Advances in Gluten-Free Bread Technology

Food Science and Technology
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Advances in gluten-free bread technology

PH Nitcheu Ngemakwe, M Le Roes-Hill and VA Jideani
Food Science and Technology International published online 16 May 2014
DOI: 10.1177/1082013214531425
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Article
Advances in gluten-free bread technology
PH Nitcheu Ngemakwe1, M Le Roes-Hill2 and VA Jideani1
Abstract
The unattractive appearance of gluten-free bread still remains a challenge in gluten-free breadmaking. In
response to this, additives such as dairy products, soya and eggs have been used to improve the quality of
gluten-free bread, but with limited success. In recent years, enzymes (transglutaminase and cyclodextrinase)
and hydrocolloids (carboxymethylcellulose and hydroxypropylmethylcellulose) have become the main focus
for the improvement of gluten-free bread. Transglutaminase has been shown to improve the dough visco-
elasticity and decrease crumb hardness (6.84–5.73 N) of the resulting bread. Cyclodextrinase also enhances
dough viscoelasticity, resulting in an improvement of 53% in shape index and crumb firmness. Similarly,
hydroxypropylmethylcellulose improves gas retention and water absorption of dough and reduces crumb
hardening rate of the resulting bread, while carboxymethylcellulose significantly increases dough elasticity
(60–70 BU) and bread volume (230–267 cm3/100 g bread).
Keywords
Gluten-free products, enzymes, hydrocolloids, functional properties
Date received: 2 January 2014; accepted: 11 March 2014
gluten is also important for crumb structure of wheat

INTRODUCTION derived products (Demirkesen et al., 2010).
Bread is one of the most common staple foods con- Non-wheat flours are typically characterized by their
sumed in the world. It is traditionally made with flour lack of gluten. The resulting dough does not have the
derived from the cereal, wheat. Other types of cereals, cohesive and elastic properties, because of the absence
pulses and legumes can be milled to give flour. of gluten (Lazaridou and Biliaderis, 2009). Hence,
However, the capability of the wheat proteins (gluten) gluten-free dough are more fluid than dough prepared
to transform wheat flour and water into a glutinous from wheat flour and are closer to cake batters in their
mass is currently limited to wheat, which upon viscosity and rheological properties (Cauvain, 1998).
baking, becomes bread (Cauvain and Young, 2007). This leads to poor dough formation with relaxing,
Gluten is important to obtain the desired volume and stretchable properties with limited machinability for
texture in a dough system through its role in gas reten- baking, and creating handling problems during
tion. The development of a strong protein network mixing. Consequently, the baked bread has a crumbling
required for the viscoelasticity and good dough rhe- texture, poor colour and other post-baking defects
ology is therefore essential. Glutenin and prolamin or (Torbica et al., 2010). However, gluten-free flours are
gliadin are the major fractions in gluten. Prolamin is a new substitute for wheat flour in the preparation of
responsible for viscosity and extensibility in a dough products consumed by wheat-intolerant individuals
system whereas glutenin develops dough elasticity and
cohesiveness (Cauvain and Young, 2007; Gujral and 1
Department of Food Technology, Faculty of Applied Sciences,
Rosell, 2004). Beyond improving the bread appearance, Cape Peninsula University of Technology, Bellville, South Africa
2
Biocatalysis and Technical Biology Research Group, Cape
Food Science and Technology International 0(0) 1–21
Peninsula University of Technology, Bellville, South Africa
! The Author(s) 2014 Reprints and permissions: Corresponding author:
sagepub.co.uk/journalsPermissions.nav PH Nitcheu Ngemakwe, Department of Food Technology, Faculty
DOI: 10.1177/1082013214531425 of Applied Sciences, Cape Peninsula University of Technology,
fst.sagepub.com P.O. box 1906, Bellville 7535, South Africa.
Email: nitcheupatrick@yahoo.fr

