Accepted Manuscript

Title: Development of gluten-free bread using Guar gum and

Transglutaminase

Author: Mehrdad Mohammadi Mohammad-Hossain Azizi

Tirang R. Neyestani Hedayat Hosseini Amir Mohammad
Mortazavian

PII: S1226-086X(14)00311-6
DOI: http://dx.doi.org/doi:10.1016/j.jiec.2014.06.013
Reference: JIEC 2089

To appear in:

Received date: 1-2-2014

Revised date: 31-5-2014
Accepted date: 9-6-2014

Please cite this article as: Mehrdad Mohammadi, Mohammad-Hossain Azizi, Tirang
R.Neyestani, Hedayat Hosseini, Amir Mohammad Mortazavian, Development of
gluten-free bread using Guar gum and Transglutaminase, Journal of Industrial and
Engineering Chemistry http://dx.doi.org/10.1016/j.jiec.2014.06.013

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 Development of gluten-free bread using Guar

gum and Transglutaminase

Mehrdad Mohammadia, Mohammad-Hossain Azizib, Tirang R.

Neyestanic‫٭‬, Hedayat Hosseinid, Amir Mohammad Mortazaviand

t
ip
cr
a
Department of Food Technology Research, National Nutrition and Food

Technology Research Institute, Faculty of Nutrition Sciences and Food

us
Technology, Shahid Beheshti University of Medical Sciences, Tehran,

Iran. an
b
Department of Food Science and Technology, Faculty of Agriculture,
M
Tarbiat Modarres University, Tehran, Iran.
c
Department of Nutrition Research, National Nutrition and Food
d

Technology Research Institute, Faculty of Nutrition Sciences and Food

e

Technology, Shahid Beheshti University of Medical Sciences, Tehran,

pt

Iran.
d
Department of Food Science and Technology, National Nutrition and
ce

Food Technology Research Institute, Faculty of Nutrition Sciences and

Food Technology, Shahid Beheshti University of Medical Sciences,

Ac

Tehran, Iran.

‫٭‬
Correspondence to: Postal Code: 1981619573, P.O. Box: 19395-

4741.

1
Page 1 of 23
 Abstract

The effects of guar gum (20 and 30 g kg-1) in combination with

microbial transglutaminase (TGase) (0, 1 and 10 u/g pro) on the quality

parameters of gluten-free bread, based on rice flour were investigated.

Incorporation of guar gum significantly increased specific volume,

t
leading to lower crumb hardness on the baking day compared with

ip
control (P<0.05). Addition of 1 u/g pro of TGase caused appropriate

cr
crumb texture and increase in Tgase concentration yielded higher

us
crumb hardness. G3T1 formula, containing 30 g kg-1 of guar gum and 1

u/g pro of TGase was the best formulae, compared to all the others.
an
Keywords: gluten-free; bread; guar gum; microbial transglutaminase;
M
gluten replacer.
d

Introduction
e

Coeliac is a lifetime intestinal disorder that dominates due to gluten

pt

ingestion in sensitive persons. Coeliac is one of the most common

ce

genetic diseases throughout the world. Gluten intake by coeliac

patients induces inflammation and swelling in the small intestine

Ac

followed by partial absorption of essential nutrients such as iron,

calcium and fat soluble vitamins, and increase in the ratio of weight

loss, diarrhea, anemia, fatigue, flatulence and folate loss [1,2]. Coeliac

disease not predominates only in the developed countries, but also is

reported in developing countries, increasingly.

2
Page 2 of 23
 Serological experiments in the 1980s in the developing countries

showed that coeliac prevalence in the Middle East countries/region

such as Iran is almost as high as in the European nations, and its

prevalence is 3-5% in high risk regions [3]. Intake of diets free of

protamine group proteins or gluten-free diets is the only coeliac curing

t
method all the time [4]. Wheat (gliadins), barley (hordeins), rye

ip
(secalins) and, for some people, oats (avenin) should be eliminated in

cr
gluten-free diets [5]. There is a doubt about oat elimination from gluten-

us
free diets and grains like corn and rice could be ingested by coeliac

patients [6].
an
Gluten is the most essential protein in wheat flour products, which

plays a significant role in the texture and appearance of final products

M
such as bread [7]. It has also a great role in bread-making process

because of its share in visco-elasticity, mixing resistance, dough

d

spread-ability, and gas holding capacity and good body structure in the
e

final product. For this reason, gluten substitution in bread is one of the
pt

most controversial challenges that grain technologists are facing with

ce

[8,9].

