2005/06/04 | Dawadawa 新的一种发酵食品 国内很少人知道啊``
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Flavouring composition prepared by fermentation with Bacillus spp.
M. Beaumont
Nestle Product Technology Centre, New Milford, Connecticut, USA
Accepted 28 December 2000
Abstract
Fermented foodstuffs and condiments remain a key constituents of diets throughout many parts of Asia and Africa. In cases
where the process of fermentation evolved for the development of taste or aroma, it often resulted in enhanced nutrition,
stabilisation of the original raw materials, and detoxification of anti-nutrient factors. Several fermented products rely on the
participation of various Bacillus species, including Bacillus natto and B. subtilis. Often, the finished products are of a very local
character and exhibit sensory properties resulting from unique flora and processing technologies applied in small scale, homebased
fermentations. Fermentation with B. natto and B. subtilis can produce very characteristic aromas in fermented products
such as natto and dawadawa (also referred to as daddawa). Moreover, the hydrolytic capabilities of these microorganisms can
result in a precursor-rich environment, which is useful for subsequent reactions leading to flavour production. A 1995 patented
process demonstrated the ability to produce a fermented flavouring composition with the use of Bacillus spp. Hydrolysed
protein obtained after fermentation with Bacillus spp. is mixed with reactive flavour precursors, which are subsequently heated
to induce flavour formation and can be dried to a powder format. The product of this patented process imparts a basic meaty
flavour, with a reduced yet characteristic dawadawa-like aroma. This paper briefly summarises some of the characteristics and
uses of traditional dawadawa and illustrates alternatives described in the patent for the production of a process flavour base.
Issues and considerations for the industrialisation of a fermentation process are briefly discussed, as well as some future
opportunities for development and exploitation of traditional fermentation technology. D 2002 Elsevier Science B.V. All rights
reserved.
Keywords: Fermentation; Dawadawa; Traditional technologies; Patent process
1. Introduction
1.1. Dawadawa as a traditional culinary product
The fermentation of African locust beans (Parkia
biglobosa) by Bacillus spp. to produce dawadawa is
an example of an alkaline fermentation process
(Steinkraus, 1995). Dawadawa is a culinary product
that can be used to enhance or intensify meatiness in
soups, sauces and other prepared dishes. It is considered
the most important food condiment in the entire
West/Central African Savanna region (Odunfa, 1986).
Other products of alkaline fermentation include Japanese
natto, Thai thua-nao and kinema from India
(Tamang, 1998). The preservative and flavour characteristics
of these fermentations are derived in part
from the liberation of ammonia and increased pH,
0168-1605/02/$ - see front matter D 2002 Elsevier Science B.V. All rights reserved.
PII: S0168-1605(01)00706-1
E-mail address: Mark.Beaumont@rdct.nestle.com
(M. Beaumont).
www.elsevier.com/locate/ijfoodmicro
International Journal of Food Microbiology 75 (2002) 189– 196
concurrent with protein hydrolysis to free amino acids
and peptides. Other African fermentations in which
Bacillus spp. play a role include the production of
Nigerian ugba (fermented African oil bean), Nigerian
ogiri (fermented watermelon seeds) and ogiri-saro
(sesame seed, pumpkin or castor oil seed fermentation)
from Sierra Leone.
It is not the intent of this paper to review in detail
the fermented product or process for the production of
dawadawa, but only to provide a background for
discussion of the patent. It is fortunate that work
exists in the literature on dawadawa microbiology
(Odunfa, 1981; Antai and Ibrahim, 1986; Odunfa and
Oyewole, 1986; Ogbadu and Okagbue, 1988; Odunfa
and Komolafe, 1989; Aderibigbe and Odunfa, 1990;
Allagheny et al., 1996; Olasupo et al., 1996), the
conditions of its processing (Odunfa, 1983, 1985;
Odunfa and Adewuyl, 1985a,b; Ikenebomeh et al.,
1986; Ikenebomeh, 1989; Oyewole and Odunfa,
1990), its nutritional properties (Odunfa, 1983,
1986), as well as alternative applications of the
dawadawa process and raw materials for dawadawa
production (Sarkar et al., 1993; Olasupo et al., 1997;
Amadi et al., 1999). Moreover, some excellent reviews
of dawadawa and related fermentations are
available, particularly from Odunfa (1986), Steinkraus
(1995) and Odunfa and Oyewole (1998). Although
there is no comprehensive report on flavour or dawadawa,
some authors provide insight on the components
contributing to its unique character. (Kanno and
Takamatsu, 1987; Ogbadu et al., 1990; Owens et al.,
1997).