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Food Science and Technology International 0(0)
suffering from coeliac’s disease. Being a digestive dis- (Brown, 1993; Mani et al., 1992; Wang et al., 2004) for
order, coeliac’s disease is characterized by the damage starch degradation and sugar production. Water
of the villi, tiny hair-like projections in the small intes- hydrates the protein fractions which also assist in the
tine that absorb nutrients. This damage is due to an development of dough viscoelasticity (Chieh, 2006)
immunological reaction to gluten. According to and affects starch gelatinization during baking (Brown,
Demirkesen et al. (2010), coeliac patients do not toler- 1993; Mani et al., 1992; Wang et al., 2004). The amount
ate the gliadin fraction of wheat and the prolamins of water added during dough mixing mainly depends on
absorption of certain cereals such as rye and barley. water absorption of the gluten-free flour. Thus, water
The only treatment of this disease is for people who absorption increases with increase in protein content
suffer from coeliac’s to adhere to a strict gluten-free and with increase in flour extraction. Water binds flour
diet throughout their lifespan (Demirkesen et al., and other dough ingredients into a coherent mass and
2010). Therefore, the interest in alternative gluten-free dissolves certain ingredients for development of yeast
grains such as oats, rice and sorghum is increasing due and for leavening action at the baking stage. Water add-
to the consumer demand for novel and healthy foods ition reduces the viscosity and increases dough extensi-
that are gluten free. Attempts have been made by sci- bility. If the water volume is too low, the dough becomes
entists to manufacture gluten-free bread products. Our brittle, not consistent and highlights a marked ‘crust’
objective was to review recent advances in gluten-free effect due to the rapid hydration. Water content and
bread technology as a comprehensive guide for infor- its distribution therefore play an important role in tex-
mation on the subject. tural properties of bread such as softness of crumb,
crispness of the crust and shelf-life.
TECHNOLOGY OF GLUTEN-FREE
BREAD PRODUCTION Salt
The most basic gluten-free bread ingredients consist of Salt is added at about 1.5% of gluten-free flour weight
gluten-free flour, water, yeast and salt. The sugar add- for taste (Chieh, 2006; Gray and BeMiller, 2003;
ition is not essential since flour amylases convert starch Lemmer, 2009; Zobel and Kulp, 1996) and to improve
to sugars. These sugars are metabolized by yeast for gas dough handling. Salt slows down water imbibition and
production. Nevertheless, even those most skilled in the swelling of flour proteins; reduces dough extensibility
art of baking agree that at the very least it is difficult to and improves gas retention, bread crumb and slicing
make bread of a high, consistent quality from only properties.
these raw materials. The baker always adds small
amounts of extra ingredients to enhance dough per-
Yeast
formance during processing or to improve the quality
of the finished product. The principal benefit is related Saccharomyces cerevisiae added at about a 2% concen-
to the properties of the final baked product and the tration (gluten-free flour basis, weight/weight) is used in
modification of the dough during processing. Besides the baking industry (Williams and Pullen, 1998; Zobel
these ingredients psyllium husks, dairy products (yog- and Kulp, 1996) for its ability to produce gas through
hurt), soya, eggs and pea proteins are commonly used the metabolism of glucose. Yeast ferments glucose to
as gluten replacers in gluten-free bread formulations. produce carbon dioxide and ethanol under anaerobic
Each ingredient has a specific role in the baking of conditions (Gray and BeMiller, 2003; Williams and
gluten-free bread. The functionalities of the ingredients Pullen, 1998; Zobel and Kulp, 1996). Brown (1993)
and production processes for gluten-free breadmaking reveals that the carbon dioxide goes into the dough/
are the main topic of discussion in this section. water phase when it becomes saturated and is released
into a gas cell that is formed during dough mixing.
Yeast also contributes to the flavour of baked products
Water
by the fermentation by-products produced (Lemmer,
Water influences the dough consistency, dough rhe- 2009) through releasing reducing sugars that react
ology and dough temperature (Brown, 1993; Wang with the amino groups of proteins during baking.
et al., 2004). It hydrates proteins and carbohydrates S. cerevisaie is considered to be amongst one of the
for the development of dough and hence acts as a dis- major yeasts used in dough fermentation and has an
persing agent bringing the ingredients into contact with important effect on dough rheological properties.
each other and dissolving the soluble ingredients Research has shown that the effect of the yeast on rheo-
(Brown, 1993; Chieh, 2006; Mani et al., 1992; Wang logical properties is similar to the effect of hydrogen
et al., 2004). Dough is formed when mixing water and peroxide (Mirsaeedghazi et al., 2008). This fact indi-
flour, resulting in activating enzymes such as amylases cates that the effect of yeast on rheological properties
2
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Nitcheu Ngemakwe et al.
is due to the production of hydrogen peroxide by the et al., 2007) usually due to the loss of intermolecular
yeast. The carbon dioxide produced during fermenta- attractions between ingredients responsible for crumb
tion dissolves in water, resulting in a decrease in pH. formation and is usually associated with the loss of
Hence, carbon dioxide affects rheological properties of water (Gomez et al., 2007). Water migrates from
the fermented dough such as gluten-free dough (Spies, crumb to crust during staling and leads to a glass-to-
1997). Furthermore, Salvador et al. (2006) demon- rubber transition of the two components. Zobel and
strated that dough samples containing yeast show Kulp (1996) described the mechanism of starch retro-
lower elastic, viscous and viscoelastic moduli than the gradation leading to staling. Due to the formation of
control sample and greater frequency dependence, par- double helical structures and crystalline regions, the
ticularly at the higher frequencies, within the period migration of water and amylopectin retrogradation is
studied. However, Wehrle and Arendt (1998) reported considered to be primarily the cause of bread staling
that yeasted dough had a lower recovery capacity. In during ageing (Gray and BeMiller, 2003; Zobel and
other words, they were less elastic than unfermented Kulp, 1996). Changes in the firming rate of bread are
dough. Yeast at 2 and 4% concentrations (weight/ due to hydrogen bonding between protein and starch
weight) showed very similar behaviour but the 8% granules, where protein is cross-linked by gelatinized
yeast sample presented the lowest viscoelastic constant starch (Martin et al., 1991). In one study, bread
values. This behaviour and the lower moduli values baked from flours with a low (10.4%) protein content
indicated a weaker, less structured gel with a more staled at a faster rate than those baked from flours with
viscous-like behaviour. In all the yeast dough samples, a higher (13.1%) protein content. Maleki et al. (1980)
the viscoelastic moduli showed a similar behaviour in concluded that the gluten-free flour component primar-
relation to the temperature. The presence of yeast does ily responsible for the shelf-life of bakery products is,
not appear to induce delays in the gelatinization onset therefore, protein. Acting as a diluent, protein slows the
temperatures. staling rate of starch (Pateras, 1998; Zobel and Kulp,
1996). However, Gray and BeMiller (2003) and Kestin
et al. (2004) suggested that the starch–protein inter-
Gluten-free flour
action is responsible for the firming process because
Gluten-free flour such as fonio, rice, rye, maize, oats, swollen starch and protein cross-link during baking
buckwheat, teff and blends consists of various compo- (Kamel and Ponte, 1993; Kestin et al., 2004). The kin-
nents, namely protein, starch and minerals (Sluimer, etic energy of crumb decreases during staling which
2005; Sultan, 1990). The starch and protein compo- allows cross-linkages to increase both in number and
nents are essential because they are important for the in strength thus resulting in the firming of the crumb
transformation of a gluten-free dough foam-type (Kamel and Ponte, 1993; Kestin et al., 2004). Hence,
system to a bread-like system (Hug-Iten et al., 1999). Martin et al. (1991) proposed a model of bread staling
Starch is relatively inert during dough mixing, but con- that incorporates the role of starch and protein. They
tributes to increased dough viscoelasticity through its argued that bread firming results from interaction
filling function. Starch is situated in spherical granules between the continuous protein matrix and discontinu-
(Karim et al., 2000; Pateras, 1998) and is made up of ous remnant of starch granules. Poor quality flour has
two polymers, namely amylose and amylopectin. more hydrophilic properties than good quality flour
Amylose is a linear polymer and a determining key (He and Hoseney, 1991; Martin et al., 1991).
factor for initial loaf volume whereas amylopectin is a Consequently, poor quality protein interacts more
branched polymer (Blanshard, 1986; Karim et al., 2000; strongly with starch granules in dough. These inter-
Pateras, 1998). Amylopectin is responsible for crystal- actions are stronger during and after baking, increasing
linity, while amylose is in a more amorphous state the tendency of bread firming.
(Blanshard, 1986; Karim et al., 2000; Pateras, 1998). It is known that cereal flours other than wheat do
The solubilized amylose forms a continuous network not contain gluten. Hence, certain ingredients have
during cooling by which swollen and deformed starch been used as gluten replacers. Such ingredients are
granules are embedded and interlinked. Consequently, discussed below.
bread loses its freshness during cooling and stales
(Eliasson and Larsson, 1993). The crust toughens and Psyllium husks. Psyllium husks or Ispaghula is a
the crumb becomes more firm and less elastic losing common name used for several members of the plant
