Food Chemistry 122 (2010) 495–499

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Bread improvers: Comparison of a range of lipases with a traditional emulsifier

S. Moayedallaie a,*, M. Mirzaei b, J. Paterson a
a
School of Chemical Sciences and Engineering, The University of New South Wales, Sydney, NSW 2052, Australia
b
Shahid Sadoughi University, Yazd 89167-77441, Iran

a r t i c l e i n f o a b s t r a c t

Article history: This study compares three generations of lipase enzymes with the emulsifier, diacetyl tartaric esters of
Received 20 January 2009 mono-glycerides (DATEM), on white wheat flour bread. Baking recipes with addition of DATEM
Received in revised form 7 September 2009 (4500 ppm), Lipopan F-BG (15 ppm), Gryndamyl Exel-16 (115 ppm), Lipopan Xtra-BG (25 ppm) and Lipo-
Accepted 14 October 2009
pan 50-BG (27.5 ppm) were test-baked after 60 and 150 min fermentations, to study their effects on the
baking characteristics of volume, oven rise, crust colour, crumb texture and colour, shelf-life, flavour and
aroma. The enzymes and emulsifier preparation caused significant increase in bread oven rise and spe-
Keywords:
cific volume with the exception of Lipopan 50-BG, which failed to improve loaf volume in the short fer-
Bread
Enzyme
mentation method. There was no significant difference between other lipase enzymes and DATEM as a
Lipase bread volume improver. Increase in fermentation time resulted in increase in volume in all samples,
Emulsifier except for Lipopan-Xtra.
DATEM Ó 2009 Elsevier Ltd. All rights reserved.
Shelf-life
Aroma
Flavour
Texture
Volume

1. Introduction plete replacement of traditionally used volume-improving agents,

such as emulsifiers with enzyme-based volume improvers, is
In the past few decades, demands for longer shelf-life and con- desirable.
sistent quality in baked goods lead to application of a wide range of Lipases have been used over the past two decades, along with
additives in the baking industry. These additives (bread improv- other enzymes and emulsifiers, to improve some characteristics
ers), including emulsifiers, enzymes, soy flour, oxidants and reduc- of baked goods. They hydrolyse triglyceride esters and produce
tants (Kent & Evers, 1994), are essential for improving dough mono- or di-glycerides, glycerol and free fatty acids. Most lipases
machinability (Haros, Rosell, & Benedito, 2002a), reducing resting act only on specific positions in triglycerides (Hasan, Shah, & Ham-
time, and improving baked goods’ shelf-life (Haros, Rosell, & Ben- eed, 2006) and have fungal or bacterial sources (Arpigny & Jaeger,
edito, 2002b; Leon, Duran, & Benedito De Barber, 2002), volume, 1999). Lipases strengthen dough stability (Primo-Martin, Hamer, &
crust colour, crumb whiteness, aroma and flavour (Martinez-Ana- Jongh, 2006) and increase bread volume, texture and shelf-life.
ya, 1996). In the 1990s, the first generation of lipases was introduced to
Diacetyl tartaric esters of mono-glycerides (DATEM) constitute the market. These lipases hydrolyse the ester bond between glyce-
an emulsifier that has been widely used as a bread improver. DA- rides and fatty acids in positions 1 and 3 of triglycerides, producing
TEM improves bread volume and texture and also dough stability free fatty acids and mono-glycerides, and producing more polar
(Lorenz, 1983; Mettler & Seibel, 1993). Compared to emulsifiers lipids in the dough (Spendler, Christiansen, & Frandsen, 2001).
and chemical agents which are detectable in the final loaf, enzymes Martinez-Anaya and Jimenez (1997) studied the effect of a first
become denatured during baking and become undetectable in the generation lipase, Lipopan 50-BG. They found that this lipase
final products. This allows enzymes to be clean label improvers; strengthened the gluten network but that overdosing produced a
they provide bread improving functions without appearing on stiff dough, resulting in a decrease in loaf volume.
the label (Martinez-Anaya & Jimenez, 1997). Thus, partial or com- The second generation lipases work on both polar and non polar
lipids in the wheat flour, producing more polar components, such
as lysolesitin and di-galactosyl monoacyl glycerol (DGMG) than
* Corresponding author. Tel.: +98 351 8239970; fax: +98 351 8239980.
E-mail addresses: [email protected], [email protected] (S.
the first generation lipases (Spendler et al., 2001). Lipopan F-BG,
Moayedallaie). from this generation, has been studied by Primo-Martin et al.

