained
by crystallization from the clarified fermentation broth. Mussatto
et al. [86] studied xylitol purification from hemicellulosic sugar-
cane bagasse hydrolysate fermented by C. guilliermondii FTI 20037.
Undesirable impurities were extracted from the broth using ethyl
acetate, chloroform or dichloromethane, with the best results for
clarification of the fermentation broth obtained with ethyl acetate,
also considering xylitol loss. When ethanol, acetone or tetrahydro-
furan were used to precipitate impurities, only tetrahydrofuran was
effective in clarifying the fermentation broth but a large loss in
xylitol content was noted (about 30%).
Rivas et al. [84] evaluated the purification of xylitol obtained by
fermentation of corncob hydrolysates by a strain of Debaryomices
hansenii. Under selected conditions, 43.7% of xylitol contained in
the initial fermentation broth was recovered in well-formed, homo-
geneous crystals, in which xylitol accounted for 98.9% of the total
oven-dry weight.
4. Future prospects and conclusions
This article reviews the production of xylitol and the interest
in xylitol has increased considerably in recent years, due to many
commercial applications in different industrial sectors like food,
dental related products, and pharmaceuticals. Therefore, over the
past few decades much effort has been devoted to the development
of cost-effective and environmentally friendly biotechnological
processes by evaluating lignocellulosic substrates. Numerous
types of hydrolysates of low-value raw materials have been used
for biotechnological production. Additionally, a wide variety of
microorganisms is currently being assessed for their ability to pro-
duce the polyol from these culture media, especially yeasts from
several genera, achieving significant productivity. The scientific
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9
literature presents few researches related to the economic viability
of the biotechnological production of xylitol from lignocellulosic
waste, therefore, there is a need for further research to show
data referents to this topic. The recovery of xylitol from lignocel-
lulosic media fermented by microorganisms is poorly explored
in scientific papers, as well as the possible viable techniques for
downstream processes. Theses processes are one of the most
essential aspects for development of integrated technological
solution for production of second generation biorefinary products
like xylitol via biotechnological process at an economic industrial
scale. Finally, there are limited data in the literature about the
parameters of scaling-up the xylitol production and modes of
reactors operation that can be used to optimize this process.
Acknowledgments
The authors are grateful for the financial support provided by
the Brazilian research agencies CNPq and CAPES.
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