Simulation of visco-elastic properties of gluten in wheat dough is

Ac

done by using various flours and starches (e.g. rice, corn, cassava and

soy) and also gums, enzymes, soy proteins and egg-white. Cato et al.

studied the effect of carboxymethylcellulose (CMC), hydroxypropyl

methylcellulose (HPMC) and guar gums in gluten-free bread made by

rice flour and potato starch [10]. Curić et al. studied the effect of soy

flour on the protein fortification, mechanical properties, plasticity and

3
Page 3 of 23
 mixing resistance of gluten-free bread [11]. Mezaize et al. examined

the effect of guar gum and CMC instead of gluten in French bread and

reported that breads containing 20 g kg-1 guar gum have favorite

properties such as increased volume, softer crumb and favorable color

[12]. Anton and Artfield investigated the effect of different hydrocolloids

t
as binding agents and gluten alternative in bread made from corn

ip
starch [13].

cr
The common point between the above authors was that among

us
xanthan, carob, guar gum and tragacanth, xanthan showed the best

quality in bread making. Different methods including the use of

an
enzymes have been applied to propose the similar network to gluten

with regard to gas holding capacity and quality development of the final
M
product. For instance, Moore et al. studied the effect of different values

of TGase on protein network formation in gluten-free breads [14].

d

Bread is one of the most important foods in the daily diet of most
e

people throughout the world, and thus it needs to be investigated at

pt

any aspect. So this study focuses on the effect of microbial TGase and
ce

guar gum on the rheological properties of dough and the quality of

breads made by gluten-free rice flour, corn starch and soy flour.
Ac

Materials and methods

Raw materials

Corn starch (125 g kg-1 moisture, 30 g kg-1 protein) was obtained

from Farayand Co. (Karaj, Iran); rice flour (98 g kg-1 moisture, 84 g kg-1

protein), with the commercial name of ‘Tarom’, was purchased from a

4
Page 4 of 23
 local market (Rasht, Iran); defatted soy flour (80 g kg-1 moisture, 45 g

kg-1 protein) was obtained from Soya Sun Co. (Hashtgerd, Iran); inulin

was taken from Sensus Co. (Roosendaal, Netherlands); sodium

caseinate was purchased from Iran Caseinate Co. (Tehran, Iran);

vegetable oil was provided by Behshahr Industrial Co. (Tehran, Iran);

t
guar gum and TGase were obtained from Rama Industries (Deesa,

ip
India) and Activia Ajinomoto Co. (Hamburg, Germany) respectively;

cr
dried instant yeast was purchased from Khuzestan Yeast Co.

us
(Khuzestan, Iran) and DATEM (Diacetyl Tartaric Acid Ester of Mono-

and Di-glycerides of fatty acids) were obtained from Beldem

Co. (Bijgaarden, Belgium). an

Eighteen different gluten-free formulae were initially selected to
M
obtain the control gluten-free bread with most favorable rheological

and technological features and sensorial quality. Consequently, based

d

on the results obtained from the mentioned pilot study, all formulae in
e

this study included rice flour (200 gr), corn starch (150 gr), soy flour
pt

(50 gr), sodium caseinate (24 gr), oil (20 gr), sugar (20 gr), inulin (10
ce

gr), salt (7 gr), dried instant yeast (7 gr) and DATEM (1 gr). The control

was free from enzyme and gum. Guar gum was added at two levels
Ac

(20 g kg-1 and 30 g kg-1) and TGase was added at three levels (0, 1

and 10 u/g pro).

Bread preparation

In this work we prepared breads based on rice flour, corn starch and

soy flour, similar to “Pan bread”. Initially, the raw materials were

weighted and then the dry materials were poured into the Spiral Mixer

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Page 5 of 23
 M 80-Premium manufactured by Escher Mixers Co. (Schio, Italy) and

stirred for 5 min. Water was added so that all dough had the similar

consistency measured by farinograph (Brabender, Germany). Then oil

was added to the mixture and stirred for 7 min at 30 × g. Finally, the

obtained dough was mixed for 5 min at 90 rpm to make a homogenous

t
mixture. Then it was cut into pieces with 700 gr weights, and molding

ip
process was performed at greasy 9×19.5×10 cm dishes. Fermentation

cr
was carried out under 30 oC and 850 g kg-1 relative humidity for 45

us
min. Baking process was performed in Rototherm Oven - EN 6080 /

EN 80100 manufactured by Enkomak Bakery Machinery Co. (Anatolia,

an
Turkey) by 10 s steaming followed by heating at 230 oC for 30 min.

