Fapurusi: Sugar Changes during Preparation of Burukutu Beer 79

density (1 ‘4)and it is readily available within the dried fruit C.S.I.R.O.,

industry. Division of Horticultural Research,
Owing to the simplicity of the gritometer, the cost of Glen Osmond, South Australia 5064
production and maintenance is low and operators need have and
no special skills. Furthermore the few steps involved in the Merbein,
procedure allow determinations to be carried out quickly. Victoria, Australia
After a few days’ experience, operators in Packing Houses can Received 28 July, 1969
average 10 estimations per hour.
Siro gritometers are currently being used in most dried
fruit Packing Houses in Austrialia. References
1. Cruess, W. V., ‘Commercial fruit and vegetable products’,
Acknowledgments p. 593 (New York: McGraw-Hill)
2. Moyer, J. C., ‘Dirt content of grapes’, Research Circular No. 6,
The authors thank Miss S. Oliver and Miss J. Evans for 1966, (Geneva, N.Y.: New York State Agric. Exp. Stn)
valuable technical assistance, Mr. P. Johnston for construction 3. Hawker, J. S., & Grncarevic, M., Fd Technol. A m . , 1968, 20,
work, the Australian Dried Vine Fruits Research Committee 218
4. Aust. P., Appln No. 39619168
for financial support and Mr. K. M. Cellier of the C.S.I.R.O., 5. Harris, J., Grncarevic, M., & Hawker, J. S., Fd Techno[. Ausr.,
Division of Mathematical Statistics for statistical analyses. 1968,20, 537

SUGAR CHANGES DURING THE PREPARATION

OF BURUKUTU BEER
By S. I. FAPARUSI*

In the study of sugar changes during preparation of Burukutu beer, samples analysed were obtained from
a local brewer. A freshly prepared drink was analysed together with gari - a starch adjunct. The
pattern of sugar changes during this 5-day malting stage was also followed. Twelve sugars were identified
in the beverage while six sugars were shown to be present in the gari adjunct. Analyses of both germinated
and ungerminated sorghum grains show that no new types of sugars resulted from the process of germina-
tion. In the study of amylase activity of the malts it was shown that detectable amylase activity began
from the second day of germination onwards. Because of the relatively high concentrations of malto-
oligosaccharides in the drink, starch amylosis should have continued during the fermentation stage.

Introduction The mixture is left to ferment for 2 days. At the end of

Burukutu is an alcoholic beverage brewed from the guinea
corn grains (Sorghum vulgare; = S. bicolor (Moench)); in the
savannah regions of Nigeria. During the preparation,
sorghum grains are steeped in water overnight. The grains
are then poured into a basket and the water is allowed to
drain off for about 2 hours.
The grains are spread out on leaves or mats in a bed of
about 3-4 in. in thickness and covered with another layer of
leaves. During this malting period, the grains are watered
on alternate days and occasionally turned over.
Germination, which starts within 24 h after steeping, is
allowed to continue for a period of 4-5 days. The length of
plumule is used in judging when germination has gone far
enough. However, germination is usually complete after a
period of 4 days.
After germination the malt is spread in thin layers in the
sun to dry for 1 or 2 days. The dried malt is ground into
powder.
In the next stage, gari (a starchy powder produced from
the tuber of cassava plant, Manihot utilissima Pohl.) is added
to a mixture of the ground malt and water and stirred with a
stick. The resulting mixture (gari-malt powder-water) is
roughly in the ratio 1 : 2 : 6 by vol.
* Present address: Department of Brewing and Biochemistry,
Heriot-Watt University, Edinburgh
J. Sci. Fd Agric., 1970, Vol. 21, February
fermentation, the mixture is boiled for about 4 h. The drink
is then left to mature for another period of 2 days. The end
product is a suspension of some particles in a creamy liquid.
Similar alcoholic beverages produced from cereal grains
have been described in other parts of the world, e.g. in the
Republic of South Africa, a kaffir beer is produced from
Sorghum cafrorum Beauv.lT2 In Nigeria little study has been
made on any of the beverages produced from the cereals.
Recently, a study of the microflora of Pito drink was started
(Ekundayo, J. A., personal communication). Pito is a
beverage which can be brewed from millet or corn or sorghum
grains.
The present study was undertaken to determine the different
types of sugars in the Burukutu beer. The changes which
occur in the various sugar concentrations were followed
during the malting stage in order to ascertain the type and
amount of sugars formed during the germination of the
grains.
Experimental
Preparation of sorghum malt
All the samples analysed were obtained from a local
brewer, who bought the sorghum grains from the local
markets. The grains were steeped in water overnight (about
16 h). The water was allowed to drain off from a basket for
about 1 h.
80 Fapurusi: Sugar Changes during Preparation of Burukutu Beer

