Appl Biochem Biotechnol (2009) 155:356–365

DOI 10.1007/s12010-008-8458-y

Banana as Adjunct in Beer Production: Applicability

and Performance of Fermentative Parameters

Giovani B. M. Carvalho & Daniel P. Silva &

Camila V. Bento & António A. Vicente & José A. Teixeira &
Maria das Graças A. Felipe & João B. Almeida e Silva

Received: 22 May 2008 / Accepted: 26 November 2008 /

Published online: 17 December 2008
# Humana Press 2008

Abstract Traditionally, the raw materials for beer production are barley, hops, water, and
yeast, but most brewers use also different adjuncts. During the alcoholic fermentation, the
contribution of aroma compounds from other ingredients to the final beer flavor depends on
the wort composition, on the yeast strain, and mainly on the process conditions. In this
context, banana can also be a raw material favorable to alcoholic fermentation being rich in
carbohydrates and minerals and providing low acidity. In this work, the objective was to
evaluate the performance of wort adjusted with banana juice in different concentrations. For
this, static fermentations were conducted at 15 °C at pilot scale (140 L of medium). The
addition of banana that changed the concentration of all-malt wort from 10 °P to 12 and
15 °P were evaluated (°P is the weight of the extract or the sugar equivalent in 100 g
solution, at 20 °C). The results showed an increase in ethanol production, with
approximately 0.4 g/g ethanol yield and 0.6 g/L h volumetric productivity after 84 h of
processing when concentrated wort was used. Thus, it was concluded that banana can be
used as an adjunct in brewing methods, helping in the development of new products as well
as in obtaining concentrated worts.

Keywords Beer . Raw materials . Adjunct . Banana . Brewing . Fermentation

Introduction

Traditionally, the raw materials for beer production are malted grains of barley, hops, water,
and yeast, but most brewers add different adjuncts. Conventional beer fermentation

G. B. M. Carvalho : C. V. Bento : M. G. A. Felipe : J. B. Almeida e Silva

Biotechnology Department, Engineering School of Lorena, University of São Paulo, Campus I,
P.O. Box 116, 12602-810 Lorena, SP, Brazil

D. P. Silva (*) : A. A. Vicente : J. A. Teixeira

Centre of Biological Engineering, IBB—Institute for Biotechnology and Bioengineering,
University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal
e-mail: silvadp@deb.uminho.pt
e-mail: silvadp@hotmail.com
Appl Biochem Biotechnol (2009) 155:356–365 357

technology is based on batch fermentation of wort, with no mechanical stirring and with
specific temperature profiles during fermentation to obtain the desired characteristics, such
as ethanol concentration, taste, and aroma on the final product. According to Linko et al.
[1], beer production consists of four stages: (a) malting (based on germination and treatment
of barley); (b) mashing or wort production (extraction and hydrolysis of the components of
malt and possibly other cereals, followed by separation of non-soluble components and
boiling with hops); (c) fermentation stages (in most cases divided into main fermentation
and lagering or maturation); and (d) down-stream processing (filtration, stabilization,
bottling, and other treatments of the final product; Fig. 1).
Presently, there are many variations of this brewing process due to improvements in
technical, biochemical, microbiological, and genetic inventions implemented by the modern
brewing industry [1–5]. Previously, we evaluated a new technology of high gravity brewing
for selectively producing beers from more concentrated wort (using high maltose syrup)
with the goal of producing good quality beer with less time and expense [6–9]. In an
industrial context, traditional brewing uses worts of 10–12 °P to produce beers with 4–5%
v/v ethanol content, while the high gravity technique involves preparation and fermentation
of media higher than 12 °P (degrees Plato, which is the weight of the extract or the sugar
equivalent in 100 g of the solution at 20 °C). By increasing wort concentration, after

Fig. 1 Simplified scheme No-Malted Materials

for brewing, adapted from Linko Barley (Adjuncts)
et al. [1]