Finally, the breads were cooled at ambient temperature, sealed within

M
polypropylene coating and stored at 30 oC for 72 h. The values of

TGase and guar gum in gluten-free bread formulations are presented

d

in Table 1.
e

Bread quality evaluation

pt

The effects of guar gum and TGase on dough characteristics (water

ce

absorption of mixture, dough development time, dough resistance and

degree of softening after 10 min) were evaluated by farinograph and

Ac

using the method number AACC 54-21 [15].

Moisture content was measured by AACC 44-16 and crude protein

measurement was carried out by AACC 46-30 using correction index

of 6.25 for nitrogen-to-protein conversion [15]. Volume of the breads

was determined by the rapeseed displacement method of AACC 10-05

6
Page 6 of 23
 [15]. The obtained volume was divided to the bread's weight (gr) to

achieve the bread's specific volume (milliliter/gr).

Bread yield (%) was calculated by the following formula: (bread's

weight/flour-based mixture's weight)×100.

Dough yield (%) was measured by the amount of obtained dough

t
(gr) from 100 gr of raw materials. Crumb hardness was evaluated by

ip
instron (Model 088 MTCL, Rochdael, England) according to the

cr
method of AACC 74-09 [15]. The breads' staling was monitored after 0,

us
24, 48 and 72 h of storage using texture softness analysis. The more

compression was parallel to the higher energy consumed.

an
Sensory analysis was carried out by 5 trained assessors. The 2

slices of each formula were labeled with three-digit codes. The

M
assessors were asked to state their judgment of the formulae

individually in separate booths under white and yellow fluorescent

d

lights (similar to daylight). The order of presentation of the plates to

e

each assessor was different in a random sequence. For each

pt

characteristic an evaluation sheet was provided for each assessor.

ce

Bottled water at room temperature was also provided to clean the

palate between assessments of each of the samples. Scoring of each

Ac

characteristic was done as follows:

Form (0–10), crust color (0–15), crumb color (0–10), texture (0–15),

aroma (0–15), flavor and taste (0–20) and chewing ability (0–15).

This research was done in triplicate for each formula and all

measurements were repeated in duplicate.

Statistical analyses

7
Page 7 of 23
 The data were analyzed by one-way Analysis of Variance (ANOVA),

using SPSS 17 statistical package (SPSS Inc., Chicago, IL, USA). The

Duncan test was used to determine statistically significant differences

between the means. Data from the sensory evaluation were subjected

to the Kruskal-Wallis test. The Mann-Whitney U test was used to

t
determine the statistical significance between the means. P values

ip
<0.05 were considered as statistically significant for all comparisons

cr
[16].

us
Results and discussion

Farinograph characteristics
an
The results of farinograph are shown in Table 2. As shown, guar
M
gum increased the water absorption of flour mixture compared to the

control. The same result was observed by increasing of the gum

d

content from 20 to 30 g kg-1, but the difference was not significant at

e

95% confidence interval. Ćurić et al. observed the same results in guar
pt

gum incorporation [11].

ce

Addition of TGase increased significantly the water absorption of the

formulae compared to the control (P<0.05). Same result has been

Ac

reported by Marco and Rosell [17] and it was due to the role of TGase

as enzymatic cross-linker that induces protein network with high water

holding capacity [18].

The formulae containing 30 g kg-1 guar gum had the most dough

development time as well as the most water absorption. Guar gum

addition at the levels 20 g kg-1 and 30 g kg-1 lowered the degree of

8
Page 8 of 23
 softening compared to the control so that by increasing of guar gum

content from 20 to 30 g kg-1, the degree of softening decreased

insignificantly. Ćurić et al. observed the same results by addition of 30

g kg-1 guar gum to the mixture of rice flour and extruded corn starch

[11]. TGase addition and its level elevation from 1 to 10 u/g pro

t
decreased significantly the degree of softening compared to the control

ip
(P<0.05). Although one exception was observed in G2T1 formula and

cr
there was no significant difference. This exception was due to the

us
effect of Tgase on the formation of protein network same to gluten

network and polymeric protein formation due to free amino acid

an
binding resulted in texture maintenance and dough stability.