The grains were then spread out in a bed of up to 3 in. in
thickness and covered with banana leaves and sacks. Each
day samples were taken out from the germinating grains and
spread out in thin layers to dry in the sun.
The remaining grains were watered and turned over every
day. The dried malt was ground in a Waring blender.
Germination was allowed to continue for five days.
Extraction of sugars from the samples
5.0 g of ground malt were weighed into a 100 ml conical
flask. The sugars were extracted with three 50 ml portions
of 80% ethanol on a boiling water bath. Each portion was
extracted for 1 h. Three such extractions were shown from
the preliminary experiments, to be sufficient for complete
extraction of the sugars.
The solid particles in the extract were centrifuged and the
supernatant concentrated in vacuo at a temperature of 60" to
a volume of about 20 ml. The proteins were precipitated
with a suspension of basic lead acetate in water. The red-
brown pigment got from the coat of the grains was also ad-
sorbed by the thick precipitate formed. After centrifuging
the precipitate the lead ions were removed by two successive
bubblings of H2S gas through the solution.
H2S was boiled off and the volume of the extract was
reduced in vacuo at a temperature of 60" to about 3 . 0 ml.
The total volume was made up to 5 .O ml with 20 % (by vol.)
aqueous isopropanol.
Sugar analysis
0.01 ml of each sugar extract was spotted in triplicate on
Whatman No. 1 chromatography paper. The chromato-
gram was developed for 72 h in 1-butanol-acetic acid-water
(4 : 1 : 1 by vol.) using a descending paper chromatographic
method.
TABLEI
Analysis of sugars in gari
Sugar Conc., %by wt.
Mannose 0.90
Galactose 0.50
Maltose 0.32
Maltotriose 0.30
Maltotetraose 0-09
Raflinose 0.45
After drying, one of the triplicate columns was cut off and
the sugars on it were located using the method described by
Dube & Nordin.3 Then, areas containing the spots were
marked out, and corresponding areas on each of the remaining
duplicates were removed and eluted for 3 h with 5 * O ml dis-
tilled water.
To 2.0 ml of an aqueous eluate was added 0.05 ml of 80%
(wt./vol.) aqueous phenol reagent followed by a rapid
addition of 5.0 ml of conc. HzS04. After the sample had
stood at room temperature (25") for 30 min, the optical
density was determined at 480 nm. The concentration of
each sugar was calculated from the standard curve. All
samples were read against a blank containing distilled water
in place of the sugar solution.'L
Assay of amylases
100 ml of 0.2% (wt./vol.) calcium acetate at pH 4.6 were
used to extract the amylases from 10 g of the ground sorghum
malt for 3 hours.s The extract was centrifuged at 12,000
rev/min for 30 min at a temperature of between - 1O and 1". +
The clear supernatant extract was assayed at 37" using a
modified method of Street & Closes as described by Street.6
After the preliminary experiments it was considered essential
to dilute the extract 10 times. The extract was always diluted
with the calcium acetate solution.
Results and Discussion
Table I shows the sugar content of the gari adjunct. The
sugar content in the Burukutu beer is shown in Table 11.
The sugars identified in the beverage were fructose, mannose,
galactose, glucose, galacturonic acid, sucrose, maltose,
isomaltose, raffinose, maltotriose, stachyose and malto-
tetraose.
Fig. 1 shows the variation of sugars with the number of
days of the germination of the sorghum grains. The pattern
of variation of the sugars during this malting stage showed
that the sugars could be divided into three distinct groups:
those sugars which decreased in concentrations on the first
day of germination and then showed a rapid increase in con-
centration on the second day until the third day after which
there was no further substantial increase in concentration,
e.g. glucose and sucrose; those sugars which showed a steady
increase in their concentrations until the second day, after

TABLEI1
Analysis of sugars in Burukutu beer
*-O t
The samples were analysed immediately
after the 2-day maturing period

Sugar Conc., %by wt.