moisture and flavour (Hoseney, 1994). Starch retro- genus Plantago whose seeds are used commercially for
gradation involves the re-association of starch compo- the production of mucilage. It develops ‘weak gel’ net-
nent molecules into a partially crystalline, ordered works which trap carbon dioxide generated during
structure (Ronda and Roos, 2011). As a result, the proofing and therefore increases gas retention and
ageing of bread diminishes cohesiveness (Gomez loaf volume (Zandonadi et al., 2009). It is stable at
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various pH levels and temperatures and it is similar to Sanderson, 1987; Kenny et al., 2000; Mannie and
gluten in food. Therefore, Zandonadi et al. (2009) sug- Asp, 1989). Dairy products such as yoghurt used in
gested that psyllium can replace gluten in recipes gluten-free bread formulas increase water absorption
because of its contribution in developing dough and, therefore, enhance the handling properties of the
viscoelasticity. dough (Gallagher et al., 2004a; Nunes et al., 2009).
Considered as fibre, psyllium does not modify the However, Houben et al. (2012) showed that the main
dough development time (DDT) or the stability. whey proteins contained in yoghurt, the a-lactalbumin
However, Laurikainen et al. (1998) reported an increase (four disulphide bonds) and the ß-lactoglobulin, which
in DDT and a decrease in stability with 5% rye bran. can be a monomer, dimer and an oligomer depending
Greater effects are observed on the mixing tolerance on pH value, ionic strength and temperature, have a
time (MTT) which is the difference in Brabender globular structure and hydrophobic, compact folded
Units between the top of the curve at the peak and polypeptide chain. Hence, decreases water absorption
the top of the curve measured 5 min after the peak is of dough.
reached and elasticity. Both MTT and elasticity are Dough viscosity is one of the key elements which
reduced by the addition of fibres. The extent of the determine the rheological quality of gluten-free bread.
decrease depends on the type of fibre. These results Gallagher et al. (2003) showed that increasing water
can be explained by the interactions between fibres addition in the dough resulted in increased loaf vol-
and protein, as described by Chen et al. (1988). umes. These findings are confirmed by Schober et al.
The addition of fibre to gluten-free flour modifies the (2005) in their study on gluten-free dough made from
rheological properties of the dough to a lesser extent sorghum flour. Dairy proteins possess functional prop-
compared to bran (Wang et al., 2002), increases the erties similar to gluten, as they are able to form net-
configuration curve ratio (P/L) (P ¼ tenacity or resist- works and have good swelling properties (Gallagher,
ance to extension, L ¼ dough extensibility), improves 2009). However, dough with added dairy product dis-
proofing stability and increases dough stability plays a much higher ability to resist deformation and
(Annon, 1999). Fibre in gluten-free dough interacts led to greater solid-like behaviour under the applied
with the proteins resulting in increased dough resistance testing conditions (Bertolini et al., 2005). This might
to deformation or tenacity (P) (Wang et al., 2002). be mainly due to the presence of proteins such as
Dietary fibre such as psyllium husks is the edible milk protein isolates and sodium caseinate in the
portion of plants (or analogous carbohydrates). It is dairy product, decreasing water absorption of dough
resistant to digestion and absorption in the human (Bertolini et al., 2005). This finding nicely correlates
small intestine with complete or partial fermentation with studies performed by Nunes et al. (2009) on rice
in the large intestine (Gelroth and Ranhotra, 2001). flour. Bertolini et al. (2005) also showed that the stor-
From the technological view, fibre incorporation age modulus (G0 ) increased when sodium caseinate was
improves the texture, sensory characteristics and present in the composite flours. This effect seemed to be
shelf-life of foods due to their water binding capacity, clear, mainly for rice starch, at the lower starch concen-
gel forming ability, fat mimetic, texturizing and tration, suggesting that the viscoelastic properties of the
thickening effects (Gelroth and Ranhotra, 2001; sodium caseinate are more important in the systems
Thebaudin et al., 1997). The addition of dietary fibre with low starch concentration. Despite the conflicting
in gluten-free formulations improves gas retention abil- results, it seems clear that changes in the viscoelastic
ity yielding breads with significantly higher loaf volume properties of the systems could be attributed to the
and crumb softness compared to the control limitation of starch swelling and gelatinization by
(Zandonadi et al., 2009). The gluten-free breads with sodium caseinate (Bertolini et al., 2005). The effect of
dietary fibre provide the consumer with higher amounts sodium caseinate on starch swelling was indicated by
of total dietary fibre and have an appealing dark crust water restricted in the system and is more evident in
and a uniform and finely grained crumb texture starches with high amylose content (Bertolini et al.,
(Sabanis et al., 2009). 2005).
In general, dairy powders with high protein/low lac-
Dairy products. The incorporation of dairy ingredients tose content (sodium caseinate, milk protein isolate)
is well established in the baking industry (Stahel, 1983; give breads with an improved overall shape and
Zadow and Hardham, 1981). Dairy proteins are signifi- volume (Gallagher et al., 2003) and a firmer crumb tex-
cantly functional ingredients and due to their versatility ture (Arendt et al., 2008). When optimal water is added
can be readily incorporated into many food products. to the gluten-free formulation these breads exhibit
They are used in bakery products for both nutritional increased volume and a much softer crust and crumb
and functional benefits including flavour and texture texture than the controls. Supplementing the gluten-
enhancement, and storage improvement (Cocup and free formulation with high protein-content dairy
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powders doubles the protein content of the breads promote significant decreases of 14 and 61% of the
(Gallagher et al., 2004a). Whole and skimmed milk final viscosity and setback, respectively, when com-
powders improve sensory characteristics. Sodium case- pared to the control of rice dough (Marco and Rosell,
inate and hydrolysed casein display beneficial func- 2008a). However, it significantly increases by 69% the
tional properties in breadmaking including low proof storage or elastic modulus (G0 ) of rice dough (Marco
time, high volume and low firmness (Kenny et al., and Rosell, 2008a). As reported by Marco and Rosell
2000). Therefore, breads containing dairy ingredients (2008a), the same tendency is noted on viscous modulus
exhibit the best quality and resemble a wheat bread (G00 ) with independent frequency. The increase of
most closely (Gallagher et al., 2003). The high quality dough viscosity in the presence of soya proteins could
of bread that contains dairy ingredients is attributed to be attributed to its higher water binding capability.
the capacity of these ingredients to form a network The presence of soya proteins hardly modifies the
similar to gluten. emulsifying properties of rice dough. Marco and
The colour analysis of gluten-free bread shows that Rosell (2008a) pointed out that the incorporation of
the bread supplemented with dairy products has lower soya proteins increases the emulsifying properties of
crust L* (value given a darkness to lightness indicator rice dough. It may be due to the process followed
for products) values than the control (Gallagher et al., while it is produced, since this process can impact the
2003). This is due to the small amount of lactose con- solubility and the degree of hydrophobicity, modifying
tained in dairy powder which is involved in Maillard the emulsion properties of the proteins (Petruccelli and
browning and caramelization reactions (Gallagher Anon, 1994).
et al., 2003). These reactions are affected by the distri- Supplementing gluten-free formulations with soya
bution of water and the reaction of reducing sugars and proteins leads to the improvement of the quantification
amino acids resulting in a darker crust colour of free amino groups of the resulting products. As is
(Gallagher et al., 2003). However, crumb colour is expected its incorporation results in an increase in the
not significantly affected by dairy product (Gallagher amount of the free amino groups, as the result of the
et al., 2003). Gluten-free bread supplemented with dairy increase in the protein content (Marco and Rosell,
products has an appealing dark crust and white crumb 2008a). This increase can be explained by the increase
appearance (Gallagher et al., 2004b). in solubility of the soya proteins resulting from the
The incorporation of dairy products in gluten-free deamidation reaction (Babiker, 2000).
bread renders it ‘looking more like real bread’ Soya proteins addition in gluten-free products
(Gallagher et al., 2003). The crumb is ‘more even and improves crumb grain score, bread volume and overall
more airy than the control’ and the loaves have ‘better bread score (Sanchez et al., 2002) and increases water
volume and crust colour, like wheat bread’ (Gallagher absorption of gluten-free dough. Similar findings are
et al., 2003). As a result, most of the panellists show shown by Houben et al. (2012) on gluten-free flours.
interest in bread supplemented with dairy products
than the control (Gallagher et al., 2003) leading to Eggs. Egg proteins such as albumins and globumins
good acceptability scores in sensory tests (Gallagher are able to form strong, cohesive and viscoelastic
et al., 2004a). films essential for stable foaming (Moore et al., 2007).
They form viscous solutions, a film-like continuous
Soya. Soybeans belong to the Fabaceae family, which protein structure similar to that of wheat gluten.
contains legumes or pulses. Soybeans have a number of Using as gluten replacer in the baking, egg proteins
properties that renders them an attractive ingredient for mainly act as a foaming agent, as a crumb stabilizer
functional foods. Soya has positive impacts on bone and for creating a good shape because of their border