Moisture
M
As shown in Table 3, guar gum addition increased the moisture

content of breads compared to the control (P<0.05), due to high water

d

holding capacity of hydrocolloid [19]. Guar gum, classified as seed

e

mucilage, is a hydrocolloid from plant origin [20].

pt

Also addition of TGase and elevating its level increased the moisture
ce

content of breads in comparison to the control (P<0.05). Moore et al.

reported the similar results in this regard [14]. It is presumed that

Ac

cross-linking glutamine and lysine by TGase could lead to water

trapping and induce the increased water holding capacity [14]. On the

other hand, increased water absorption could be due to amide group

elimination from glutamine (resulted from TGase activity) and its

conversion into glutamic acid. Glutamic acid decreases hydrophobicity

and finally increases water absorption [21]. Some components such as

9
Page 9 of 23
 soy flour and dairy products could also increase water holding capacity

[22].

Dough yield

As shown in Table 3, guar gum and TGase increased significantly

the dough yield in comparison to the control (P<0.05). Ćurić et al.

t
reported the same results by addition of guar and xanthan gums to the

ip
mixture of rice and corn starch flours [11].

cr
Hydrocolloids are hydrophilic components. Water/hydrocolloid

us
interactions restrict water dispersion throughout the environment. Guar

gum is soluble in cool water and forms a viscous solution. So it

an
increases water absorption and dough yield.

Increasing of water holding capacity in gels by TGase activity [23]

M
and gluten-free systems [14] had been reported. It has been proposed

that amide group elimination from glutamine and cross-linking

d

glutamine and lysine could lead to the improvement of water holding

e

capacity [21,24].
pt

Bread yield
ce

According to Table 3, addition of guar gum at the levels of 20 g kg-1

and 30 g kg-1 increased the yield of bread in comparison to the control,

Ac

but the differences were not significant by the level elevation from 20

to 30 g kg-1. As stated earlier, hydrophilicity nature of gums maintains

bread's moisture after baking process. Increasing of the moisture

content of breads decreased weight loss and increased bread yield.

Ćurić et al. observed the same results by addition of guar and xanthan

gums to the formulations [11].

10
Page 10 of 23
 TGase addition at 1 and 10 u/g pro levels increased the yield of

bread in comparison to the control. TGase improves protein/water

binding capacity and inhibits moisture loss of bread during the baking

process. Bread's yield is related to its moisture content after baking.

Moore et al. observed the same results by addition of TGase at 0–10

t
u/g pro levels to the flour mixture containing skim powdered milk [14].

ip
Specific volume

cr
As presented in Table 3, addition of guar gum at 20 g kg-1 and 30 g

us
kg-1 levels increased the specific volume of bread in comparison to the

control. The reason is that hydrocolloids (guar gum) increase the

an
viscosity of dough and improve its development and gas holding [19].

Gambus et al. observed the same results by addition of guar gum to

M
the mixture of corn and potato starches [25].

TGase addition decreased significantly the specific volume of bread

d

compared to the control (P<0.05), although one exception was

e

observed in the G3T1 formula with no significant difference to the

pt

control. TGase activity leads to the formation of cross-link binding

ce

between glutamine and lysine and so inhibits the expansion of gas

cells during the fermentation process. As a result, the specific volume

Ac

of bread will decrease by enzyme activation. The formulae containing

more than 10 u/g pro of TGase showed more decrease in specific

volume. Moore et al. observed a significant decrease in bread volume

when they used 10 u/g pro of TGase [14]. The same results were

observed by Basman et al. [26]. Renzetti et al. also found a significant

11
Page 11 of 23
 decrease in the volume of breads made by brown rice and black wheat

fortified with 10 u/g pro of Tgase [27].

Hardness

The results are shown at Table 4. Addition of 20 g kg-1 and 30 g kg-1

guar gum decreased significantly crumb hardness on the baking day in

t
comparison to the control (P<0.05). Mezaize et al. found similar results

ip
when adding xanthan and guar gums to the mixture including rice flour,

cr
corn starch, corn flour and potato starch [12]. They concluded that

us
hydrocolloids cause the softer texture in gluten-free formulations.

