Biotechnology Letters Vol 7 No I0 743-748 (1985)

SOLID STATE FERMENTATION

OF BANANA WASTES

J. BALDENSPERGER, J. LE MER, L. HANNIBAL and P.J. QUINTO

ORSTOM, Laboratoire de Biotechnologie

et de Microbiologie Appliqu4e
B.P. 81, 97201 FORT DE FRANCE
Martinique (F.W.I.)

SUMMARY

Using a strain of Aspergill~8 niger, the protein content of

banana wastes was raised from 6 to 18 % by solid state fermentation.
A meal of green bananas was obtained by solar drying, and fermentation
was conducted in a 15 kg (dry weight) capacity stirred reactor. As the
substrate consumption was 24 % of initial weight after 43 h. of fer-
mentation, protein production was calculated to be 150 % of the initial
content. According to its composition (50 % total sugars, 13 % reducing
sugars, 18 % proteins) the fermented banana waste could be used as
cattle feed.

INTRODUCTION

In an attempt to deve[op new protein sources in countries

that cannot at present import food or feed because of currency
shortages, a new procedure for solid-state fermentation (Rai~bault and
Germon, French patent 76.06.677) has been studied. It is mainly based
on the use of higher fungi which easily develop in wet but non-liquid
media. According to previous experiments using cassava meal, banana
or potato wastes (Senez et al. 1980), the limiting factor in solid
state fermentation is heat removal. From laboratory results a new
equipment was designed (Deschamps and Meyer, Frency patent 79.02.625)
for a periodically agitated fermentation. The aim of this study was to
evaluate technical feasibility of the agitated pilot fermentor using
banana wastes, a starchy substrate available in many developing
countries (Chung and Meyers, 1979). Due to export regulations, ~5 to
30 % of banana produced are discarded, some of which being destroyed.
In Martinique it represents about 15 OOO tonnes each year, and pro-
tein enrichment of banana wastes could be of economic interest for
cattle feeding. To investigate this possibility experiments were u n d e r -
taken in this tropical country at the pilot scale on solar drying of
banana wastes and the solid state fermentation of the resulting
substrate.

MATERIALS AND METHODS

Aspergillus strain number 2 was previously isolated from cassava,

and evaluated for solid cultivation (Raimbault, 1980). The strain was
maintained on potato-dextrose-agar. Spores were produced in Roux
bottles on the banana meal using the salt solution described by
Raimbault and Alazard (1980) and harvested in 0.2 % tween solution.

743
 l.ruits discarded at the IRFA plantation were sliced (3 mm)
with their peel 2 to 8 days after collection to avoid excessive latex
exudation and ripening. The slices were spread on trays of the experi-
mental solar dryer. This equipment had been developed by our laboratory
for a 20 kg capacity. In fair weather 2 days were necessary to raise
the dry content of banana wastes from 20 to 90 %. The chips were
ground to a fine-grained meal that could be stored several weeks. The
dry matter average composition of the banana meal was 65 % starch, 10 %
reducing sugars, 7 % fibers, 6 % proteins, 5 % minerals. In samples
collected throughout fermentation, total sugars were determined using
anthrone reagent (Viles and S~ilverman, 1949), reducing sugars with
dinitrosalicylic acid reagent (Miller, 1959) and the protein fraction
precipitated by trichloracetic acid was determined with the Folin
phenol reagent (Lowry et al., 1951).

The scheme of the fermentor is presented in Fig. I. The

commercial bread-making blender was modified for steaming and aeration
throughout the perforated bottom. Steam for gelatinization was produced
by an electric boiler. The blender equiped with motors for rotation of
its arm and vessel was fixed upon a weighing machine to control the
weight of the product during fermentation. Temperature was maintained
around optimum (38~ by stirring and spraying with water, the central
control unit regulating the pH and air flow during each stirring cycle.
To check growth rate regulation time was monitored with temperature and
air flow. The pH probe fixed upon a pneumatic jack was pulled out from
the product at the beginning of a stirring cycle, the pH value stored
in the control unit then compared to the set-point, and the pH
correcting solution added if necessary though a spray while stirring
the product. Aeration was provided through a temperature control device
by motored valve operated only during stirring time so that air flow
gradually increased with fermentation time. At the end of each stirrin~
cycle high pressure (9 bars) was connected to the air flow line in order
to clear holes in the bottom of the fermentor vessel.