Dawadawa is the name designated by the Hausa
Nigerian tribe, for fermented locust beans (Odunfa,
1986). Similar fermentations have been characterised
throughout Africa, with local adaptation in the form of
raw material selection or (post) processing. These
regional versions are often given local names such
as kinda in Sierra Leone, iru in coastal Nigeria,
soumbara in The Gambia and Burkina Faso and
kpalugu in parts of Ghana (Odunfa, 1986).
1.2. Uses of dawadawa
Dawadawa is primarily used as a condiment or
flavour intensifier for soups and stews and to impart
or enhance meatiness. Although no formal sensory
descriptive analysis of dawadawa was found in a
literature search, those familiar with the product
instantly recognise an unmistakable unique aroma.
This aroma—sometimes described as ammoniacal or
pungent—appears to mellow when dawadawa is used
in cooking applications. A simple culinary application
of dawadawa in the Nestle Product Technology
Centre Kemptthal kitchen (used in an African recipe,
chicken yassa) clearly demonstrated taste modifying
properties of dawadawa which were not easily predicted
from tasting dawadawa on its own. An informal
panel screening of the dawadawa enhanced dish
against a control dish (no dawadawa used in preparation)
indicated that characteristics such as ‘‘cheese’’,
‘‘smoke’’, ‘‘meaty’’ and ‘‘fatty/rancid’’ were associated
with the taste of dawadawa.
1.3. Flavour properties
A recent review of published literature did not
identify any comprehensive analysis on the flavour
of dawadawa. However, the flavour properties of
dawadawa are most likely due to its amino acid
content, in particular glutamate, which contributes to
flavour enhancement, as well as peptides and aroma
volatile constituents. Volatiles may of course be
directly produced during fermentation, or may evolve
as a result of the effect of heat on amino acid and fatty
acid constituents of dawadawa. A study on the characterisation
of aroma volatiles in Bacillus-fermented
soybeans indicated the formation of aroma active
aldehydes, ketones and acids (Owens et al., 1997).
Compounds such as dimethylsulfide (baked/roasted),
2-methoxyphenol (smoke, burnt), nonanal (fatty),
decanal (soapy, waxy) and 3-octanone (mushroom)
impart aromas which could certainly contribute to the
flavour of foods, if present at adequate concentrations
in the final product.
During fermentation, post-fermentation processing
and in-home cooking applications, it is likely that
several volatile aroma molecules are generated which
reflect the characteristic flavour for which dawadawa
is known. Evidence for the participation of indigenous
enzymes and flora in the development of the flavour
of dawadawa was presented by Ikenebomeh et al.
(1986). These authors demonstrated that both autoclaved
(sterile) and gamma irradiated (destroyed
indigenous flora) beans were unable to develop the
characteristic aroma of dawadawa. Additionally, the
M. Beaumont / International Journal of Food Microbiology 190 75 (2002) 189–196
typical pH increase observed in dawadawa fermentation
was absent; implicating that active microbial
metabolism is required in order to bring about the
changes observed in locust beans during fermentation.