0308-8146/$ - see front matter Ó 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2009.10.033
496 S. Moayedallaie et al. / Food Chemistry 122 (2010) 495–499

(2006); by acting on wheat polar lipids, such phospholipids and Table 1

galactosyl glycerides, this lipase produces more polar lipids than Characteristics of the assessed commercial enzymes and emulsifiers.

do the first generation lipases. Trade name Type Recommended Used Activity
Lipopan Xtra (Novozymes AG, Dittingen, Switzerland) is a third dosage range dosage (KLU/g)
generation, more concentrated lipase which, it is claimed, has a (ppm) (ppm)

better tolerance toward variations in flour type and dosing. Sto- Lipopan Xtra-BG Lipase enzyme 20–30 25 7.2
jceska and Ainsworth (2008) evaluated the effect of Lipopan 3rd generation
Lipopan F-BG Lipase enzyme 10–20 15 25
Xtra-BG on volume, staling and crumb structure of high fibre white 2nd generation
bread. They reported that this enzyme increases expansion of the Lipopan 50-BG Lipase enzyme 5–50 27.5 50
gluten network, increases the wall thickness and decreases cell 1st generation
density, improving these characteristics of high fibre white bread. Grindamyl Exel-16 Lipase enzyme 80–150 115 –
1st generation
It has been claimed that emulsifiers, such as DATEM, are par-
Panodan A2020 DATEM 3000–6000 4500 –
tially or even totally replaceable with lipase enzymes (Christian-
sen, Vind, Borch, & Heldt-Hansen, 2003; Novozymes, 2006; KLU, kilo lipase unit.
Sources: Danisco (2000) and Novozymes (2003, 2006) product sheets.
Spendler et al., 2001). Two reasons why partial or total substitution
of emulsifiers, such as DATEM with lipase enzymes, interest the
baking industry are the storage and transportation difficulties of 2.2. Methods
emulsifiers, such as DATEM, and the comparative cost. DATEM re-
quires refrigeration because of its tendency to cake. Although mak- 2.2.1. Test baking
ing a gel from DATEM solves the caking problem, the performance The baking recipe of this study included 1000 g of white wheat
of DATEM gel decreases during storage (Azizi & Rao, 2005). Fur- flour, 20 g of salt, 650 g of water, 10 g of instant dry yeast, 30 g of
thermore, the weight potency of DATEM emulsifier is usually1/10 sucrose, 16 g of malt flour and 500 ppm ascorbic acid. Enzymes and
or less of that of lipase (Danisco, 2000; Novozymes, 2003), which emulsifier were added to this recipe as summarised in Table 1.
increases the cost of transport. Short and long fermentation times were used. The short fer-
Because introduction of lipase enzymes, as a bread improver, is mentation method was adopted from the George Weston method
relatively recent, there have been only a few investigations of this for flour quality control. This is an established method that is used
category of baking enzymes. The new generation lipases entered in some major commercial bakeries in Australia, as described in
the market only recently, requiring more investigation of their detail below. The long fermentation method in this experiment
application in baking and their effects on various characteristics was adopted from the AACC method (AACC, 2000a).
of bread. This study compares different generations of lipases, ob- Bread samples were made using the straight dough procedure.
tained from major producers, with DATEM and their effects on Freshly opened yeast was used at the beginning of each day of
bread crumb texture, crust colour, volume, shelf-life, flavour and the baking trials. Dry ingredients were mixed in a Duplex Morton
aroma. mixer (Morton-Mixers, Scotland, UK) for 20 s. Water was added,
followed by 30 s of mixing at low speed. The dough was then
scratched down from the mixer edges and mixed for 3 min at high
2. Materials and methods speed. The water and water jacket temperatures were adjusted to
give a dough temperature of 29 ± 0.5 °C after mixing. As the Mor-
2.1. Materials ton mixer mixes differently from the farinograph mixer, the opti-
mum water content of 65% was adjusted for the Morton mixer.
2.1.1. Flour The doughs were cut to two 350 g pop loaves and one 750 g sand-
The flour was a commercial white wheat flour provided by wich bread. They were then moulded and rested in a cabinet at
George Weston Milling (George Weston Milling Ltd., Sydney, Aus- room temperature for 8 min. In the short fermentation method,
tralia). This flour has 11.5% protein with Farinograph water absorp- the dough pieces were placed into a prover at 40 °C and 80% rela-
tion of 68.8% at 500 Brabender units (Brabender GmbH, Duisburg, tive humidity until the height of the 350 g dough in the tins
Germany). reached 10.3 cm. This was a 60 ± 1 min fermentation period for
all samples. In the long fermentation method, dough was fer-
2.1.2. Improvers mented at 30 °C to the same height for 150 ± 1 min and the dough
The four lipase enzymes were of commercial quality. Lipopan F- was punched after 42 and 87 min of fermentation. Breads were
BG, Lipopan Xtra-BG and Lipopan 50-BG were supplied by Novo- baked in an electric oven at 210 °C for 30 min.
zymes Company (Novozymes AG, Dittingen, Switzerland) and
Gryndamyl Exel-16 by Danisco Cultur Company (Dansico Cultor 2.2.2. Bread analysis
A/S, Braband, Denmark). These two companies are the main pro- Bread volume, crust colour, crumb texture and colour, oven rise
ducers of bakery enzymes and these lipases are used worldwide. and flavour and aroma of the end-product were assessed. Oven rise
Lipopan 50-BG and Gryndamyl Exel-16 are first generation lipases. was assessed by measuring the average height difference of the
Lipopan F-BG is second generation and Lipopan-Xtra is third gener- 350 g pop loaves in the tins just before and after putting them into
ation. DATEM emulsifier, Panodan A2020, is produced by Danisco the oven. The loaves were cooled and volume and specific volume
Cultur and is a mixture of DATEM (80%) with calcium carbonate of pop loaves were measured using the seed replacement process,
(20%) as a neutral anti-caking agent. In this study, mid-value of according to the AACC method (AACC, 2000b).
the dosage range of each enzyme, recommended by its manufac- The crumb brightness of baked samples was measured using a
turer, was added to the recipe (Table 1). The control contained Konica Minolta Spectrophotometer (Konica Minolta Ltd., Osaka, Ja-
no added lipase or DATEM. pan), and using the L*a*b* system. The value of L* b*
(lightness yellowness/blueness), which is used as the brightness
2.1.3. Yeast indicator, was used to compare the crumb colour difference be-
Instant dry yeast (FermipanTM from Gist-brocades, Delft, Nether- tween the samples. Crust darkness was determined by value of
lands) was the fermentation agent. Full fat enzyme-active soy flour 100 L* (100 L* = 0 corresponds to white and 100 L* = 100 cor-
and malt flour were used as conditioners in all trials of the study. responds to black) (Sahlstrom & Brathen, 1996).
 S. Moayedallaie et al. / Food Chemistry 122 (2010) 495–499 497