RESULTS AND DISCUSSION

The data presented were obtained using 8 kgs of banana meal

(90 % dry matter), weighed directly in the vessel. Humidity was raised
to 30 % with a salt solution containing 1OO ml H3P04, 264 g (NH4) 2 S04,
280 g urea in 2.5 liters tap water, this solution being added through a
spray while stirring the meal. Vapor (I bar) was then passed throughout
the substrate during 30 mn, thus rising the temperature up to 80~ The
meal was allowed to cool before addition of the spore solution (2.107
spores/g dry weight) according to Raimbault and Alazard (1980) to a
final humidity of 42 %. This value is significantly lower than that
defined in previous experiments conducted in columns where sticking
of the product does not occur. Trials run at 50 % initial humidity
always failed for this reason. Aeration rate was fixed at 0.8 m3/h with
air heated to 60~ by means of a regulated tank (number 27 in
Fig. I), thus ~iving an incubation temperature of '30"C in the
fermentor. The motored valve (number 22) was set for complete opening
after 4.5 h of stirring time, allowing a maximum air flow rate of 2.4
m3/h. As stirring cycles are controled by tlle product temperature which
results froE~ the balance between heat production by the mold and heat
removal by aeration, opening time of the motored wilve is an important
f e r m e n t a t ion p a r a m e t e r .

744
 |
|

|
"._ ] @
@

Fig. I. Scheme of the stirring solid-state fermentor

I. Modified bread-making blender A2 Jack alimentation

2. Stirring arm motor 19. pH probe jack
3. Vessel rotation motor B. High pressure channel
4. Weighing machine BI Clearing channel
5. Central regulation unit 20. Solenoid valve
6. Recorders (temperature-aeration- 21. Expansion tank
cycles) B2 High-pressure alimentation
7. Temperature probe channel
8. Air compressor 22. Motored valve
9. Drain cock 23. Pressure reducer
10. Pressure reducer B3 Low-pressure-alimentation
A. Low pressure channel channel
AI Spray-alimentations 24. F1owmeter
11. Solenoid valve 25. Solenoid valve
12. Water tank 26. No-return valve
13. Pump 27. Air-heater
14. Water spray C. Vapor channel
15. p}t c o r r e c t i n g solution 28. Steam-boiler
16. pit c o n t r o l unit
17. Urea spray
18. plt p r o b u

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 The initial pH of gelatinized meal co~leaining minerals and
spores was 4.0, which is the set-point fixed for the pH regulation
unit. As mentioned before the urea solution was only sprayed during
part of the stirring time, while the pH probe is out of the solid
medium. Therefore efficiency of pH control depends on proper adjustment
of the pump flow rate, 550 ml/h. in this experiment (P 24).

From previous physiological studies in small devices (IO g

dry weight) it was shown previously that moisture content and incuba-
tion temperature influence germination time (Raimbault and Alazard,
|980). When moisture content was fixed to 50 % or more in the pilot
fermentor, lag phase before the beginning of stirring cycles was
between 12 to 14 h., but we had to reduce drastically the water content
due to the stickiness of the product. At 42 % humidity the lag time was raised
to 24 h. In order to prevent drying out of the surface layer and crust
formation at the bottom, the central control unit started every 2 hours
stirring cycles during the lag phase.

A short acceleration phase (3 h.) during which stirring

cycles occured at shorter intervals followed the germination phase
(24 h.). Then 18 stirring cycles were recorded every hour during 34 h
after w~ich the rate of cycles began to decrease . Fermentation was
stopped 7.5 h later when 9 stirring cycles were recorded every hour. The
fermentor was therefore in action through 68.5 h, corresponding to
24 h and 44.5 h for lag and growth phase respectively . The product was
sampled during the process for analysis and fermentation parameters
were recorded. A set of data for experiment number P24 is reported in
'Fable 1.

Table I. Composition of the fermenting banana meal

Time (hours) O 20 26 28 44 46 48 50 54.5 68.5

Moisture content (%) 42 42 42 43 53 54 55 56 59 62
pH 4.3 4.3 4.3 4.5 3.6 3.6 3.6 3.6 3.8 4.0
Total sugars in 84 84 80 87 70 74 67 47 57 63
sample (% d.w.)
Reducing sugars in 6 13 15 17 26 25 22 21 18 12
sample (% d.w.)
Proteins in sample 6 7.5 8 8 14.5 16 16 15 18 18
(% d.w.)
Total dry weight(kg)7.8 7.7 7.4 7.3 6.b 6.4 6.5 6.4 6.0 5.7
Total proteins (kg) 0.43 0.58 0.58 0.58 0.96 1.O4 1.O3 0.94 1.O7 I.O3

The reactions involved in the solid state fermentation process

of banana wastes are illustrated in Fig. 2, which shows protein produc-
tion, consumption of total and reducing sugars, and total stirring time
during the fermentation. From the figure it [s apparent that fermen-
tation c o u l d have been s t o p p e d a f t e r a t o t a l o f 60 h. (36 h. a f t e r

746
the end of the germination phase). At this time the protein concentra-
tion calculated as a percentage of the initial amount of substrate
reached a maximum of 14 % although the rate o~ total sugar consumption
was 0.4 %/h. Therefore maximum yield of protein production was observed
30 h. after the end of the germination phase.