2. Microbiology and fermentation of dawadawa
Seeds of the African locust bean tree (P. biglobosa)
are the traditional raw materials for the production of
Nigerian dawadawa. Other raw materials such as soy,
(Ogbadu and Okagbue, 1988) and groundnut (Amadi
et al., 1999) have also been used. The fermentation
process (including bean preparation and post-fermentation
handling) has been reported to be associated
with several benefits by converting otherwise tough,
inedible seeds into a valuable, flavour-enhancing
condiment (Odunfa, 1986). These benefits include:
enhanced digestibility due to degradation of nondigestible
oligosaccharides (e.g., stachyose, raffinose);
decreased flatulence potential; increased vitamin content
in the form of thiamine and riboflavin; reduction
or elimination of phytic and oxalic acids; extended
shelf life due to drying, alkaline pH and post-fermentation
additions (e.g., salt); protein hydrolysis to
peptides and amino acids, providing free glutamate,
as well as other amino acids that could function
directly in taste, or eventually serve as precursors
for aroma active molecules.
A general flow chart for dawadawa production,
adapted from Odunfa (1986) and Steinkraus (1995) is
shown in Fig. 1. The main steps of the process
involve extensive boiling and dehulling of the beans,
followed by a second boiling to soften the beans. The
cooked beans are then spread on to perforated trays
and covered with cloth or leaves, which provide the
moist environment necessary for the fermentation
step. Removal of the cover (and concomitant change
in the environment) promotes termination of the
fermentation, after which spices and additives (e.g.,
salt) are incorporated into the hydrolysed material.
Subsequently, the fermented mass can be formed into
shapes for convenient handling. Sun drying facilitates
stabilisation of the final product through reduction of
moisture. Though anecdotal in nature, it was found in
discussions with several individuals familiar with the
use of dawadawa that a 40–60-g dawadawa ball
may be used by a single family for up to 2 weeks or
more, depending on personal tastes and quantities
used.
Dawadawa production as described by Odunfa
(1986) is a multi-step process which does not include
a formal inoculation step. Bacteria required for the
fermentation appear to be incidental to both the
process and raw materials. Indigenous flora is likely
carried over from fermentation to fermentation in
sieves, trays and bags, which are repetitively used in
dawadawa production. Ambient contamination of
Fig. 1. Flow chart steps in the production of dawadawa (adapted from Odunfa, 1986 and Steinkraus, 1995).
M. Beaumont / International Journal of Food Microbiology 75 (2002) 189–196 191
spores from the local building/environment may also
contribute to the fermentation microflora.
Several workers have characterised dawadawa
fermentation and the microorganisms associated with
it (Odunfa, 1981; Antai and Ibrahim, 1986; Odunfa
and Oyewole, 1986; Ogbadu and Okagbue, 1988;
Odunfa and Komolafe, 1989; Aderibigbe and Odunfa,
1990; Olasupo et al., 1996). It is likely that fermentation
begins when the softened, washed dehulled beans
are placed in perforated trays and covered. This cover
most likely provides a moisture trap and possibly
serves as a source of inoculum. Changes over the
course of the 2–4-day fermentation period include a
pH increase from near neutral to approximately 8.0
(Odunfa, 1986); temperature increase from 25 jC to
as high as 45 jC (Odunfa, 1986); moisture increase
from about 43% up to approximately 56% (Odunfa,
1986); a fivefold increase in free amino acid content
and an increase in anaerobic mesophilic plate counts
from ‘‘not detectable’’ to 3105/g (Odunfa, 1986).
Glutamate concentration also increases almost fivefold
to 11.9 mg/100 g dawadawa (Odunfa, 1985).
Organisms reported to be isolated and/or characterised
from dawadawa include Bacillus subtilis (Odunfa,
1981; Ogbadu and Okagbue, 1988; Ikenebomeh,
1989), B. pumilus (Ogbadu and Okagbue, 1988), B.
licheniformis (Ogbadu et al., 1990) and Staphylococcus
saprophyticus (Odunfa, 1981). It is the hydrolytic
activity of microorganisms like those associated with
dawadawa (e.g., Aderibigbe and Odunfa, 1988,
1990), which can be used as a tool for providing a
base raw material for the generation of meat-like
flavours.