Onyango et al. reported that HPMC has the least effect on the
an
firmness of gluten-free formulations [28]. They also represented that

firmness is closely related to hydrocolloid concentration. TGase

M
addition at 1 and 10 u/g pro levels did not have any significant effect

on the crumb hardness of gluten-free formulae comparing to the

d

control, except to G2T10, which increased the crumb hardness by

e

addition of 10 u/g pro of TGase in comparison to the control or the

pt

formulae containing 1 u/g pro of TGase. Generally, the formulae

ce

containing 10 u/g pro of TGase and lower levels of guar gum (20 g kg-
1
) had the most crumb hardness.
Ac

Increasing the bread’s crumb hardness by TGase addition has been

studied earlier in wheat [21], gluten-free breads [14], and rye dough

[29]. Renzetti et al. also obtained similar results by adding TGase to

flours made from brown rice and black wheat [27]. Crumb hardness

increasing in non-gluten bread compared with controls with

undesirable texture could be considered as an act of a texture

12
Page 12 of 23
 modifier. Crumb hardness is developed by cross-linking proteins.

Addition of 10 u/g pro TGase improves protein phase and forms

protein network with smaller pores followed by formation of decreased-

volume breads. Specific volume is adversely related to crumb

hardness so that increasing of crumb hardness leads to decrease in

t
bread volume [5,14,30–32].

ip
Staling

cr
As shown in Table 4, crumb hardness of manufactured breads

us
increased during 24, 48 and 72 h of storage in all formulae that

represented the improvement of staling. The studies by Moore et al.

an
[14], Gambus et al. [25], Onyango et al. [28] and Krupa-Kozak et al.

[33] also gave similar results. In our study, harness increase on 24 h of

M
storage and was significant in the formulae containing 20 g kg-1 guar

gum (P<0.05), but 30 g kg-1 addition of guar gum did not show any
d

significant difference. Due to undesirable texture of the control, its

e

texture was disrupted before achieving the optimal level of

pt

compression on 0 h of storage, indicating that it has lost its

ce

resistance/flexibility to any change.

On 48 h, the formulae containing 20 g kg-1 guar gum without TGase

Ac

and with 1 u/g pro of TGase as well as the formulae containing 30 g

kg-1 guar gum without TGase could not be analyzed by texture

analyzer because of over-staling and shrinkage of the breads exposed

to compression. Crumb hardness increased in the treated formulae on

48 h of storage. There were no significant differences in crumb

13
Page 13 of 23
 hardness in the formulae containing 30 g kg-1 guar gum and 10 u/g pro

of TGase due to enzyme effect on staling retardation.

On 72 h of storage, the formulae containing no TGase and 1 u/g pro

of TGase did not have desired stability to be analyzed by texture

analyzer because of the gas cells, over-growth, thinning of the gas

t
cells’ walls, reduced resistance to deformation and reduced moisture

ip
content in the bread crumb (due to moisture migration from crumb to

cr
crust). But the formulae containing 10 u/g pro of TGase did not show

us
significant difference in crumb hardness after 72 h of storage; this

indicates that staling was delayed owing to enzymatic activity. Also,

an
moisture content decreased smoothly in these formulae during the

time and there were no significant differences on 72 h in comparison to

M
24 h of storage. Lower staling retardation in these formulae was

probably due to the formation of protein network by Tgase, which traps

d

water and inhibits its migration from crumb to crust; so bread staling is
e

delayed in these formulae as compared to the formulae without any

pt

protein network or those containing weak protein network [21].

ce

Sensory evaluation

Table 5 shows the effect of different concentrations of guar gum and

Ac

TGase on the formulae’s sensory attributes. The results showed that

addition of guar gam at two levels, 20 g kg-1 and 30 g kg-1, improved

the total score of sensory attributes compared to the control.

TGase addition at 1 u/g pro level had resulted in better total score

the treated formulae comparing to the control, but addition of 10 u/g

pro of TGase led to lower sensory score comparing to the control,

14
Page 14 of 23
 formulae TGase-free and those containing lower TGase contents. This

result can be attributed to appreciable reduced volume and condensed

crumb of the bread samples. The most total scores were belonged to

G2T0 and G3T1, and the formulae containing 10 u/g pro of TGase had

the least sensory scores so that they were rejected by our sensory

t
evaluators.

ip
Conclusions

cr
The findings of this work showed that addition of guar gum and

us
microbial TGase enzyme resulted in the higher stability against the

an
mixing process. It was indicated that formed protein network was the

same as gluten structure, and consequently the dough texture was

M
improved. Adding 10 u/g pro of TGase had the highest positive effects

on dough yield and moisture maintenance of the breads after the

d

baking process and consequently, increased the breads’ yield.