Concentration of reducing sugars in the fermenting meal

reached its maximum of 25.5 % after 20 h. of growth (44 h. after ino-
culation). With 56 % moisture content the product was not sticky and
kept its granulated structure. However agglomeration of particles in
pellets occured in some previous trials conducted with around 50 %
initial moisture content. Reducing sugar concentrations of 29 %, 28 %,
30 % and even 43 % were respectively measured in such samples after
10 to 15 h. of growth, resulting in complete coagulation of the product
within the vessel. For solid state fermentation of banana meal, a substrate
rich in starch, the accumulation of reducing sugars is a major difficulty
that could be overcome both by limiting available water and by using an
Aspergillus niger strain producing less amylases than Aspergillus hennebergii
which had been chosen for all experiments on potato wastes (Deschamps et
al., 1982).

The regulation time curve is of special interest since it shows

that for a total fermentation time of 68.5 h., the motors for stirring had
to operate only 14 h., i.e. I/5 of the time. Cooling efficiency of stirring
and spraying the product is therefore demonstrated in the pilot fermentor,
and should be considered for its scaling-up.

lIT i5

T Fig. 2. Total and

reducing sugars
consumptions,
protein production
and stirring time
during solid state
fermentation of
banana wastes.

A : total sugars
O : reducing sugars
I : proteins
X : stirring time

FER2ENTATION TIME ( HOURS)

747
 In order to perform nutritional experiments, an emulsion of
vegetable oil in saline solution (360 ml soy bean oils, 60 g calcium
chloride, 60 g monocalcium phosphate, 500 ml tap water) was added
throughout the spray while stirring the fermented product. Therefore
the enriched banana meal could fit with growth requirements for the fresh
water shrimp Macrobrachium rosenbergii which could be a valuable valo-
rization of banana wastes in several countries. The average final compo-
sition of the fermented meal was as follows : 50 % total sugars, |4 %
reducing sugars, 19 % proteins, 12 % ashes, 7 % lipids in the dry
matter. With potassium, calcium and phosphorus contents of respectively
3 %, 3 % and 1.5 % in the dry matter, the balance of minerals was
therefore convenient.

Some preservation experiments have been made, as the fresh

fermented product ( 6 1 % of moisture) could not be stored at room tempe-
rature. In an attempt to kill the fungi, steam was passed throughout,
resulting in blackening and coagulation : steaming does not seem sui-
table as a preservative treatment. The fermented product of one experiment
was ensiled in plastic bags on march 15th and analyzed on june 13th,
when composition of the dry matter was found to be 23 % total sugars,
16 % reducing sugars and 16.5 % proteins. During 3 months 43 % and 32 %
of total sugars and proteins were respectively lost by storage of the
silage at room temperature (25~ the amount of reducing sugars increa-
sing by 7.5 %. Although pH (3.9) was convenient for silage, inoculation
with lactic bacteria should be of interest if this preservative t r e a t -
ment is tO be chosen. Batches used for nutritional experiments with
Macrobrachium rosenbergii were freeze-dryed although this method cannot
be considered for cattle feed production.

As pointed out previously (Hesseltine, |977 ; Moo-Young et al.,

1983) solid state fermentation of agricultural residues could be of in-
terest for producing animal feeds. The pilot scale process described
hereby was successfully applied to green banana wastes. Nevertheless
further work is needed to simplify and scale up the process. Despite
the present economical context in favor of proteins from plant origin,
research on conditioning and utilization of fermented banana meal will
be undertaken considering the availability of this starchy substrate in
developing countries.

REFERENCES
Chung, S.L., and Meyers, S.P. (1979). Bioprotein from Banana Wastes. In:
Developments in Industrial Microbiology.
Deschamps, F., Prebois, J.P., Raimbault, M., Aufeuvre, M., Ansart, M.,
Manguin, H., and Compain, F. (1982). Final Report Convention Ministate
de lrIndustrie. IRCHA, 79.2.34.O510, France.
Hesseltine, C.W. (1977). Process Biochem., 12, 24-27 and 29-32.
Lowry, O.H., Rosebrough, N.J., Farr, A.L,, and Randall, R.J. (1951).
J. Biol. Chem., 193, 265-275.
Miller G.L., (1959). Anal. Biochem., 31, 426-428.
Moo-Young M., Moreira, A.R., and Tengerty, R.P. (|983). Principles of
solid-substrate fermentation. In:The fi~amento~s Fungi, IV,London,Arnold.
Raimbault, M. (1980). Th~se Univ. Toulouse, France.
Raimbault, M., and Alazard, D. (1980). European J. Appl. Microbiol.
BiotechnoZ., 9, 199-209.
Senez,d.C., Raimbault, M., and Deschamps, F. (1980). Rev. Mond. Zootech.
(~O!, 35, 36-39.
Viles, F.J., and Silverman, L. (1949).Ana~. Chem., 21, 950-953.

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