Malting Malt Pretreatment

Mashing

Separation

Hops Boiling

Wort

Yeast Main
Fermentation

Maturation

Beer
Down-Stream
Processing
358 Appl Biochem Biotechnol (2009) 155:356–365

preparation, the beer is adjusted to the desired ethanol concentration with oxygen-free water
producing higher ethanol yields more efficiently [10–13].
Recently, new techniques and processes have been developed using several adjuncts for
production of different beers such as alcohol-free, low-alcohol, or aromatic beers [14–16].
Banana fruits are produced in large quantities in tropical and subtropical areas. Besides
being one of the most significant foodstuffs in the world, banana is one of the oldest fruits
known to mankind as an important energy-producing food, while being also a good source
of mineral salts and vitamins [17]. Banana is a general term embracing a number of species
or hybrids in the genus Musa of the family Musaceae, which are native to Southeast Asia
[18]. Today, Cavendish subgroup banana cultivars (M. cavendishii) are the mainstays of the
export trades [18]. However, as is the case for most tropical products, due to the special
climatic conditions needed to grow bananas, they are mainly produced in developing
countries. According to the Food and Agriculture Organization of the United Nations [19],
India produced the most with 11.7×106 tonnes in 2006, which corresponded to 16.5% of
the worldwide production, and Brazil was the second with 7.1×106 tonnes (approximately
10% of the worldwide production).
Banana can also be a raw material for alcoholic fermentation, being rich in
carbohydrates and minerals and providing low acidity. Because of the high yields and
low production costs of bananas, many countries such as Brazil have been utilizing them
widely. In fact, recent research has shown that banana has potential, both from its functional
properties and as an economic activity, for application in processed foods and has become a
commercially viable product in terms of its use as an industrial raw material [17, 20].
According to Viviani and Leal [20], inadequate handling of bananas during the post-
harvest is the main reason for the loss of value of the product and for the loss of export
opportunities. For these reasons, research aimed at applying banana juice as an adjunct in
brewing processes has scientific interest and great potential in many tropical banana-
producing countries.
The goals of this research are to describe and evaluate the effects of the use of banana
juice adjunct and its application in concentrated wort on the performance of fermentation
parameters in a brewing process. Special attention was paid to fermentation performance in
terms of volumetric productivity, ethanol yield, and sugar consumption. The study
evaluated the addition of banana juice to change the concentration of all-malt wort from
10 °P to 12 and 15 °P. There is little or no information on the application or evaluation of
banana adjuncts in high gravity fermentation technology to the brewing of beer in the
literature.

Materials and Methods

Yeast Strain and Inoculum Preparation

The yeast strain used in this work was a commercial lager brewing strain (Saccharomyces
cerevisiae) maintained on malt-agar slants at 4 °C. The yeast biomass for initial inoculation
in the fermentation reactor was cultivated on a rotary shaker (200 mL/30 °C/200 rpm, 18 h),
followed by a 4-L Erlenmeyer flask under static and aerobic conditions (2 L/15 °C/0.1 vvm
of aeration, 30 h), then in a 40-L stainless steel vessel under static and aerobic conditions
(20 L/15 °C/0.01 vvm of aeration, 54–60 h) until the cell concentration was sufficient to
provide a cell concentration of 10–20×106 cells/mL for the fermentation in the pilot tank
(140 L). All inoculum preparations were carried out using all-malt wort (12.5 °P).
Appl Biochem Biotechnol (2009) 155:356–365 359

Wort Production and Fermentation Conditions

Worts were produced according to conventional brewing techniques in the pilot installation used
for the experiments (pilot brewery of the Biotechnology Department of the Engineering School
of Lorena, University of São Paulo USP/Brazil, as shown in Fig. 2). Banana juice, prepared
from fruits of the variety Prata (Musa spp.) and provided by EMATER (Empresa de
Assistência Técnica e Extensão Rural do Estado de Minas Gerais, Cristina-MG/Brazil), was
used to change the concentration of the all-malt wort from 10 °P to 12 and 15 °P. The typical
carbohydrate profile of the banana used in this research (classified by the peel color as ripening
stage 8—yellow with more brown specks, according Loeseck [21]) was 12.4% w/w sugars as
glucose and fructose, 3.2% w/w sugars as sucrose, and 0.5% w/w starch. The banana juice was
produced in a solution of 0.29 g/L by enzymatic treatment (43.2 °C during 1.3 h, with
agitation of 32 rpm, pH 5, and 8.4×10−7 L of Ultra Pectinex SP-L enzymatic pectinolytic
solution by gram of medium) with subsequent thermal treatment (84 °C during 0.67 h) and
mixed to the all-malt wort during the initial stage of wort preparation.
The initial pH value of the obtained wort was adjusted to 5 (by the addition of lactic
acid), and the initial dissolved oxygen concentration was set at approximately 8 mg/L.
Static fermentations were performed in a 180-L cylindrical–conical tank (brewing
fermenter) with 140 L working volume at a constant temperature of 15 °C and wort
concentrations of 12 and 15 °P. A preliminary pilot-scale fermentation trial was conducted
in order to gain an understanding of the conventional brewing process as well as to
determine the performance of fermentations using all-malt wort under the same process
conditions. The fermentation runs were carried out until the apparent attenuation was about
70–75% (1.0 °P above the final value of fermentable sugars).