e

TGase had negative effect on the specific volume of breads at both

pt

levels; however, guar gam had positive role in enhancing their specific
ce

volume. Guar gum addition alone did not have any effect on the

breads’ texture, but its simultaneous incorporation by TGase at level 1

Ac

u/g pro had positive effect and at 10 u/g pro had negative effect on

their texture. Guar gum addition resulted in texture softening on the

production date, but TGase addition and its dose elevation led to more

crumb hardness of the bread samples at this time. Overall, we can

conclude that high quality gluten-free breads could be produced by

addition of 30 g kg-1 of guar gum and 1 u/g pro of TGase enzyme to

the formulation.

15
Page 15 of 23
 Acknowledgements

The data were obtained from the results of an approved research

project (P/25/47/7138) of the National Nutrition and Food Technology

Research Institute, Iran. The authors would like to thank the Cereal

t
Research Center of Ministry of Industry, Mine and Trade of Iran for

ip
provision of the equipment required for measurement.

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us
References

[1] M. Blades, Nut. Food Sci. 97 (1997) 146–151.

an
[2] J.A. Murray, Am. J. Clin. Nut. 69 (1999) 354–365.

[3] B. Shahbazkhani, R. Malekzadeh, M. Sotoudeh, K.F. Moghadam,

M
M. Farhadi, R. Ansari, A. Elahyfar, K. Rostami, Eur. J. Gastroen.

Hepat. 15 (2003) 475–478.

e d

[4] M.M. Niewinski, J. Am. Diet. Assoc. 108 (2008) 661–672.

pt

[5] E. Zannini, E. Pontonio, D.M. Waters, E.K. Arendt, Appl. Microbiol.

Biot. 93 (2012) 473–485.

ce

[6] N.Y. Haboubi, S. Taylor, S. Jones, Postgrad. Med. J. 82 (2006)

672–678.
Ac

[7] A. Lazaridou, D. Duta, M. Papageorgiou, N. Belc, C.G. Biliaderis, J.

Food Eng. 79 (2007) 1033–1047.

[8] E. Gallagher, T.R. Gormley, E.K. Arendt, J. Food Eng. 56 (2003)

153–161.

[9] M.M. Moore, T.J. Schober, P. Dockery, E.K. Arendt, Cereal Chem.

81 (2004) 567–575.

16
Page 16 of 23
 [10] L. Cato, J. Gan, L. Rafael, D. Small, Food Aust. 56 (2004) 75–78.

[11] D. Ćurić, D. Novotni, D. Tušak, I Bauman, D. Gabrić, Agric.

Conspec. Sci. 72 (2007) 227–232.

[12] S. Mezaize, S. Chevallier, A. Le Bail, M. De Lamballerie, J. Food

Sci. 74 (2009) 140–146.

t
[13] A.A. Anton, S.D. Artfield, Int. J. Food Sci. Nutr. 59 (2008) 11–23.

ip
[14] M.M. Moore, M. Heinbockel, P. Dockery, H.M. Ulmer, E.K. Arendt,

cr
Cereal Chem. 83 (2006) 28–36.

us
[15] AACC International, Approved Methods of the American

Association of Cereal Chemists, 10th ed., The Association, St.

Paul, MN, 2000. an

[16] M. Mohammadi, N. Sadeghnia, M.H. Azizi, T.R. Neyestani, A.M.
M
Mortazavian, J. Ind. Eng. Chem. 20 (2014) 1812–1818.

[17] C. Marco, C.M. Rosell, J. Food Eng. 88 (2008) 94–103.

d

[18] J.S. Wang, M.M. Zhao, X.Q. Yang, Y.M. Jiang, C. Chun, Food
e

Hydrocolloids. 21 (2007) 174–179.

pt

[19] C.M. Rosell, M. Haros, C. Escrivá, C. Benedito De Barber, J. Agr.

ce

Food Chem. 49 (2001) 2973–2977.

[20] A. Houben, A. Höchstötter, T. Becker, Eur. Food Res. Technol.

Ac

235 (2012) 195–208.

[21] J.A. Gerrard, S.E. Fayle, A.J. Wilson, M.P. Newberry, M. Ross, S.