tissue and hence reduces osteoporosis, the risk of car- areas activity.
diovascular disease and prevents breast cancer. It is The incorporation of egg proteins improves the past-
used to increase the protein content and the structural ing properties of gluten-free dough. Egg albumin sig-
properties of gluten-free products. Soya protein shows nificantly decreases by 34% the breakdown of rice
strong gel-forming properties and is used for the pro- dough compared to the control and significantly
duction of emulsions and foams. Its functionality increases the viscosity of gluten-free dough (Marco
depends on the environmental factors such as pH and Rosell, 2008a). Egg albumin proteins exhibit a
value, ionic strength and temperature. The soya pro- slight increase in elastic modulus (G0 ) and viscous
teins are classified into two groups, globulins (repre- modulus (G00 ) with frequency (Marco and Rosell,
senting 90% of the total amount) and albumin 2008a). The swelling ability of the egg albumen proteins
(representing 10% of the total amount). in gluten-free dough leads to a viscous fluid showing a
The addition of soya in gluten-free formulations similar network protein structure function than the one
improves the pasting profile of dough. Soya proteins known from gluten (Houben et al., 2012).
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The presence of egg albumin increased the 2008b; Mariotti et al., 2009). This increase in water
emulsifying properties of rice flour (Marco and absorption might be attributed to its water holding cap-
Rosell, 2008a). These emulsifying properties are acity, which is about 2.7–2.8 g/g for pea protein (Marco
induced by the heat coagulation of the protein and and Rosell, 2008b).
the egg yolk containing phospholipids and lipoproteins. The incorporation of pea protein significantly
As a result, they facilitate the dispersion and stabiliza- impacts the viscoelasticity properties of gluten-free for-
tion of gas bubbles in the gluten-free dough systems mulations. According to Marco and Rosell (2008b), the
(Houben et al., 2012) because egg white proteins increase in the amount of pea protein produces a sig-
containing ovalbumin (54%) include four thiol and nificant linear decrease in the storage or elastic (G0 ) and
one disulphide group able to stabilize the gel by viscous modulus (G00 ) of rice dough derived from a sig-
polymerization via the thiol disulphide exchange. nificant quadratic effect. The increase in G0 and G00
The egg yolk is often used as emulsifier in the baking could be mainly due to the high water holding capacity
industry because of its coagulation power during of pea protein, since constant water absorption is used
the thermal treatment creating similar thermo- (Marco and Rosell, 2008a). Findings presented regard-
irreversible gels. ing the effect of pea protein on the water absorption
The incorporation of egg albumin proteins confirmed this assumption. However, Marco and
modifies the quantification of free amino groups of Rosell (2008a) noted a significant increase in G00 and
gluten-free formulations. It results in an increase in G0 of rice dough while adding 5% of pea protein. In
the amount of the free amino groups, increasing the addition, Mariotti et al. (2009) demonstrated that the
protein content of gluten-free product (Marco and higher amount of pea protein decreases the elastic
Rosell, 2008a). As soya proteins, this increase might modulus (G0 ) and its impact is more evident on G0 at
be also attributed to the increase in solubility of the temperatures lower than the starch gelatinization tem-
egg albumin proteins resulting from the deamidation peratures. The final viscosity of rice dough is decreased
reaction (Babiker, 2000). by 20% while adding pea protein (Marco and Rosell,
Eggs improve the rheological properties of gluten- 2008a).
free bread. They increase bread volume and the As a good indicator, rheological assessment is a
amount of pores per square centimetre (Moore et al., good tool to evaluate the polymer molecular structure
2006). Egg albumin increases the gas-binding capacity and the end-use performance of gluten-free products.
by connecting the starch granules (Jonagh et al., 1968). The level of pea protein significantly affects gluten-free
According to Houben et al. (2012), the best crumb tex- dough hardness (Marco and Rosell, 2008b). The
ture of gluten-free bread is reached by the addition increase of the amount of pea protein increases the
of full egg powder compared to the other protein resulting dough harness (Marco and Rosell, 2008b).
sources. The increase in interface of liquid and air is However, pea protein content does not significantly
induced by the denaturation and aggregation of egg affect the dough cohesiveness (Marco and Rosell,
white inside the ovalbumin during the pitching. Stable 2008b). The supplementation of gluten-free formula-
gas foam is formed, which loses the dough and stabil- tions by pea protein leads to dough with higher springi-
izes the dispensation of further ingredients (Houben ness when protein level increases and results in a
et al., 2012). The fibrillar structures of gas bubbles positive linear effect on gumminess. Marco and Rosell
are stabilized by the egg white protein ovomucin. (2008b) pointed out that stickiness shows a quadratic
Therefore, the protein network reduces the swelling positive dependence on the incorporation of pea
and gel forming of the starch leading to the coagulation protein.
and prevention of the coincidence of the bread during The presence of 5% of pea protein hardly
baking. modifies the emulsifying properties of rice dough. It
could be attributed to its hydration capacity since
Pea protein. Proteins from different sources such as water acts as a plasticizer improving the functional
pea protein are traditionally added in gluten-free properties of the dough (Marco and Rosell, 2008b).
products to increase its nutritional benefits because Thus, pea protein relatively possesses good emulsifying
gluten-free products after mixing have a very low properties with low emulsion stability compared to the
protein content and are lysine deficient (Marco and properties of soybean protein (Marco and Rosell,
Rosell, 2008a). However, its presence in gluten-free for- 2008a).
mulations impacts other properties. The microstructure analysis of rice dough containing
The addition of pea protein significantly affects the pea protein shows that pea proteins present aggregates
farinograph water absorption of gluten-free dough. The of distorted spherical structures (Marco and Rosell,
higher the amount of pea protein added, the higher the 2008b). Besides, when the higher amount of pea isolate
water absorption of rice dough is (Marco and Rosell, (6%) is added, as expected, more green area is
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proportionally seen, highlighting a larger protein counterparts (Table 1). The function of main steps in
matrix (Mariotti et al., 2009). gluten-free breadmaking process is described below.
The protein analysis of rice dough reveals that the
addition of pea protein isolate produces an increase in
Weighing and mixing
proteins extracted in the albumin–globulin fraction
(Marco et al., 2007). All the ingredients are weighed according to the defined
Therefore, the incorporation of pea protein in standard or formulation (including minor ingredients).
gluten-free formulations improves the experimental The ingredients are blended and hydrated with water in
gluten-free dough from both a physical (rheological order to develop dough and incorporate air bubbles.
and ultrastructural) and nutritional point of view, and Dough connotes a semi-solid mass that resists mixing.
a delay in the staling phenomenon of these samples Stauffer (1998) revealed that a typical mixogram shows
might be expected (Mariotti et al., 2009). the various stages of dough formation, namely hydra-
tion, blending and breakdown.
Optional ingredients
Hydration. In flour, most of the protein is considered as
The main optional ingredients used in the gluten-free flinty material. The mixer firstly hastens the conversion
baking industry are fat, sugar, emulsifiers, oxidizing of the flinty protein bodies into soft, hydrated (but not
agents, reducing agents, enzymes and fermentation truly dissolved) protein dispersions. The protein is fur-
accelerators (Cauvain, 1998). Fat makes the crumb ther modified during gluten development due to the
finer and silkier in small amounts (up to about 3%, absorption of water from the water-soluble flour com-
flour weight) and increases loaf volume and freshness ponents (and added water-soluble ingredients such as
retention. Sugar improves both fermentation and salt and sugar). When water is brought into contact
browning; improves dough stability, elasticity and with the flour particles and the process is observed
shortness; and makes the baked product somewhat under a microscope, the particles seem to explode;
mellow (Cauvain and Young, 2007). Emulsifiers can strands of protein are rapidly expelled into the aqueous
be dispersing agents (lecithin, hydroxylated lecithin), phase (Bernadin and Kasarda, 1973). Movement of the
volume improvers, dough strengtheners (polysorbate cover glass stretches the protein indicating their exten-
60) and/or crumb softeners (monoglycerides and digly- sibility (Amend and Belitz, 1990). The input of mech-
cerides). Oxidizing agents such as potassium iodate, cal- anical energy is important to dough formation. For
cium peroxide and calcium iodate oxidize protein. They instance, a thick slurry that has no dough-like proper-
enhance the gas retention of dough by strengthening ties when stirred increases in consistency, forming soft
the gluten and increasing absorption. Reducing agents (undeveloped) dough. Hydration alone is not sufficient
ensure the dispersal of proteins to reduce mixing for dough making.
requirements (e.g. L-cysteine). Enzymes such as amyl-
ases provide fermentable carbohydrates which stimu- Blending. Particles of flour are agglomerates of starch
late gas production in dough (diastatically active malt granules embedded in a network of protein. As the pro-
preparations). Fermentation accelerators (ammonium tein network is softened by hydration and agitated by
phosphate, ammonium sulphate, etc.) provide nitrogen mixing, the starch granules become less firmly attached