Analytical Methodology

During fermentations, samples were taken in triplicate and the yeast was removed by
centrifugation at 4,000×g for 20 min. The apparent extract and ethanol concentrations in the
supernatant were measured at 20 °C using an automatic beer analyzer (Beer Analyzer 2,
Anton Paar, Graz, Austria). Part of the supernatant was also filtered through a syringe filter

Fig. 2 Pilot system for wort preparation and beer fermentation (pilot brewery of the Biotechnology
Department at the Engineering School of Lorena, University of São Paulo USP/Brazil): T1 hot water tank, T2
wort treatment tank, T3 filtration tank, T4 boiling tank, P pumps, S spray balls (in-line cleaning system), Ru
refrigeration unit, R1 fermentation/maturation reactor, I and II sampling points or final product, 1 water
supply, 2 malt supply, 3 hops supply, 4 air/O2 supply
360 Appl Biochem Biotechnol (2009) 155:356–365

Fig. 3 Apparent extract

consumption (filled square),
ethanol production (filled
triangle), and yeast in suspension
(empty circle) during all-malt
wort brewing process (main
fermentation) under the following
conditions: initial wort concen-
tration of 12 °P, constant
temperature of 15 °C

(Sep-Pak C18 cartridge, Waters, Milford, MA, USA), and 20 µL was injected into an HPLC
system which consisted of an Aminex HPX-87H column (300×7.8 mm2, Bio-Rad
Laboratories Ltd, Hercules, CA, USA) at 45 °C using a Shimadzu chromatograph LC-
10AD (Shimadzu Co., Tokyo, Japan) with refractive index detector. The mobile phase was
0.005 mol/L H2SO4 at 0.6 mL/min flow rate. Sugar concentrations, reported as glucose,
fructose, maltose, and maltotriose, were determined from calibration curves obtained with
pure compounds.
The yeast cell number was determined using a Neubauer counting chamber, and the
viability was determined by methylene blue staining. All analyses were based on the
techniques described in ASBC [22]. Each determination was made in triplicate.
The specific substrate consumption rates (μSi) were defined as:
mSi ¼ ð1=X Þ
d Si=d t
where Si represents the type of carbohydrate evaluated (maltose, glucose, fructose, or
maltotriose). The derivatives dSi/dt were calculated according to the method proposed by
Le Duy and Zajic [23].
The ethanol productivity (ratio between ethanol produced and total fermentation time, g/L h)
and the ethanol yield coefficient (ratio between produced ethanol and consumed extract, g/g)
were determined after conversion of the apparent extract (°P) to grams of extract per liter of wort
(g/L).

Fig. 4 Percentage of viable cells

(filled triangle) and performance
of S. cerevisiae in suspension
(filled circle) during all-malt wort
brewing process (main fermenta-
tion) under the following condi-
tions: initial wort concentration of
12 °P, constant temperature of
15 °C
Appl Biochem Biotechnol (2009) 155:356–365 361

Fig. 5 Performance of
S. cerevisiae (lager strain) and
percentage of viable cells during
fermentation processes of wort
supplemented with banana juice
under the following conditions:
12 °P/15 °C (cell viability—
empty triangle, and yeast in
suspension—empty circle),
and 15 °P/15 °C (cell viability—
filled triangle and yeast in
suspension—filled circle)

Results and Discussion

In the preliminary pilot-scale fermentation, using all-malt wort, changes in wort gravity,
ethanol production, and cell suspension behavior were evaluated during fermentation at
constant temperature of 15 °C, as shown in Fig. 3. During the first 40 h of fermentation,
substrate consumption (measured in terms of apparent extract) was closely related to the
increase of yeast biomass in suspension. In this interval (0 to 40 h), the apparent extract
decreased by approximately 40%, while the suspended cell concentration increased
fivefold. Concurrently, ethanol production reached a concentration of 2.9% (v/v). However,
at the conclusion of 75.5 h (final fermentation time), the concentration of yeast in
suspension decreased to twice the initial concentration, while the apparent extract
concentration decreased to 3.6 °P and the ethanol concentration increased to 4.7% (v/v).
According to Guido et al. [24], the efficiency of fermentation, the character, and quality of
the final product in the brewing process are intimately linked with the amount and health of
the yeast being pitched. Evaluating and predicting the cell viability as well as the performance
of the brewer’s yeast during the fermentative stage is therefore an important requisite.
Information was obtained in our research by monitoring the percentage of viable cells and the
performance of S. cerevisiae in suspension when using all-malt wort during the main
fermentation (initial wort concentration of 12 °P, at constant temperature of 15 °C; Fig. 4).
The concentration of cells in suspension reached a maximum value of 8.6×107 cell/mL at