Crumb firmness was determined on the same loaves according trol (P < 0.05) except for Lipopan 50-BG in the short method
to AACC method (AACC, 2000c), using a texture analyser, model (Table 2).
TA-XT2i (Stable Microsystems, Surrey, U.K), with a 25 mm probe. Increase in fermentation time increased the specific volume of
Bread loaves were bagged after cooling for 90 min and kept at all samples except for those containing Lipopan Xtra-BG.
room temperature for one day. The crumb firmness test was per-
formed on the first day after baking. The crust and first two slices 3.2. Colour
of bread were discarded. The maximum force (N) at 1 mm/s speed
and 40% compression of two bread slices (2.5 cm thickness) was Addition of all improvers resulted in significant increase of the
defined as firmness. Eight readings were done for each loaf of crumb brightness compared to the control (P < 0.05), except for
bread. DATEM in the short method. There was no significant difference
A sensory test, with a panel of six professional baking experts, in the brightness between the treated samples. Longer fermenta-
was conducted to assess aroma and flavour of breads made with tion resulted in darker crumb colour. There were no significant dif-
different mixtures of enzymes and emulsifiers. They answered ferences among the samples’ crust darkness values (P < 0.05)
the questions by rating the features of bread sensory quality on (Table 3).
the scale from 1 to 10, where 1 stands for extremely unpleasant
or weak and 10 for extremely pleasant or strong. 3.3. Crumb texture