3. Process flavour development
Well controlled thermal reactions of selected precursors
under targeted conditions of time, temperature,
pH, aw and total solids content can result in
the development of Maillard and lipid reaction products
concomitant with the development of aromas
reminiscent of meat. Typical raw materials in the
development of such ‘‘process flavours’’ can include
reducing sugars (e.g., glucose, xylose, ribose), amino
acids (e.g., glycine, lysine), lipids (e.g., lecithin,
chicken fat, beef fat), reactive sulphur precursors
(e.g., cysteine, methionine, thiamine) and catalytic
agents. Additionally, the use of complex, proteinbased
hydrolysates can serve as a key starting raw
material for process flavour development. These bases
can include yeasts, acid or enzymatic hydrolysates,
and fermented protein bases. Selection of raw materials
and precursors in combination with processing
conditions can promote the generation of aromas
having some specificity, e.g., ‘‘white meat,’’ ‘‘red
meat’’ and ‘‘grilled’’ aromas.
Flavours developed in this way can impart taste
and aroma enhancement and specificity in the applications
in which they are used. Process flavours can
be directly applied for the enhancement of meat and
fish flavour in freshly prepared soups, sauces and
stews. Additionally, they can be used in prepared
commercial products such as canned, frozen and dried
foods and in culinary products. Process flavours can
also augment and boost the flavour enhancing properties
of bouillon and ‘‘taste makers.’’ These bouillon
products are generally commercially available as soft
and hard cubes for single-serve use and as powders
and pastes. Such products enhance aroma and taste
and provide ‘‘body.’’ This latter property is often
associated with salts of glutamic acid (e.g. monosodium
glutamate) and can be accentuated by 5V
nucleotides (e.g., IMP, GMP). Moreover, colour (to
match target application) dosing strength, granularity
and handling properties, including visual aspects of
cubes, are all relevant properties for successful culinary
flavouring products, such as bouillon cubes.
Sourcing of raw materials for process flavour
development is influenced by logistical, technical
and political considerations. For instance, hydrolysed
plant proteins (HPP’s), yeasts and enzyme-based
hydrolysates can serve as rich sources of precursors
for process flavour development, but sometimes have
inherent defects. Yeast-notes, off-flavours, and bitterness
can diminish the overall quality of process
flavours using these bases. Local availability of raw
materials is beneficial in minimising shipping costs
and in potentially stimulating the regional economy.
4. A patented process using Bacillus spp. as a
hydrolytic agent
On the basis of considerations discussed thus far,
the development of a patented process (Heyland et al.,
M. Beaumont / International Journal of Food Microbiology 192 75 (2002) 189–196
1995) which exploits the catalytic nature of Bacillus
as characterised in dawadawa production, offered
several benefits and possibilities for development of
process flavours and associated products. The overall
features of the patent are outlined in Fig. 2. In
summary, a protein-rich raw material is cooked and
inoculated with B. subtilis or B. natto and allowed to
undergo fermentation; the fermented base thus obtained
is mixed with water, reducing sugars and other
precursors and allowed to react at high temperature,
following which it is dried to a paste or powder. This
meaty flavour base, free of bitterness, is used as a
process flavour for a variety of applications and may
additionally contain an aroma reminiscent of dawadawa.
Several alternative processes and raw materials are
offered as extensions of this basic framework (Table
1). Raw materials may include cooked soy or carob
seeds, a variety of animal proteins, dairy proteins and
combinations of these raw materials. Pre-fermentation
additions—to augment the carbon source—may
include rice flour, barley malt and sugars. Pre-fermentation
processing of beans (e.g., boiling, grinding,
soaking) can facilitate the fermentation by increasing
the surface area and mass transfer of nutrients, and by
reducing competing microflora. Microbial inoculants
can originate from isolates identified in authentic
dawadawa, from Japanese natto, or from commercial
culture collections. Strains that exhibit limited mucilage
production, strong hydrolytic activity and have
the ability to generate significant quantities of glutamic
acid are preferred.