Kavale, J. Food Sci. 63 (1998) 472–475.

[22] H.D. Belitz, W. Grosch, Food Chemistry, 2nd ed., Springer, 1999.

[23] C. Kuraishi, K. Yamazaki, Y. Susa, Food Rev. Int. 17 (2001) 221–

246.

17
Page 17 of 23
 [24] P.C.H.R. Lorenzen, H. Neve, A. Mautner, E. Schlimme, Int. J.

Dairy Technol. 55 (2002) 152–157.

[25] H. Gambus, M. Sikora, R. Ziobro, Acta Scitiarum Polonorum. 6

(2007) 61–74.

[26] A. Basman, H. Köksel, P.K.W. Ng, Eur. Food Res. Technol. 215

t
(2002) 419–424.

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[27] S. Renzetti, F. Dal Bello, E.K. Arendt, J. Cereal Sci. 48 (2008) 33–

cr
45.

us
[28] C. Onyango, G. Unbehend, M.G. Lindhauer, Food Res. Int. 42

(2009) 949–955.
an
[29] M. Beck, M. Jekle, P.L. Selmair, P. Koehler, J. Cereal Sci. 54

(2011) 29–36.
M
[30] M. Gómez, B. Oliete, C. Rosell, V. Pando, E. Fernandez, Food

Sci. Technol. 41 (2008) 1701–1709.

d

[31] D. Sabanis, C. Tzia, Food Sci. Technol. Int. 17 (2011) 279–291.

e

[32] B. Miñarro, E. Albanell, N. Aguilar, B. Guamis, M. Capellas, J.

pt

Cereal Sci. 56 (2012) 476–481.

ce

[33] U. Krupa-Kozak, R. Altamirano-Fortoul, M. Wronkowska, C.M.

Rosell, Eur. Food Res. Technol. 235 (2012) 545–554.

Ac

18
Page 18 of 23
 Table 1
Quantities of guar gum and TGase used in the
formulations of gluten-free breads.

Formulaa Guar (g kg-1)B TGase (u/g pro)C

ControlA 0 0

t
G2T0 20 0

ip
G2T1 20 1

cr
G2T10 20 10

us
G3T0 30 0

G3T1 30 1

G3T10
a
30
an 10

Control (pan bread without guar gum and TGase).

M
Gluten-free breads were prepared according to control
manner and materials but with the addition of guar gum
and TGase.
d

b
Guar gum was added as g kg-1 of total flour basis
e

(including: rice flour, soy flour and corn starch).

pt

c
TGase was added as unit per gram of basic protein at
each formula.
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Page 19 of 23
Table 2
Effect of different levels of guar gum and TGase on the farinograph properties
of gluten-free breads.
Dough Dough
Formulaa Water Degree of
development resistance
absorption (%) softening (BU)
time (min) (min)

t
157.5 ± 3.53b

ip
Control 48.35 ± 0.49a 2.7 ± 0.28abc 1.6 ± 0.14ab

G2T0 59.45 ± 1.62bc 2.35 ± 0.07abc 2.00 ± 0.56b 84.5 ± 14.85a

cr
G2T1 60.55 ± 0.07bc 2.10 ± 0.14ab 2.00 ± 0.28b 63.00 ± 1.41a

us
G2T10 59.8 ± 0.42bc 1.85 ± 0.49a 2.1 ± 0.00b 85.00 ± 21.21a

G3T0 62.65 ± 0.63c 2.85 ± 0.21bc 2.15 ± 0.07b 76.5 ± 3.53a

G3T1 62.45 ± 0.35c

an
3.10 ± 0.56c 2.00 ± 0.28b 67.5 ± 3.53a

G3T10 60.45 ± 1.06bc 3.05 ± 0.07bc 2.20 ± 0.28b 61.5 ± 2.12a

M
a
For formula descriptions see Table 1.
Values are mean±standard deviation (SD) of duplicate treatments (six
d

repetitions for each formula). Means followed by different letters within a

e

column are significantly different (P<0.05).

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Page 20 of 23
Table 3
Effect of different levels of guar gum and TGase on the quality attributes of gluten-free
breads.