2.3. Statistical analysis Only DATEM and Lipopan Xtra-BG, in the short fermentation
method, significantly reduced the firmness (P < 0.05) (Table 3).
A table of random numbers (GraphPad software Inc.;
www.graphpad.com) was used to select the order in which sam- 3.4. Sensory evaluation
ples were baked to minimise bias.
The data were analysed using SAS statistical package version 9.1 Among the samples, only Lipopan F-BG and Lipopan Xtra-BG
(SAS Institute Inc., Cary, NC. USA). Tukey’s test was performed to did not show a more significantly intense flavour than the control.
determine minimum significant differences between mean of con- There were no differences among the overall flavour intensities of
trol and mean of other samples and P values less than 0.05 were the samples (P < 0.05).
considered statistically significant. All the baking trials were per- Only Lipopan Xtra-BG and Lipopan 50-BG gave significantly
formed in duplicate. (P < 0.05) less intense acidic flavour than did DATEM. There were
no significant differences of soapy flavour intensities, overall fla-
3. Results vour desirabilities and undesirable aromas between the samples
and the control or among the samples (Table 4).
3.1. Volume
4. Discussion
Oven rise followed the same pattern as did volume and specific
volume, in all samples. Addition of all bread improvers signifi- Compared with the control, all improvers increased specific vol-
cantly increased the bread specific volume compared with the con- ume and oven spring but were not statistically different from one

Table 2
Effect of bread improvers and proof time on oven rise, bread volume and specific volume.

Bread Characteristics Enzyme preparations

Control Lipopan Xtra-BG Lipopan F-BG Lipopan 50-BG Grindamyl Exel-16 DATEM
Proof time 60 ± 1 min
Oven rise (mm)  x ± SD 1.4 ± 0.98 3.2 ± 0.19* 2.9 ± 0.31* 2.9 ± 0.70* 3.1 ± 0.10* 3.2 ± 0.06*
Volume (ml)  x ± SD 1070 ± 96 1223 ± 81 * 1210 ± 88* 1156 ± 60 1193 ± 43* 1236 ± 69*
Specific Volume (ml/g)  x ± SD 3.6 ± 0.41 4.3 ± 0.24* 4.1 ± 0.30* 3.9 ± 0.18 4.1 ± 0.14* 4.2 ± 0.22*
Proof time 150 ± 1 min
Oven rise (mm)  x ± SD 2.2 ± 0.59 3.7 ± 0.29* 4.4 ± 0.22* 4.0 ± 0.22* 3.9 ± 0.25* 4.4 ± 0.22*
Volume (ml) x ± SD 1196 ± 42 1255 ± 71 1319 ± 50* 1278 ± 26* 1284 ± 36* 1333 ± 22*
Specific Volume (ml/g)  x ± SD 4.0 ± 0.19 4.3 ± 0.25* 4.5 ± 0.18* 4.4 ± 0.11* 4.4 ± 0.12* 4.5 ± 0.09*
*
Significantly different from control (P < 0.05).

Table 3
Effect of different bread improvers and varying proof times on crumb colour, crust colour and texture.

Bread characteristics Enzyme preparations

Control Lipopan Xtra-BG Lipopan F-BG Lipopan 50-BG Grindamyl Exel-16 DATEM
Proof time 60 ± 1 min
Crumb coloura  x ± SD 63.2 ± 0.77 67.3 ± 0.33* 67.2 ± 0.86* 68.3 ± 0.17* 66.7 ± 0.75* 65.8 ± 1.62
Crust colourb 
x ± SD 58.7 ± 3.04 59.7 ± 2.51 56.2 ± 4.06 58.9 ± 5.09 58.5 ± 2.73 58.6 ± 5.25
Texture (N) 
x ± SD 6.0 ± 1.3 3.9* ± 0.8 4.2 ± 0.8 4.3 ± 0.7 4.2 ± 0.5 4.0* ± 1.1
Proof time 150 ± 1 min
Crumb coloura  x ± SD 59.5 ± 0.88 64.0 ± 0.44* 64.1 ± 0.15* 70.8 ± 0.43* 63.0 ± 0.72* 63.5 ± 0.95*
Crust colourb 
x ± SD 60.4 ± 4.07 57.6 ± 2.93 56.7 ± 4.51 60.3 ± 3.05 58.2 ± 5.08 58.9 ± 5.75
Texture (N) 
x ± SD 5.0 ± 1.3 3.9 ± 0.8 3.6 ± 1.1 4.0 ± 1.1 3.8 ± 0.9 3.8 ± 0.8
*
Significantly different from control (P < 0.05).
a
(L* b*).
b
(100 L*), N: maximum force (Newton).
498 S. Moayedallaie et al. / Food Chemistry 122 (2010) 495–499