sources for yeast metabolism. to the protein but remain associated with the protein
fibres. Most of the starch is removed by washing and
GLUTEN-FREE BREADMAKING kneading the dough but it cannot be totally removed.
During this early stage of mixing, all the ingredients in
PROCESSES the dough are blended to give a homogeneous dough
All the processes almost include the same unit oper- mass. Lipids are uniformly distributed and brought
ations as described in Table 1. The production of into contact with the protein fibres, and soluble mater-
gluten-free breads is slightly different to that of stand- ials are fully dissolved and distributed in the aqueous
ard wheat breads in terms of the regulation of physical matrix.
parameters and the absence of gluten. As wheat dough,
gluten-free dough is traditionally mixed, bulk fer- Breakdown. Peak development is reached when the
mented, divided/moulded, proofed and finally baked. dough becomes softer and less resistant to mixing
However, several studies show that gluten-free dough action. During this peak development process, the
have the tendency to contain higher water levels and dough loses its ability to retain gases during proofing
tend to have a more fluid-like structure (Bernadin and (Cauvain, 1998). The viscosity of dough proteins
Kasarda, 1973). Therefore, they require shorter mixing, extracted into 1% sodium dodecyl sulphate solutions
proofing and baking times than their wheat is lowered in over mixed dough compared to optimally
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Table 1. Gluten-free bread production processes
Cereal flour Unit operation Authors
Fonio (acha) Dissolving sugar in 80% warm water; adding yeast Jideani et al. (2007)
and allowing to stand for 10 min, pre-mixing the
remaining sugar with other dry ingredients; mixing
of dry ingredients, water containing yeast mixture
using wooden spoon (5 min); first proofing in the
bowl for 30 min; moulding and panning; second
proofing (45–50 min); baking (215 C, 5 min; 180 C,
40–50 min); cooling (15 min); packaging in poly-
ethylene bag.
Wholegrain oat Dissolving baker’s yeast and sugar in a solution of Huttner and Arendt (2010)
water (30 C, 10 min); placing remaining dry ingre-
dients and activated baker’s yeast in a mixing bowl;
mixing all the ingredients (level 2 for 30 s and level 4
for 1.5 min); dough scaling (450 g) and panning;
proofing (30 C, 85% relative humidity, 30 min) in a
proofer; baking (190 C, 45 min); depanning; cool-
ing (2 h, room temperature).
Sorghum and pre-gelatinized Mixing all the ingredients (low gear, 10 min); dough Onyango et al. (2010)
cassava weighing (1200 g) and panning; proofing (33 C,
85% relative humidity, 55 min); baking (210 C,
35 min); depanning; cooling (2 h, 25 C); packaging
in polythene bag.
Rye Dissolving TG in water; mixing all the ingredients Beck et al. (2011)
(30 C, 60 s, 53 r/min; 120 s, 106 r/min); proofing
(60 min, 30 C, 80% relative humidity); baking
(60 min, 240 C).
Rice Dissolving yeast in warm water (35 C); adding yeast Lazaridou et al. (2007)
to dry ingredients and sunflower oil; mixing all the
ingredients (2 min, speed 3); proofing (25–30,
20 min); baking (215 C, 20 min); cooling (1 h, room
temperature); packaging in polypropylene bag.
Buckwheat þ rice þ corn þ Dissolving dried yeast and sugar in a solution of water Renzetti et al. (2008b)
oat þ sorghum þ teff (22–26 C); pre-fermenting the mixture in a proofer
(30 C, 85% relative humidity, 10 min); mixing all the
ingredients (2 min, speed 2 out of 6); dough scaling
(400 g) and panning; proofing (30 C, 85% relative
humidity, 30 min); baking (190 C, 35 min); depan-
ning.
Cooling (90 min, room temperature).
Rice þ buckwheat Mixing flours and salt (speed 1, 5 min, 30 C); dissol- Peressini et al. (2011)
ving yeast in a portion of water (30 C); mixing all
the ingredients (speed 2, 5 min); dough scaling
(250 g) and panning; proofing (30 C, 85% relative
humidity, 45 min); baking (200 C, 50 min).
mixed dough, indicating a smaller average molecular During the mixing, bulk fermentation occurs, which
weight (Danno and Hoseney, 1982). The breakdown produces flavour development and allows dough devel-
phenomenon is due to the presence of ferulic acid in opment. Punching expels gas and subdivides the exist-
the water soluble fraction of flour (Schroeder and ing gas cells, thereby incorporating air into the dough
Hoseney, 1978). Dough breakdown simply appears mass. The dough mass is divided according to the
to be a continuation of the process by which flour standard process defined, rounded by the shaping of
protein is converted to medium-length protein poly- the dough piece into a shape to allow proofing to
mers that impart the desired rheological properties occur. During the first proof, stresses in the dough
to dough. relax, resulting in improved handling properties.
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Prior to a final proofing process, the dough piece is Koppelman, 2002). In the absence of primary amines,
shaped into a cylindrical form and placed into bread water molecules are used as acylacceptors and the g-
pans. For the final proofing, the production of CO2 by carboxamide side chains are deamidated, forming glu-
the yeast allows the dough to rise while in the bread tamic acid residues (Motoki and Seguro, 1998). Table 2
pans. The exposure to heat during baking sets the loaf summarizes the effect of TG on the functional proper-
structure and develops the baked flavour and colour of ties of wheat and gluten-free dough.
the bread. The final baked product is removed from the Many studies with wheat flour show that TG has a
tins during the depanning step. Cooling allows for the dough-strengthening effect. Thus, TG increases the
slicing of the bread and prevents any moisture migra- extensibility and modifies the elasticity behaviours of
tion onto wrapping or packaging (cooled at a tempera- dough by improving the protein network (Basman
ture of 27 C in cold air, with a residence time of et al., 2002; Bauer et al., 2003; Caballero et al., 2007)
approximately 2 h). Bread loaves are sliced and and water-holding capacity, reducing therefore the
wrapped for hygienic, aesthetic and convenient presen- required work input during mixing (Basman et al.,
tation to the consumer (reciprocating frame of blades, 2002).
followed by automatic wrapping). The addition of TG at less than 1% significantly
increases the elastic (G0 ), viscous (G00 ) and viscoelasti-
USE OF ENZYMES AND city (jG*j) modulus values of buckwheat dough
HYDROCOLLOIDS IN GLUTEN-FREE (Kohajdova and Karovicova, 2009). This increase of
dough elasticity and viscosity suggests that TG leads
BREADMAKING TECHNOLOGY
to protein cross-linking and network formation of
Besides the ingredients used in wheat breadmaking, buckwheat dough, thereby modifying its viscoelasticity
psyllium husks, soya and eggs are commonly used as properties (Han et al., 2013). This modification is said
gluten replacers in gluten-free bread formulations to to be due to the formation of non-disulphide covalent
solve the above rheological challenges but with limited cross-links between peptide bound g-glutamyl residues
success. Recently, the effects of transglutaminase (TG) and "-amino groups of lysine residues in proteins (Shin
and cyclodextrinase (cyclodextrin glycosyl transferase, et al., 2010). Similar results have been reported by
CG) on gluten-free formulations have been studied Renzetti et al. (2008b) and Shin et al. (2010) in their
(Gujral et al., 2003; Huang et al., 2010). TG catalyses studies on rice dough. However, upon increasing the
cross-link formation while CG degrades starch. The TG amount in the buckwheat dough from 1.0 to
effects of hydrocolloids such as carboxymethylcellulose 1.5%, the values of jG*j and G00 were not significantly
(CMC) and hydroxypropylmethylcellulose (HPMC) in changed (Han et al., 2013). It is probably due to the
wheat-free formulations have also been documented restricted reactivity of the TG owing to sufficiency of
and have been found to modify the functional proper- lysine but rather the relative content of glutamine in the
ties of gluten-free bread products through their action buckwheat flour (Renzetti et al., 2008b). Huang et al.
as water binders. This section gives an overview on the (2010) reported that the mechanical spectrum of all oat
functional properties of these additives and their role in dough indicates the elastic modulus (G0 ) is always
gluten-free formulations. higher than the viscous modulus (G00 ), and both
increase with increasing levels of TG.
Consequently, TG leads to protein cross-linking and
Effect of TG on the functional properties of
the formation of a network structure which causes a
gluten-free dough and bread
modification in the viscoelastic properties of the oat
TG is a family of enzymes (EC 2.3.2.13), which cata- dough and other gluten-free dough. The viscous modu-
lyses reactions over a temperature range of 0–65 C lus (G00 ) shows a higher increase in amplitude than the
(with optimal temperature 50–55 C) and over a pH elastic modulus (G0 ). The highest viscoelastic dough is
range of 4–9 (with optimal pH 6–7). It catalyses an obtained when the enzyme is added at a final concen-
acyl-transfer reaction between the g-carboxamide tration of 1.0% (Huang et al., 2010), implying that TG
group of peptide-bound glutamine residues (acyl modifies the anti-deformation ability of the oat dough.
donor) and a variety of primary residues (acyl- Similarly, rye dough also shows a stronger elastic char-
acceptors) (Gallagher, 2009; Jaros et al., 2005). acter than a viscous one due to protein aggregation
Formation of an isopeptide bond between a free (probably due to isopeptide cross-links) (Beck et al.,
amine group (e.g. protein- or peptide-bound lysine resi- 2011). The relative viscosity significantly decreases by
dues) and the g-carboxamide group of protein- or pep- 40% with an increase in TG concentration (Beck et al.,
tide-bound glutamine residues causes the formation of 2011). At the same time, the relative elasticity of rye
high molecular weight polymers. It seems to be the pre- dough significantly increases by 31%. These properties
dominant reaction caused by TG in nature (Jong and almost remain constant when TG concentration is
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Table 2. Effect of TG on the functional properties of wheat and gluten-free dough
Characteristics Cereal Effect Authors