Fig. 6 Extract consumption

and ethanol production during
fermentation processes of wort
supplemented with banana juice
under the following conditions:
12 °P/15 °C (apparent extract—
empty square, and ethanol
concentration—empty triangle),
and 15 °P/15 °C (apparent
extract—filled square and ethanol
concentration—filled triangle)
362 Appl Biochem Biotechnol (2009) 155:356–365

Table 1 Control and fermentative parameters used during the main fermentation process of all-malt wort
(12 °P/15 °C) and wort supplemented with banana juice (12 and 15 °P/15 °C): total time (time), final ethanol
concentration (Eth), ethanol volumetric productivity (Pr), and yield coefficient (YP/S), at the end of each
fermentation run.

Fermentation medium (type and concentration) Control and fermentative parameters

Time (h) Eth (% v/v) Pr (g/L h) YP/S (g/g)

All-malt wort process (12 °P) 75.5 4.74 0.50 0.43

Wort with banana adjunct (12 °P) 72.0 5.30 0.58 0.45
Wort with more banana adjunct (15 °P) 84.0 6.39 0.60 0.42

40 h, with high cell viability (100–98%). However, the cell viability decreased to 95–90%,
probably due to the combination of lower substrate concentrations and higher ethanol
concentrations after 40 h. Of the several factors known to affect the yeast fermentative
capacity, the composition of the medium is one of the most important [24].
Following this preliminary fermentation experiment, two trials were conducted in pilot
scale at a constant temperature of 15 °C with concentrations of malt wort from 10 °P to 12
and 15 °P. The cell viability results (in relation to the total number of cells in suspension)
using worts concentrated with banana juice are shown in Fig. 5. There was little variation of
cell viability between 12 and 15 °P concentrations. However, improvement in stability with
higher banana juice concentrations (Fig. 4) indicated a positive effect of the nutrients or
sugar profiles from banana on the viability and performance of brewer’s yeast cells.
Figure 5 also shows that during the first hours of fermentation the number of yeast cells
in suspension increased, then decreased due to flocculation. This profile is in agreement
with that obtained with all-malt wort (Fig. 3). However, in contrast with the trial using all-
malt wort, the fermentation with banana juice adjuncts under the same conditions of
concentration (12 °P) showed a maximum number of yeast in suspension at an earlier stage
of the fermentation and with a more defined profile (Fig. 5). Willaert [25] mentioned that
the biomass peak, resulting from the flocculation of the yeast cells, can influence the total
consumption of the fermentable sugars as maltose and that a higher attenuation with less
raw material loss can be obtained by preventing the sedimentation of the yeast cells.
From the results shown in Fig. 6, it is important to note that the wort with higher initial
extract concentration (15 °P) produced more ethanol than the one with traditional

Fig. 7 Specific substrate

consumption rates (1/h) of the
fermentable sugars during
fermentation process of wort
supplemented with banana juice
(15 °P/15 °C): maltose (empty
square), glucose (empty triangle),
fructose (empty circle), and
maltotriose (empty diamond)
Appl Biochem Biotechnol (2009) 155:356–365 363

Fig. 8 Glucose and fructose

concentrations during fermenta-
tion processes of wort supple-
mented with banana juice
under the following conditions:
12 °P/15 °C (glucose—empty
triangle and fructose—
empty circle), and 15 °P/15 °C
(glucose—filled triangle and
fructose—filled circle)