Table 4
Effect of bread improvers on flavour and aroma.

Bread characteristics Enzyme preparations

Control Lipopan Xtra-BG Lipopan F-BG Lipopan 50-BG Grindamyl Exel-16 DATEM
Overall flavour intensity x ± SD 2.1 ± 0.85 3.5 ± 2.45 3.8 ± 2.30 6.8 ± 0.76a 6.8 ± 0.2 1a 4.8 ± 2.70a
Acidic flavour intensity 
x ± SD 4.6 ± 0.31 3.5 ± 2.76b 4 ± 2.10 2.2 ± 1.33b 4.3 ± 2.93 5.7 ± 2.76
Soapy flavour intensity x ± SD 3.5 ± 2.57 5.5 ± 3.79 3.2 ± 1.06 6.1 ± 1.33 2.7 ± 1.10 4.6 ± 2.66
Overall flavour desirability x ± SD 4.2 ± 1.48 5.7 ± 2.76 5.7 ± 0.64 6.2 ± 1.91 4.6 ± 0.42 6.3 ± 0.99
Undesirable aroma x ± SD 6.0 ± 3.89 5.6 ± 2.05 6 ± 1.70 6.9 ± 2.05 4 ± 0.99 4.2 ± 2.83
a
Significantly different from control (P < 0.05).
b
Significantly different from DATEM (P < 0.05).

another except for Lipopan 50-BG in short fermentation, where the 5. Conclusions
specific volume was significantly smaller than the others.
DATEM performed best in all categories except crumb bright- Lipase enzymes and DATEM improved most aspects of bread
ness. The good performance of DATEM, in 60 and 150 min fermen- quality. In shorter fermentation times, DATEM, Lipopan F-BG and
tation times, is not consistent with the study by Gomez et al. Lipopan Xtra-BG were more effective. In longer fermentations, un-
(2004) who found that, after 1.5 h of fermentation, DATEM-treated like the third generation lipase, Lipopan Xtra-BG, moderate
samples showed lower volumes than did the control. This was ex- amounts of the second generation lipase, Lipopan F-BG, signifi-
plained by proofing retardation by emulsifiers (Gomez et al., 2004). cantly increased the bread volume.
Lipopan Xtra-BG had contradictory performances in different
methods. In the short fermentation method, it produced the high-
est specific volume. Good performance of this lipase in short fer- Conflict of interest
mentation is in agreement with results from Stojceska and
Ainsworth (2008), who found that 25 ppm of Lipopan Xtra-BG, in None.
a 55 min fermentation time, significantly increased the bread vol-
ume. In the long fermentation method, Lipopan Xtra-BG’s volume Acknowledgments
decreased remarkably. This could be explained by longer action
of this strong lipase on the dough, producing stiffer dough in the The authors acknowledge George Weston Technologies for pro-
long fermentation method, which leads to decrease in bread vol- viding laboratory equipment and ingredients and Mr. Jeffrey Parker
ume. Like all other research done on bread improvers, we used for his comments on the design and interpretation of the study. We
the range recommended by suppliers’ application sheets as a guide thank Professor A. Stewart Truswell of the University of Sydney for
and treated all the enzymes and emulsifiers in the same way. critical reading of this paper and Behnan Kereshmeh Co. of Yazd for
Brown crust in bread is a result of non-enzymatic browning technical support.
reaction (Maillard type) between amino acids and reducing sugars
(Kent & Evers, 1994). Unlike a-amylase, which produces more su-
gar in the dough, resulting in crust darkness increase, lipases do References
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