0
Pasting Buckwheat Increased elastic (G ) and viscous Kohajdova and
modulus and viscoelasticity Karovicova (2009)
Rice Increased elastic (G0 ) and viscous Renzetti et al. (2008a)
modulus
Rice Only a significant increase of the Marco and Rosell (2008a)
elastic modulus
Oat Increased elastic (G0 ) and viscous Huang et al. (2010)
modulus; elastic modulus is
always higher than the viscous
ones
Rye Stronger character of elastic Beck et al. (2011)
modulus than the viscous ones;
increased relative elasticity by
31% and relative viscosity
Pre-gelatinized Stronger character of elastic Onyango et al. (2010)
cassava þ sorghum modulus than the viscous ones;
increased relative elasticity and
relative viscosity
Thermal Oat Slight variation in transition onset Huang et al. (2010)
and peak temperatures;
improvement of dough stability
and increased enthalpy
Wheat þ barley þ soy Decreased enthalpy Ahn et al. (2005)
Mixing Oat Decreased water absorption and Huang et al. (2010)
increased torque peak, develop-
ment time and stability
Wheat Decreased water absorption Basman et al. (2002)
Buckwheat Increased torque peak, develop- Han et al. (2013)
ment time and stability and
decreased water absorption
Oat Structural changes of oat globulin, Siu et al. (2002)
no significant differences in oat
starch and cooking stability
Quantification of Oat Decreased number of free amino Huang et al. (2010)
free amino acid acid groups
groups Rice Decreased number of free amino Bonet et al. (2005) and
acid groups Gujral and
Rosell (2004)
Buckwheat Decreased number of free amino Han et al. (2013)
acid groups
Sodium Dodecyl Oat Globulin and avenin: good sub- Huang et al. (2010)
Sulphate-PolyAcrylamide Gel strates for TG. Cross-linking of
Electrophoresis (SDS-PAGE) avenalin and glutelin by TG
above 1000 U/kg for different reasons, namely (Beck Beck et al. (2011) showed that zero shear viscosity
et al., 2011): (a) no additional protein aggregation in was observed for the controls in rye dough production
the protein network of the dough system occurs, (b) due to the relatively low molecular weight of proteins
additional protein network formation occurs, but because of the lack of further cross-links by isopeptide
these changes do not contribute to significant changes bonds or rather no further protein aggregation. Due to
in dough rheology any more or (c) other enzymatic the above reasons, the relative viscosity of gluten-free
reactions (deamination of glutamine to glutamic acid) dough steadily rises with increasing TG until 1000 U/kg
negate additional protein aggregation. (Beck et al., 2011). These results are consistent with the
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work reported by Onyango et al. (2010) on pre-gelati- dough extensibility increases due to modification of
nized cassava and sorghum. Marco and Rosell (2008a) the cross-link between the oat proteins by TG, thereby
reported that there is only a significant increase on the increasing the stability of the protein network. The rise
elastic modulus of the rice–protein blends. Other par- in water-holding capacity is attributed to the cross-
ameters such as viscous modulus and viscoelasticity do linking after TG addition that changes in secondary
not show significant difference. Therefore, there is an structure or, possibly, due to changes in protein hydro-
increase in G0 (elastic modulus) values when cereal pro- phobicity from the formation of glutamic acid residues
teins are treated with TG (Demirkesen et al., 2010; from glutamine hydrolysis (Gerrard et al., 1998).
Larre et al., 2000). Nevertheless, TG has negative Structural changes in TG-treated oat globulin were
effects on corn flour where its application is detrimental also reported by Siu et al. (2002). No significant differ-
for the elastic properties of the dough (Arendt et al., ence has been noted in the oat starch after TG treat-
2008). ment (Siu et al., 2002). TG does not significantly affect
The thermal properties of oat dough measured by the setback value and cooking stability of the oat flour
differential scanning calorimeter show that a single (Siu et al., 2002). There is only an increase in cooking
endothermic peak is obtained between 60 and 70 C stability at a TG level of 1.5%. The presence of TG
after TG addition (Huang et al., 2010). A slight vari- promotes an increased torque peak and decreases the
ation in transition onset temperature (To) and transi- water absorption of oat dough (Huang et al., 2010).
tion peak temperature (Tp) is observed between samples These findings are mainly attributed to the cross-links
treated with TG compared to a control sample that catalysed by TG.
does not contain TG (Huang et al., 2010). This vari- The reaction between an "-amino group on protein
ation indicates that TG improves the thermal stability bound lysine residues and a g-carboxamide group on
of the dough (4.53–5.03 min). Furthermore, enthalpy protein bound glutamine residues leading to covalent
(H) of the dough samples significantly increases cross-linking of the proteins is catalysed by TG (Huang
(0.56–0.70 J/g) with TG treatment (Huang et al., et al., 2010). The implication of the amino groups in the
2010) because flour is heterogeneous material, hence cross-linking reaction reduces the number of these
the value of enthalpy reflects a combination of the tran- groups. This decrease in the number of the free amino
sition of all components in the flour sample. These find- groups is noticed when rice (Demirkesen et al., 2010)
ings are not consistent with those of Ahn et al. (2005) and buckwheat (Han et al., 2013) proteins are treated
who reported that TG’s impacts on protein denatur- with TG. However, no significant change of the number
ation of pure protein samples leading to a decrease in of free amino groups is observed when TG level exceeds
enthalpy due to protein unfolding. Therefore, the lower 1.0% (Han et al., 2013). This is relatively due to the
H values of soy flours or wheat–soy blends are due to limited reaction of glutamine coming from the add-
the high protein content of soy (Huang et al., 2010). itional TG in the dough. The free amino groups of
Larre et al. (2000) similarly demonstrated that TG has the proteins of oat flour are measured to assess the
significant impacts on the thermal stability of gluten. effect of TG. The protein modification made due to
This is mainly due to the covalent cross-linkage pro- TG addition is measured by changes in the number of
moted by the network and enzyme, which renders them free amino groups before and after TG treatment
insensitive to the temperature (Huang et al., 2010). (Huang et al., 2010). A reduction in the number of
The mixing analysis of oat dough by Mixolab high- free amino groups is progressively noticed when TG
lights that the water absorption decreases (66.1–65.2%) is added up to 1.0%. No significant differences in the
as the level of TG increases (Huang et al., 2010). number of free amino groups are observed beyond
Similar findings are reported by Basman et al. (2002) 1.0% (Huang et al., 2010). Gujral and Rosell (2004)
and Han et al. (2013) on wheat and buckwheat dough. reported similar findings and proposed that this phe-
These results are attributed to acyl-transfer reactions nomenon is due to a low amount of lysine limiting the
that introduce new functional groups leading to action of the additional TG. Although oat protein is
changes in the structure, charge and hydrophobicity rich in lysine at a TG level of 1.5%, the number of free
of the buckwheat proteins (Han et al., 2013). The amino groups decreased significantly because oat sam-
increase in developing time (0.65–0.82 min) and stabil- ples have a low protein content, which limits the TG
ity (4.53–5.03 min) indicates that the elasticity of the oat action (Huang et al., 2010).
dough is increased by the TG action (Huang et al., By promoting the cross-linking effect on different
2010). Similar results are reported by Han et al. flours (Autio et al., 2005; Bauer et al., 2003; Gerrard
(2013) on buckwheat dough. These findings are due et al., 2001; Larre et al., 2000; Rosell et al., 2003), TG
to the presence of storage proteins (2S albumin and widely modifies bread rheological properties (Caballero
8S and 13S globulin) of buckwheat flour that are et al., 2007) as summarized in Table 3. According to
cross-linked after a TG treatment. Moreover, the Caballero et al. (2007), TG significantly decreases loaf
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Table 3. Effect of TG on the functional properties of wheat gluten-free breads
Cereal Effect Authors
Wheat Decreased loaf specific volume, but no Caballero et al. (2007)