Fructose 0.20
Mannose 0-65
Glucose 0.60
Galactose 0.36
Galacturonic acid 0.48
Sucrose 0.20
Maltose 0.42 0 I 1 I
Isomaltose 0.38 1 3 5
Raffinose 0.60 DAYS OF GERMINATION
Maltotriose 0.56
Stachyose 0.43 FIG.1. Concentration of sugar vs. days of germination
Maltotetraose 0.16 A Sucrose; 0 glucose; 0 fructose; 0 raffinose; A maltose; x maltotriosc;
W isomaltose; V stachyose
J. Sci. Fd Agric., 1970, Vol. 21, February
 Faparusi: Sugar Changes during Preparation of Burukutu Beer
which there was a relatively sharp rise in the concentrations
but they did not increase after the third day, e.g. fructose,
maltose, isomaltose and maltotriose; and those sugars which
did not show any increase in their concentrations, e.g.
raffinose and stachyose.
Previous analyses of sorghum grains have shown the
presence of a number of sugars. Von Holdt & Brand7
reported that only fructose, glucose and sucrose were present
in ungerminated sorghum grains. They also showed, in
addition to these three sugars, that maltose, isomaltose,
maltotriose and traces of higher malto-oligosaccharides were
present in germinated grains. Nordins reported the presence
of stachyose in sorghum grains.
The purpose of the malting stage is to effect the hydrolysis
of starch to fermentable sugars. It is the view of many
workers that amylases are responsible for this process of
hydrolysis. There have been conflicting views as to when the
activity of these enzymes becomes manifested. Novellie2
concluded that the enzymes are synthesised during the
germination stage. However, Dube & Nordin3 reported that
the amylases are usually present in the insoluble condition in
the grains even before germination begins. Here, there is
evidence (Fig. 2) that detectable amylase activity started from
the second day.
During the steeping stage the metabolic activities of the
grains which were previously dormant, were resumed. Some
of the sugars would be used in the metabolic processes.
Sorghum grains germinate rapidly and hence will require high
energy-level compounds. This could be responsible for the
initial fall in the concentrations of some of the sugar. How-
ever, when the amylase activity resumes, more sugars will be
produced than is required for metabolism; thus the concen-
trations of these sugars increase.
It appears that all these sugars were already present in the
grains before germination started. Some suggestions have
been made that the moulds which sometimes grow on the
grains, produce amylases which would attack the ~ t a r c h . ~
This initial sugar production cannot be excluded because
Nigeria is a humid country and moulds grow readily on food
substances.
The local brewers add gari to the mash to increase the
viscosity of the drink. A consumer regards a thin drink as
being too dilute and is therefore considered to be of an
inferior quality.
Analysis of the gari powder (Table I) showed that mannose,
galactose, maltose, raffinose, maltotriose and maltotetraose
were present. Thus, only galacturonic acid was not identified
free in any of the ingredients used in the preparation of the
drink. Galacturonic acid is usually obtained by enzymic
hydrolysis of pectin in plant materials. It is expected that
pectic substance in gari could have been hydrolysed by the
enzymes of the microflora which ferment the drink.
From Table I1 it can be seen that there are relatively high
concentrations of the malto-oligosaccharides, in the drink.
Since the mash was not boiled the amylases of the malt
remained active and a certain degree of amylosis could occur
during fermentation. Also the micro-organisms responsible
for this fermentation could produce amylases which could
hydrolyse the starch. Novellielo reported such a starch
amylosis during kaffir beer fermentation.
During the preparation of gari the starch should have
undergone some degree of gelatinisation. This would make
the breakdown of the starch rapid and thus more sugars
J. Sci. Fd Agric., 1970, Vol. 21, February
81
FIG.2. Graph of amyalse activity against days of germination
Enzyme activity is expressed in Street-Close units
would be produced.ll Because some of the sugars which
arise from starch hydrolysis, e.g. glucose and maltose, could
be used up in the fermentation process while the malto-
oligosaccharides are unfermented, it is expected that these
maltosugars would remain in relatively high concentrations.
Sucrose appears to be the sugar used mainly in the fermenta-
tion. This is not surprising since some of the micro-
organisms which could be responsible for the fermentation of
the drink usually ferment sucrose in preference to other
sugars. Although these micro-organisms have not been
analysed, however, it has recently been shown that Saccharo-
myces cerevisiae is one of the yeasts which ferment Pito drink
(Ekundayo, J. A., personal communication). It is expected
that this yeast would play a major role in this beverage.
Also, from the sour taste of the drink, some lactobacilli may
have played a substantial r d e in the fermentation. This
question will be resolved when the study of the microflora
which ferment Burukutu beer is completed.
Acknowledgment
The author wishes to thank Mme Adamo Yinusa for her co-
operation in preparing the samples analysed in this study.
Biochemistry Department,
University of Ibadan,
Nigeria
Received 24 July. 1969
References
1. Schwartz, H. M., J. Sci. Fd Agric., 1956, 7, 101
2. Novellie, L., J. Sci. Fd Agric., 1960, 11, 408
3. Dube, S. K., & Nordin, P., Archs Biochem. Biophys., 1961,94,
,*,
1Ll
4. Dubois, M., Giles, K. A., Hamiltom, J. K., Rebers, P. A., &
Smith, F., Analyt. Chem., 1956, 28, 350
5. Street, H. V., & Close, J. R., CIinica chim. Acta, 1956, 1, 256
6. Street, H. V., ‘Methods of Enzymatic Analysis’, (Ed. Berg-
mayer, H. U.) 1963 (New York: Academic Press)
I . Von Hodlt, M. W., & Brand, J. C., J. Sci. Fd Agric., 1960, 11,
467
8 . Nordin, P., Chem. Absfr., 1959,55, 8548h
9. Platt, B. S., &Webb, R. A., Proc. Nutr. Soc., 1946,4, 132
10. Novellie, L., J . Sci. Fd Agric., 1966, 17, 354
11. Novellie, L., & Schutte, R. J., J. Sci. Fd Agric., 1961, 12, 552