Variations can target additives and manipulations
after the fermentation process, in order to guide
process flavour formation, facilitate drying, or otherwise
to directly enhance aroma and taste properties.
These could include the addition of various sulphur
sources (cysteine, methionine, thiamine), sugars,
monosodium glutamate and sodium chloride. Finally,
options for thermal generation of the flavour, such as
extrusion or post-reaction drying with a vacuum oven
can also be employed.
One of the advantages of this patented process is
the ability to produce process flavours having a meaty
taste, which can also have a characteristic ‘‘dawaadawa-
like’’ or natto aroma, depending on the Bacillus
strain used in the fermentation. Studies conducted
in support of this patent demonstrated a correlation
between dawadawa aroma and the presence of 2-
methyl butanoic acid and 3-methyl butanoic acid in
the finished fermented protein base. However, since
this aroma is undesirable in some applications, it was
discovered that the aroma could be reduced by the
addition of assimilable carbohydrate sources prior to
fermentation. A parallel decrease in the levels of 2-
methyl butanoic and 3-methyl butanoic acid was
observed. This unexpected benefit allows the generation
of more neutral tasting meat-like process fla-
Fig. 2. Schematic outline of some points outlined in patent US 5,476,773 (Heyland et al., 1995).
M. Beaumont / International Journal of Food Microbiology 75 (2002) 189–196 193
vours without a strong characteristic dawadawa or
natto aroma.
Further benefits of this process include simplification
and standardisation of the fermentation process
for dawadawa production, the ability to use several
alternative local raw materials, generation of a nonbitter
natural flavour base and variations in dawadawa
or natto flavour depending on the nature of the carbohydrate
added at the pre-fermentation step. Moreover,
the dry format flavour obtained can be directly used,
or is applicable as an additive in culinary products
such as bouillon cubes.
5. Industrialisation of fermentation processes
At least three potential avenues for product development
from traditional fermentations are illustrated
in Fig. 3. Many commercial products have been
developed using the characterisation of food fermentations
as a basis. As an example, the current industrialisation
of several enzymes used in the food
industry is based on the characterisation of many
traditional processes in which animals, plants and/or
microbes were employed (Godfrey, 1998). In contrast,
the production of authentic fermented products on a
commercial scale would likely include such products
as Asian fish sauces and natto. Pure culinary and
functional targets would possibly include such products
as yogurts and certain soy sauces, where the
products impart performance and/or flavour characteristics
as primary targets. In a similar fashion, the
industrial manufacture of selected food ingredients
(e.g., amino acids, nucleotides, monosodium glutamate)
has also found roots in the technology of
traditional food fermentation (Steinkraus, 1982). The
flavour base derived from the Nestle patented process
is not intended to replicate the original dawadawa
product, but rather utilises enzymatic capabilities first
characterised in dawadawa to produce a more universal
neutral tasting meat flavour.
Technical parameters contributes to only one part
of the equation in the successful commercialisation of
fermentation products or processes. Where and how
these new products fit into a given market will largely
define business success if and when such processes
are industrialised. Understanding target populations,
the competitive environment, consumer expectations
and how new flavour products compete with existing
products are critical considerations. Some of this
information may only be acquired with consumerrelated
research, including focus groups, preference
panels, as well as descriptive sensory analysis. An
underlying element of success is defining what differentiates
the product offered from alternatives already
available in the marketplace. This in turn will of
course shape pricing, positioning in the consumer
environment, and how the product is promoted.
Fig. 3. Opportunities for industrial development from traditional
fermentations.