change in the loaf shape; increased rate of
bread staling during storage, limitation
of availability for starch and acceleration
of retrogradation, increased crumb
hardness, cohesiveness, gumminess,
chewiness and resilience
Extension of bread shelf-life and sensory Collar and Bollain (2005)
deterioration
Rice Improvement of volume, decreased hard- Shin et al. (2010)
ness, but no effect on springiness
Pre-gelatinized cassava Increased crumb firmness, no effects on Onyango et al. (2010)
þ sorghum springiness, cohesiveness and resilience
Oat, sorghum and tef No effect Arendt et al. (2008) and
Renzetti et al. (2008a)
Oat Increased crumb hardness, cohesiveness, Salmenkallio-Martilla et al. (2004)
gumminess, chewiness and resilience
specific volume but results in no change in the loaf their allergenicity (Zhu and Tramper, 2008). The single
shape. TG addition in rice flour strongly improves presence of TG increases the rate of bread staling
volume (565–633 ml) (Shin et al., 2010). The highest during storage (Caballero et al., 2007) and specifically
specific volume of rice bread is obtained with TG at affecting bread hardness, chewiness and gumminess. It
1 U/g (Autio et al., 2005). is due to the interaction between starch granules and
The incorporation of TG leads to a significant the protein network actively contributing to crumb
increase in crumb hardness, cohesiveness, gumminess, firming. The microscopic analysis of bread crumb
chewiness and resilience of wheat bread (Caballero points out significant differences in the starch–protein
et al., 2007). Similar results have been reported matrix during the course of storage (Blaszczak et al.,
by Salmenkallio-Marttila et al. (2004) on oat bread. 2004). TG promotes the affinity for water and therefore
In contrast, the hardness of rice bread decreases limiting water availability for starch and accelerating its
(6.836–5.731 N) with the addition of TG but the retrogradation (Caballero et al., 2007). Hence, this
springiness is not affected (Shin et al., 2010). starch retrogradation induced by TG decreases bread
Increasing TG concentration significantly increases shelf-life.
chewiness (9.190–12.133 N) and crumb firmness
(17.696–21.808 N) but does not affect springiness,
Effect of CG on the functional properties of
cohesiveness and resilience of pre-gelatinized cassava
gluten-free dough and bread
and sorghum bread (Onyango et al., 2010). These vari-
ations may be due to the molecular weight of the pro- Cyclodextrinase or CG (EC 2.4.1.19) acts at a pH range
teins formed during the cross-linking action of of 5.0–5.5, the temperature should not exceed 80–90 C
this enzyme (Marco et al., 2007, 2008). Although TG and the enzyme reaction can be terminated by lowering
is shown among other enzymes to enhance gluten- the pH. It catalyses four different reactions: cyclization,
free bread texture depending on the raw material, coupling, disproportionation and hydrolysis (Ohnishi
Arendt et al. (2008) reported that no impact of TG et al., 1997; Feng et al., 2011) with the production of
could be observed on breads from oat, sorghum or cyclodextrins at the end of these reactions (Figure 1).
tef. Similar results have been reported by Renzetti Cyclodextrins are made from the hydrolysis and cyc-
et al. (2008b). lization of starch, releasing closed circular molecules of
TG extends bread shelf-life by lowering crumb stal- six, seven or eight glucose units, referred to as a-, b- or
ing kinetics and sensory deterioration during storage g- cyclodextrin, respectively (Gujral et al., 2003). These
when used in combination with a-amylase (Collar and molecules possess a polar surface responsible for the
Bollain, 2005). However, a certain study revealed that aqueous solubility and a hydrophobic inner core
TG increases the shelf-life of certain foods and reduces (Gujral et al., 2003). Thus, their most characteristic
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Figure 1. Proposed model of the events taking place in the CG catalysed reactions. (a) Disproportionation, (b) coupling,
(c) cyclization. The different CG domains are indicated (A, B, C, D and E). 1 and 2 indicate the maltose binding sites on the
E-domain. The triangle indicates the cleavage site in the active site. Circles represent glucose residues; acceptors resi-
dues are represented in black. (Adapted from Feng et al. (2011)).
property is that they have a hydrophilic exterior. This behaviours of dough (Gujral et al., 2003). Table 4 sum-
property allows them to dissolve in water while having marizes the effect of CG on the functional properties of
a hydrophobic cavity that forms inclusion complexes rice dough.
with a wide variety of hydrophobic guest molecules. The incorporation of CG lowers dough consistency
The cyclodextrins form complexes with fatty acids when increasing CG to 40 ml/100 g flour. This indicates
and emulsifiers influencing the rheological properties that CG brings about some breakdown in the starch
of starch and the functionality of the resultant starch during the mixing process (Gujral et al., 2003). The
(Rosell, 2001). Consequently, the pasted starch con- elastic modulus of rice dough is higher than the viscous
taining cyclodextrins lower elastic and viscous modulus (Gujral et al., 2003). The CG addition to the
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Table 4. Effect of CG on the functional properties of rice amylose complex as suggested by Gujral et al. (2003)
dough and bread prepared using rice flour (Gujral et al., and Liang et al. (2002). From the effect of the highest
2003) concentration of CG on the gelatinization property of
Product Effect rice flour, it is predicted that a softer crumb texture is
obtained by adding this enzyme, which agrees with
Dough Decreased consistency, elastic modulus, breadmaking results (Gujral et al., 2003). The cyclizing
peak viscosity and final viscosity activity of the CG promotes an additional impact on the
Increased viscoelasticity and breakdown gelatinization behaviour of rice flour. It is mainly due to
Elastic modulus higher than the viscous the fact that the resulting cyclodextrins form complexes
modulus
with different compounds.
Bread Increased specific volume by 73%, shape According to Rosell (2001), CG degrades starch of
index and volume
gluten-free dough by its hydrolysing and cycling activ-
Negative correlation between crumb
firmness and specific volume
ities. The hydrolysis reaction releases cyclodextrins
Decreased crumb firmness by 53%, which are able to form complexes with lipids and pro-
resulting in soft bread teins. The necessary substrates for the complex forma-
Extension of shelf-life by retarding tion between lipids and proteins with cyclodextrins are
amylopectin retrogradation provided by the cyclization reaction (Rosell, 2001).
Therefore, the hydrophobic environment of gluten-
free dough is reduced by CG through starch hydrolys-
ing and cyclizing activities and also through the
dough lowers the elastic modulus and complex viscosity hydrolysis products that can form complexes with a
of the dough but does not seem to influence the viscous variety of solid, liquid and gaseous compounds.
modulus. According to Gujral et al. (2003), the tan Preliminary experiments indicated that CG has posi-
(G00 /G0 ) or viscoelasticity increases in the presence of tive impacts on rice bread volume (Gujral et al., 2003).
the enzyme, suggesting that the relative contribution Increasing the CG concentration from 0 to 20 ml/100 g
of the solid character (G0 ) decreases. It acts on the of flour increases the specific volume by 73% (Gujral
damaged starch during the mixing process and proofing et al., 2003). This effect is probably due to the release of
time (30 C). This action brings about some hydrolysis, fermentable sugars utilized by the yeast, as a result of
which impacts the dough rheology. A decrease in the the hydrolysis of starch, which is catalysed by CG.
elastic modulus and an increase in the tan in wheat Regarding the crumb texture, the crumb firmness
flour dough from sprouted wheat flours are highlighted shows a negative correlation with specific volume.
by Singh et al. (2001) and are due to higher amylase and The incorporation of increasing CG dosage consider-
protease activities. ably lowers firmness by 53%, obtaining a very soft
CG incorporation (20 ml/100 g of rice flour) lowers bread crumb (Gujral et al., 2003). The identical
the peak viscosity (2424 cP) and slightly affects the decrease in the crumb firmness is obtained with the
final viscosity (3147 cP), indicating that the enzyme addition of a-amylase, but the crumbs are very sticky
acts on the starch, hence lowering the viscosity (Gujral et al., 2003). Hence, the improvement produced
(Gujral et al., 2003). According to Gujral et al. (2003), by CG can be attributed to the starch hydrolysis that
CG also increases the breakdown (1250 cP), indicating yields fermentable sugar but also to the cyclization of
that the paste is less resistant to heating and shear stress the hydrolysis products, which form complexes with
because starch is hydrolysed. When adding CG at lipids and also proteins (Gujral et al., 2003). CG pos-
higher levels (40 ml/100 g of rice flour), a further sesses multiple catalysing activities; therefore, the
decrease of the peak viscosity (2136 cP) along with an improvement of rice bread results from the combined
increase of the breakdown (1399 cP) is observed. The effect of those activities. Despite CG having a cyclizing
setback defined as the difference between the peak vis- activity, it is important to validate that the breadmak-
cosity and the viscosity at 50 C is related to the starch ing conditions are favourable for that reaction to take
retrogradation. It relates to amylose helix interaction place. No detectable amount of cyclodextrins is
and is one of the most important parameters in predict- observed in the crumb from rice bread obtained in
ing rice bread characteristics (Nishita and Bean, 1979). the absence of CG. The cyclodextrin dosage increases
CG decreases the final viscosity, although the setback by increasing the CG concentration. The presence of
reduces at 40 ml/100 g of flour (Gujral et al., 2003). It is cyclodextrins validates the cyclizing activity of CG
possible that at a low enzyme dosage, some hydrolysis during the breadmaking process. Therefore, the hydro-
products are associated during cooling as the amylose lysing activity of the CG during breadmaking is identi-
does, and at high enzyme level a high amount of cyclo- cal to the effect of a-amylase. The starch hydrolysis
dextrins is present, which physically interferes with the yields fermentable sugars which are metabolized by
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Table 5. Effect of CMC on the functional properties of gluten-free products
Product Cereal Effect Authors
Dough Rice Increased water absorption Leon et al. (2002)