concentration (12 °P). Table 1 shows the main fermentation parameters obtained for beer
production processes using all-malt wort and wort with banana juice as adjunct. In these
cases, the higher value of fermentable carbohydrate concentration due to the banana juice
adjunct may be the reason for the higher ethanol volumetric productivities. There is,
however, evidence to show that when the banana juice is used as adjunct in the brewing
wort, the efficiency of fermentation may be increased. The different nutrients found in the
banana juice together with the fermentable sugars present in this type of extract may be the
reason for this behavior of the system.
According to Navarro et al. [26], the sugars in wort are not all fermented equally well.
Since the yeast has to hydrolyze sugar polymers before it can use them, it always attacks
hexoses first. In our work, as shown in Fig. 7, comparing the specific rates of consumption
for each fermentable sugar found in concentrated wort with banana juice as adjunct (15 °P),
the following order of assimilation was observed: glucose was consumed first, followed by
fructose, maltose, and finally maltotriose. The uptake and consumption of maltose and
maltotriose was repressed or partially inactivated due to elevated glucose concentration.
According to Willaert [25], only after 60% of the wort glucose has been used by the yeast,
can maltose be consumed. Willaert [25] also showed that maltotriose uptake is inhibited by

Fig. 9 Maltose and maltotriose

concentrations during fermenta-
tion processes of wort supple-
mented with banana juice under
the following conditions: 12 °P/
15 °C (maltose—empty square
and maltotriose—empty
diamond), and 15 °P/15 °C
(maltose—filled square and
maltotriose—filled diamond)
364 Appl Biochem Biotechnol (2009) 155:356–365

high glucose and maltose concentrations, which is in agreement with the behavior found in
this research.
Figure 8 shows that the main consumption of glucose and fructose in the wort fermentation
of 12 and 15 °P with banana juice as adjunct occurred approximately at 38 and 45 h,
respectively. The difference in the consumption profile of these sugars was probably due to
the difference in the initial concentration, which in turn was due to the use of different
concentrations of banana juice in the initial worts. However, as the preparation conditions and
amounts of all-malt wort were kept constant, the consumption behavior of maltose and
maltotriose was nearly almost independent of the wort concentrations used (Fig. 9). The
difference is in agreement to the delay in the consumption of one of the sugars (the fructose)
or of both (glucose and fructose) when using higher wort concentration (15 °P).
The color of the wort produced with banana juice as adjunct is slightly darker than the
one obtained from all-malt worts or from traditional commercial processes. At present, it is
still not clear whether there is a direct relationship between banana concentration as adjunct
and color development during fermentation stage. However, it is very likely that enzymes
present in the fruits (such as polyphenol oxydase) will contribute to the appearance of a
darker color (enzymatic browning).
It is well known that the fermentation of worts of high specific gravity often leads to
unbalanced flavor profiles [27]. Furthermore, in this type of process, the relative amounts of
different assimilable sugars in wort also have an influence on trace compounds extremely
important for the flavor profile in general. Worts containing higher glucose and fructose
produce more esters than worts with high maltose contents [28–30]. The reason for the
difference in ester production between glucose- or maltose-grown cells is still unclear.
Thus, as the ester production is responsible for the fruity character of fermented beverages,
volatile esters constitute an important group of aromatic compounds in beer. In modern high
gravity fermentations, which are performed in tall cylindroconical vessels, the beer ester
balance is often suboptimal, resulting in a clear decrease in beer quality [27]. Thus, when
using banana as adjunct in the brewing process, some advantage in this sense is expected.
The use of banana adjuncts may improve processes that improve beer quality.

Conclusions

For the fermentation conditions evaluated in this research, increases in the initial sugar
concentration of brewing all-malt wort from 10 °P to 12 and 15 °P by using banana juice
adjuncts increased in ethanol production, with volumetric productivity of approximately
0.6 g/(L h) and an ethanol yield coefficient close to conventional conditions (≈0.4 g/g, YP/S).
It can be concluded that banana juice used as adjunct in brewing methods aids in the
development of new and different brewing products (by the use of simple techniques of
preparation of banana juice). However, further work is needed to provide an understanding of
the organoleptic profile obtained during the process of beer production using banana juice as
adjunct.

Acknowledgments The authors acknowledge the financial support from Fapesp (Fundação de Amparo à
Pesquisa do Estado de São Paulo/Brasil), CAPES (Coordenação para Aperfeiçoamento do Ensino Superior/
Brasil), GRICES (Gabinete de Relações Internacionais da Ciência e do Ensino Superior/Portugal), and FCT
(Fundação para a Ciência e Tecnologia/Portugal), as well as the assistance from Malteria do Vale, Wallerstein
Industrial & Commercial, Johnson-Diversey, Novozymes, and EMATER-MG for supplying brewing
materials and banana fruits.
Appl Biochem Biotechnol (2009) 155:356–365 365

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