Formulaa Specific volume

Moisture (g kg-1) Dough yield (%) Bread yield (%)
(cm3/g)
Control 368.9 ± 13.7a 156.17 ± 0.24a 145.17 ± 0.63a 1.76 ± 0.03cd

t
G2T0 433.6 ± 0.7bc 169.89 ± 0.48d 150.95 ± 1.20b 2.08 ± 0.04f

ip
G2T1 428.9 ± 1.7bc 169.82 ± 0.48cd 153.00 ± 1.55bcd 1.60 ± 0.03b

cr
G2T10 432.2 ± 2.1bc 166.36 ± 0.48b 156.40 ± 1.96d 1.43 ± 0.03a

us
G3T0 439.5 ± 1.4c 170.79 ± 0.48d 152.50 ± 1.39bc 1.93 ± 0.06e

G3T1 442.5 ± 2.8c 169.77 ± 0.48cd 155.20 ± 1.96cd 1.80 ± 0.02d

G3T10
a
469.6 ± 3.9d 173.91 ± 0.47e
an 162.75 ± 1.44e 1.43 ± 0.05a

For formula descriptions see Table 1.

M
Values are mean±standard deviation (SD) of duplicate treatments (six repetitions for
each formula). Means followed by different letters within a column are significantly
different (P<0.05).
e d
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Page 21 of 23
Table 4
Hardness (N) of gluten-free breads formulated with different levels of guar gum and
TGase.

Hardness (N)
Formulaa
Hour 0 Hour 24 Hour 48 Hour 72
Control 72.47 ± 2.42c – – –

t
ip
G2T0 43.36 ± 3.99b 60.65 ± 7.92ab – –

cr
G2T1 72.83 ± 4.63c 75.90 ± 2.15b – –

G2T10 136.84 ± 5.54d 168.39 ± 15.62c 171.48 ± 18.91b 172.88 ± 1.29b

us
G3T0 26.54 ± 2.37a 37.88 ± 5.63a – –

G3T1 66.13 ± 3.87c 75.17 ± 10.02b 97.98 ± 8.79a –

G3T10 62.30 ± 4.99c 87.15 ± 14.80b

an 100.77 ± 4.84a 107.30 ± 8.96a
a
For formula descriptions see Table 1.
M
Values are mean±standard deviation (SD) of duplicate treatments (six repetitions for
each formula). Means followed by different letters within a column are significantly
d

different (P<0.05).
e
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Page 22 of 23
 Table 5
Sensory characterization of fresh gluten-free breads formulated with
different levels of guar gum and TGase.
Formul Crust Crumb Chewin Overall
Form Texture Odor Taste
aa color color ess score
Cont 7.00 ± 11.1 ± 6.00 ± 10.60 ± 11.00 ± 14.30 ± 10.70 ± 70.70 ±

rol 0.67e 0.87de 0.67b 0.70c 0.82c 0.67c 0.82b 1.63b

t
ip
5.90 ± 12.00 ± 8.30 ± 10.70 ± 13.00 ± 16.20 ± 12.40 ± 78.50 ±
G2T0 cd e d c e d c
1.10 0.94 0.48 0.82 0.67 0.92 0.70 3.34c
4.80 ± 9.40 ± 7.20 ± 10.40 ± 13.20 ± 16.20 ± 11.10 ± 72.30 ±
G2T1

cr
b c c c e d b
0.63 1.17 0.79 1.35 0.63 0.79 0.87 2.83b

G2T1 3.60 ± 6.60 ± 4.60 ± 6.60 ± 9.20 ± 11.50 ± 8.80 ± 50.90 ±

us
0.70a 0.70b 1.26a 0.85a 0.92a 1.08a 1.23a 2.51a
0

5.60 ± 0.60 ± 7.40 ± 8.70 ± 11.80 ± 16.30 ± 10.70 ± 71.10 ±

G3T0 bc cd c b cd d b
1.07 1.17 0.84 1.34 1.14 0.82 1.89 4.46b

G3T1
6.50 ±
1.08cde
11.60 ±
1.26de
7.20 ±
0.63c
11.80 ±
1.03d
an
12.50 ±
0.53de
16.50 ±
0.53d
10.90 ±
0.87b
77.00 ±
2.90c

G3T1 3.90 ± 4.80 ± 4.70 ± 8.50 ± 9.70 ± 13.00 ± 9.20 ± 53.80 ±

M
0.74a 0.63a 0.82a 0.85b 0.48ab 1.05b 1.32a 2.20a
0
a
For formula descriptions see Table 1.
Means followed by different letters within a column are
d

significantly different (P<0.05).

e
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Page 23 of 23