Table 1
Some alternatives elaborated in patent US 5,476,773 and corresponding
patents (Heyland et al., 1995)
Component Prefermentation Post-fermentation
Raw materials Pulse seeds Sodium chloride
Vegetable proteins
Animal proteins Sulphur sources:
Lactic proteins e.g., cysteine,
methionine,
thiamine
Rice Flour
Barley Malt Sucrose
Direct additions
carbon source,
e.g., glucose
Monosodium
glutamate
Process Boiling Heat reaction
Cooking Extrusion
Soaking Drying
Grinding
Microorganism source Dawadawa
Natto –
ATCC Culture
collection
M. Beaumont / International Journal of Food Microbiology 194 75 (2002) 189–196
Technical considerations will also weigh heavily
to determine if, where and how fermentation processes
are industrialised. Issues with forecasting
volumes, which in turn will influence the predicted
manufacturing capacity, require advance planning.
Availability of labour, competent suppliers and consistent
supply of raw materials, as well as basic
infrastructure, services and utilities are all necessary
for considerations for industrialisation. The cost of
maintenance and availability of replacement parts are
also valid considerations. An array of administrative
and political factors such as stability of local government,
currency exchange rates, cost of capital, legislation
and trade issues, import/export restrictions and
the current level of brand visibility of a company
will all add to a complete picture for industrialisation.
Commercialisation of small-scale fermentation can
bring clear benefits. These would include: standardisation
of products based on established manufacturing
targets; differentiation of products to conform to a
targeted consumer need or demand; product distribution
and availability; stimulation of the local economy
including employment, training, tax revenues and
enhanced use of local utilities and raw material
suppliers; safety and environmental standards; and
consumer education.
6. Opportunities
The use of traditional fermentations as a foundation
for commercial developments will likely continue and
accelerate in an era in which discoveries are augmented
by new technologies for understanding enzymes,
microorganisms and food flavours. Molecular biology
and affiliated technologies (e.g., PCR) can allowmicrobial
identification, detection of pre-selected marker
enzymes/genes and rapid sequencing and cloning from
candidate organisms. This can contribute to a more indepth
and rapid understanding of the organisms, their
roles, succession and prevalence in fermented foods.
Biochemical screening tools will continue to evolve
and allow rapid isolation and characterisation of new
enzymes.
Developments in the recovery and analysis of
aroma compounds from food matrices will also aid
in exploiting traditional fermentation processes (Peppard,
1999). A battery of techniques, including supercritical
fluid extraction and solid phase micro-extraction
(SPME) among other methods, can be used
to recover aroma active volatile molecules from food
products for subsequent analysis. Sensory linked
instrumental analysis using techniques such as gas
chromatography/olfactometry (GC-O) and aroma
extract dilution analysis can help define the role of
suspect aroma active molecules in complex mixtures.
Systematic application of such tools would not only
allow the discovery of new aroma compounds, but
would also allow enhanced definition of how these
contribute to flavour. The information obtained would
not only be applicable in targeting authentic fermented
food flavours, but would also be useful in
creating concentrated flavour ‘‘blocks’’ for use in
flavour blends.
In the case of dawadawa and related fermented
products where Bacillus spp. are implicated (e.g.,
natto, kinema, thua-nao), the interplay of microorganisms,
raw materials and process conditions can lead to
profound differences in the flavour and texture of
these products. Here lies a significant challenge in
understanding the underlying biochemistry and microbiology
leading to these differences. As a whole, it is
likely that knowledge gained by such efforts will be
used in either of two ways in the future: (a) safer,
more effective and consistent traditional fermentations,
which are still a viable part of the culture and
economy in their countries of origin; and (b) an
enhanced understanding of fermentations for more
effective exploitation of their constituent enzymes,
microbes and aroma molecules.
Acknowledgements
The author wishes to thank all those who
contributed to the preparation of this manuscript by
way of providing information, discussions and
critique. Special thanks are extended to the co-authors
of the patent, in particular S. Heyland, Nestle Product
Technology Centre (PTC) Kemptthal, Switzerland and
R. Wood, Nestle PTC Orbe, Switzerland. The input
and comments of Jens Van der Pol, Nestle PTC
Kemptthal, Switzerland and U. Zuercher, Nestle PTC
Orbe, Switzerland in preparing the oral presentation
are also appreciated.
M. Beaumont / International Journal of Food Microbiology 75 (2002) 189–196 195
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