Increased development time, elastic modulus, Lazaridou et al. (2007)
viscous modulus and viscoelasticity
Elastic modulus higher than viscous modulus
Increased development time Sivaramakrishnan et al. (2004)
Decreased water absorption Bertolini et al. (2005)
Increased consistency Sciarini et al. (2010)
Bread Rice Increased volume Lazaridou et al. (2007)
No significant effects on crumb firmness and
crust yellowness
Decreased hardening rate, retarding
staling Increased ‘a value’ or
redness colour of crust and crumb
Lighter colour of crust Sciarini et al. (2010);
Mezaize et al. (2009)
Fonio (acha) Increased loaf volume, crumb texture, crust Jideani et al. (2007)
colour, crumb colour, general acceptability
No significant effect on
specific volume
Gluten free Decrease of crumb hardness Arendt et al. (2008)
Gluten free Comparable to wheat bread in terms of sensory Ylimaki et al. (1991)
attributes
yeast. Triglyceride molecules and lipids with inclusion

Effect of CMC on the functional properties
complexes reducing interfacial tension are formed by
of gluten-free dough and bread
cyclodextrin molecules which act as emulsifiers (Liang
et al., 2002; Shimada et al., 1992). The softening effect CMC (E 466) is a derivative of cellulose with carbox-
of the emulsifiers on the crumb of wheat bread is well ymethyl groups bound to some of the hydroxyl groups
known (Collar et al., 1998). Furthermore, cyclodextrins present in the glucopyranose monomers that form the
possess the capability to interact with hydrophobic pro- cellulose backbone. Its molecular structure is based on
teins, leading to increased solubility (Lee and Fennema, the b-(1!4)-D-glucopyranose polymer of cellulose.
1991). The complexes with the hydrophobic proteins Different formulations could exist with different
(globulin and glutelin) of rice formed by cyclodextrins degrees of substitution, but it is mainly in the range
increase their solubility (Gujral et al., 2003) improving 0.6–0.95 derivatives per monomer unit (Lsbu, 2013).
CO2 retention, increasing volume and enhancing tex- The functional effects of CMC on dough are high-
ture of bread. Table 4 summarizes the effect of CG lighted in Table 5. The addition of hydrocolloids such
on the functional properties of rice bread. as CMC increases the water absorption of rice dough.
CG contributes to extending the shelf-life of rice It is attributed to the hydrophilic character of these
bread. Its shelf-life extension is due to its ability to polymers (Leon et al., 2002). The highest absorption
decrease amylopectin retrogradation during storage is observed for CMC (63.4%), followed by the control
through its hydrolysing and cyclizing activities (Gujral at 60.5%. The DDT increases with CMC incorporation
et al., 2003). This anti-staling effect is attributed to the (26.5%) whereas control decreases it (4.0%). Thus,
low molecular weight dextrins produced as a result of CMC exhibits a stronger negative effect on the farino-
starch hydrolysis. Those dextrins interfere with the cap- graph curve (with an increase of DDT, 26.5 min) com-
ability of the amylopectin to retrograde (Defloor and pared to the control (4.0 min) (Lazaridou et al., 2007).
Delcour, 1999; Leon et al., 2002; Lin and Lineback, These findings are consistent with the studies performed
1990; Rojas et al., 2001), or with other interactions by Sivaramakrishnan et al. (2004) in rice flour fortified
also, namely starch–protein or protein–protein with the addition of 4.5% HPMC. However, Bertolini
entanglement involved in firming (Lin and Lineback, et al. (2005) showed that a non-starch polysaccharide
1990; Martin and Hoseney, 1991). such as CMC decreases water absorption of rice starch
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gel. This addition of non-starch polysaccharides such as The addition of CMC does not significantly affect
CMC to starch–water systems limited the hydration of the crumb firmness of rice bread compared to control
the starch and, as water had a plasticizing effect in formulations (Lazaridou et al., 2007). The strengthen-
amorphous regions of the starch, the mobility of the ing effect of CMC on crumb structure appears to be
plasticizer was also restricted. Therefore, non-starch consistent with the low rigidity highlighted by dough
polysaccharides may have an ‘anti-plasticizing’ effect containing it. Thus, a low rate of crumb hardness is
(Bertolini et al., 2005). observed for breads supplemented by CMC.
The hydrocolloid incorporation increases the The sensory evaluation by an untrained consumer
dynamic elastic modulus (Lazaridou et al., 2007). panel revealed that gluten-free bread containing 2%
CMC renders rice dough more elastic (70 BU) than CMC is highly acceptable because of its low crumb
the dough control (60 BU). However, the increasing hardening compared to the ones containing other
effect of hydrocolloid level on G0 values is not clear hydrocolloids (Arendt et al., 2008). In addition, the
because the added water also increases and CMC loaf of acha bread with 4% CMC is significantly
becomes stronger (higher G0 values) with increasing better in terms of appearance, crust colour, crumb tex-
concentration affecting the rheological properties of ture, crumb colour and general acceptability compared
dough more than the increasing content of water to other loaves of acha bread (Jideani et al., 2007). The
(Lazaridou et al., 2007). Although gluten-free dough crumb texture, crumb colour and general acceptability
exhibits an elastic modulus higher than the viscous of acha bread containing 4% CMC is not significantly
modulus, the dough made with CMC has a higher vis- different from wheat bread (Jideani et al., 2007). In
cosity than the ones containing no hydrocolloid addition, Ylimaki et al. (1991) found that gluten-free
(Lazaridou et al., 2007). Moreover, the viscoelasticity bread containing CMC is comparable to a reference
of gluten-free dough is lower with CMC. The consist- wheat bread on sensorial attributes from a trained
ency of gluten-free dough supplemented by CMC panel.
is greater (419.5) than the control (285.4) The colour analysis of the crust of gluten-free bread
(Sivaramakrishnan et al., 2004). reveals that bread supplemented with CMC has a
Table 5 summarizes the effect of CMC on the func- lighter crust compared to the control (Sciarini et al.,
tional properties of gluten-free bread. Thus, hydrocol- 2010). This could be attributed to the effect of the
loids such as CMC improve the volume of gluten-free hydrocolloid on water distribution which affects the
formulations. The greatest volume is exhibited for rice Maillard reaction and caramelization. Similar findings
bread supplemented with CMC (267 cm3/100 g bread) were obtained by Mezaize et al. (2009) studying the
among hydrocolloids used (Lazaridou et al., 2007). colour of gluten-free breads. The colour analysis of
This improvement is attributed to the increase of rice bread demonstrated that the redness value of
dough viscosity by hydrocolloids leading to the crust is higher for a CMC formulation as compared
enhanced dough development and gas retention to that of the control (Lazaridou et al., 2007). No sig-
(Rosell et al., 2001). The modified polysaccharide nificant difference in crust yellowness was found. The
derivatives such as CMC contain hydrophobic groups presence of CMC at 2% concentration showed a sig-
imparting additional properties which increase inter- nificant difference of the redness parameter for crumb
facial activity of the dough system during proofing among gluten-free breads (Lazaridou et al., 2007).
and producing gel networks on heating during the The analysis of stored bread quality showed that a
breadmaking process. These network structures low rate of hardening is observed for bread supple-
increase viscosity and strengthen the boundaries of mented with CMC when added at a concentration of
the expanding cells in the dough, thereby increasing 2%. After 3 days of storage, the softest crumb is asso-
gas retention during baking which leads to a better ciated with CMC-supplemented bread (Lazaridou
loaf volume (Bell, 1990). Furthermore, the addition of et al., 2007).
CMC on Fonio or Acha flour increases the loaf volume
by 40–59.5% of the resulting bread but its specific loaf
Effect of HPMC on the functional properties
volume does not significantly differ (2.60–2.73 ml/g)
of gluten-free dough and bread
(Jideani et al., 2007). Small bread volume of control
(219 cm3/100 g bread) might be attributed to highest HPMC (E 464) is produced by the addition of methyl
strength and elasticity of dough which cause a limited and hydroxypropyl groups to the cellulose chain
and slow expansion of the gas cells during proofing (Shhuiguang, 2013) leading to a polymer with high sur-
similar to bread supplemented with xanthan gum face activity and unique properties concerning its
(Lazaridou et al., 2007). Consequently, the dough hydration–dehydration characteristics in the solution
becomes too rigid to incorporate gases (Lazaridou state and during temperature changes (Kohajdova
et al., 2007). and Karovicova, 2009). Its effects on the functional
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Table 6. Effect of HPMC on the functional properties of wheat and gluten-free products
Product Cereal Effect Authors
Dough Gluten free Improvement of water absorption Huttner and Arendt (2010)
and gas retention
Wheat Increased viscosity Barcenas and Rosell (2005)
Bread Gluten free Increased crumb moisture content Bell (1990) and Dziezak (1991)
Improvements of sensorial properties, Kohajdova and karovicova (2009)
crumb texture and softness
Wheat Increased volume Rosell et al. (2001)
Decreased crumb hardening rate and staling Collar et al. (2001) and Guarda et al. (2004)
Improvement of crumb texture (soft crumb) Barcenas and Rosell (2005)
properties of wheat and gluten-free dough are sum- crumb texture and softness without any adverse
marized in Table 6. The incorporation of HPMC impact on the palatability of the fresh product
improves gas retention and water absorption of (Kohajdova and Karovicova, 2009).
gluten-free dough, a property which is usually con- The microstructure analysis showed a possible inter-
ferred by gluten (Huttner and Arendt, 2010). It also action between HPMC and the bread constituents
increases the viscosity of aqueous systems interfering (Barcenas and Rosell, 2005) and its capacity to interact
with the diffusion phenomena (Barcenas and Rosell, with the effective water present in the system (Schiraldi
2005). According to Bell (1990), the substitution of et al., 1996) thereby performing an anti-staling func-
the hydroxyl groups of cellulose by methoxyl and tion. The softening impact of HPMC is attributed to
hydroxypropyl increases the water solubility and the its water retention ability and its inhibition of amylo-
affinity to the non-polar phase enhancing the hydro- pectin retrogradation since HPMC preferentially binds
philic character of HPMC. This double role allows to starch (Collar et al., 2001), avoiding starch–gluten
keeping the dough uniform and the emulsion stable interactions and therefore reducing crumb hardening
during breadmaking. Therefore, it increases the water rate and staling.
absorption of dough. The improvement of gas retention The improving effect of HPMC on the sensory qual-
is attributed to the formation of interfacial films at the ity of bread is attributed to its impact on the crumb
boundaries of gas cells conferring some stability against texture through the production of softer crumbs
gas expansion (Huttner and Arendt, 2010). The optimal (Barcenas and Rosell, 2005).
level of HPMC in gluten-free bread formulation con-
sisting of rice flour and potato starch was determined to
CONCLUSION
be 2.2% (McCarthy et al., 2005). The addition of
HPMC in gluten-free bread formulations increases the The cornerstone treatment for patients with coeliac’s
moisture content of the crumb (Bell, 1990; Dziezak, disease is a lifelong elimination of gluten in their diet.
1991), bread volume (Rosell et al., 2001), reduces the Hence, the biggest challenge for food technologists
crumb hardening rate (Guarda et al., 2004) and remains the production of high quality gluten-free
improves crumb texture (Barcenas and Rosell, 2005) bread with properties similar to that of
and sensorial properties (Kohajdova and Karovicova, gluten-containing bread. The majority of breads that
2009) as summarized in Table 6. The capability of are currently on market shelves exhibit a very poor qual-
HPMC to improve bread volume/mass ratio is due to ity due to the lack of gluten. Good quality gluten-free
the release of water molecules allowing a stronger inter- bread can only be made with polymeric substances
action between the chains when exposed to higher tem- which mimic the viscoelastic behaviours of gluten.
peratures. As a result, there is the creation of a Dairy-based ingredients are shown to be the most pro-
temporary network that disintegrates under cooling mising among protein-based ingredients in the improve-
conditions (Bell, 1990). Dough expands during ment of gluten-free bread properties. Water is one of the
baking, the gas losses are reduced and volume increased most important ingredients in any gluten-free formula-
due to gas cells strengthening of dough created by the tion and therefore needs to be optimized to achieve opti-
HPMC network. This barrier for gas diffusion of mal results. Recently, research has focused on the
HPMC decreases water vapour losses (Bell, 1990) and application of TG, CG, CMC and HPMC to improve
increases the final moisture content of the loaf the texture of gluten-free bread. TG in gluten-free bread
(Barcenas and Rosell, 2005) and provides better products modifies the viscoelasticity properties of the
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dough and improves the rheological behaviours and gluten proteins of wheat I: Biochemical analysis. Cereal
shelf-life of the resulting bread. CG produces a reduc- Chemistry 80: 781–786.
tion in dough consistency and elastic modulus and Beck M, Jeckle M, Selmair PL, Koehler P and Becker T.
enhances specific volume, shape index, crumb texture (2011). Rheological properties and baking performance
and shelf-life of bread. CMC increases dough elasticity, of rye dough as affected by transglutaminase. Journal of
Cereal Science 54: 29–36.
bread volume and does not alter the crumb firmness.
Bell DA. (1990). Methycellulose as a structure enhancer in
HPMC is an important ingredient for gluten-free
bread baking. Cereal Food World 35: 1001–1006.
bread production because of its ability to mimic the Bernadin JE and Kasarda DD. (1973). The microstructure of
viscoelasticity properties of gluten to a certain extent. wheat protein fibrils. Cereal Chemistry 50: 529.
It reduces staling, improves water binding and overall Bertolini AC, Creamer LK, Eppink M and Boland M. (2005).
structure of the resulting bread but further research is Some rheological properties of sodium caseinate-starch
required to optimize the application of this hydrocolloid gels. Journal of Agricultural and Food Chemistry 53:
in gluten-free systems. Although research on gluten-free 2248–2254.
bread is still in its infancy, researchers should develop Blanshard JMV. (1986). The Significance of the Structure and
the high-quality products by the right selection and com- Function of the Starch Granule in Baked Products. London:
bination of the additives in order to obtain breads with a The Royal Society of Chemistry.
desirable quality. Blaszczak W, Sadownska J, Rosell CM and Formal J. (2004).
Structural changes in the wheat dough and bread with the
addition of alpha-amylases. European Food Research and
ACKNOWLEDGEMENTS
Technology 219: 348–354.
The authors wish to acknowledge the Cape Peninsula Bonnet A, Caballero PA, Gomez M and Rosell CM. (2005).
University Research Fund and the National Research Microbial transglutaminase as a tool to restore the
Foundation for funding this work. functionality of gluten from insect-damaged wheat.
Cereal Chemistry 82(4): 425–430.
FUNDING Brown J. (1993). Advances in Breadamaking Technology.
This research received no specific grant from any New York, NY: Blackie Academic and Professional.
funding agency in the public, commercial, or not-for-profit Caballero PA, Gomez M and Rosell CM. (2007).
sectors. Improvement of dough rheology, bread quality and
bread shelf-life by enzymes combination. Journal of Food
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Advances in gluten-free bread technology

PH Nitcheu Ngemakwe1, M Le Roes-Hill2 and VA Jideani1

gluten is also important for crumb structure of wheat

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Table 1. Gluten-free bread production processes

Cereal flour Unit operation Authors

Nitcheu Ngemakwe et al.

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Table 2. Effect of TG on the functional properties of wheat and gluten-free dough

Characteristics Cereal Effect Authors

Nitcheu Ngemakwe et al.

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Table 3. Effect of TG on the functional properties of wheat gluten-free breads

Cereal Effect Authors

Wheat Decreased loaf specific volume, but no Caballero et al. (2007)

Nitcheu Ngemakwe et al.

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Nitcheu Ngemakwe et al.

Table 5. Effect of CMC on the functional properties of gluten-free products

Product Cereal Effect Authors

Dough Rice Increased water absorption Leon et al. (2002)

yeast. Triglyceride molecules and lipids with inclusion

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Product Cereal Effect Authors

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