Microstructure changes

during baking
Advances in Molecular Structuring of Food Materials

Prof. Carmen Tadini

University of São Paulo
NAPAN – Food and Nutrition Research Center

1
1. Importance and general aspects

Bread in its many forms is one of the

most staple foods consumed.

2
1. Importance and general aspects

It is important to understand about the

unique properties presented by the
wheat proteins, namely their ability to
form a cohesive mass of dough.

3
2. Breadmaking process
To convert wheat flour into an aerated and palatable food

4
 2. Breadmaking process: largely common steps
Mixing of the ingredients according to the formulation

Salt
Yeast
Flour Water
Other
ingredients

Spiral dough mixer

http://www.hiwtc.com/
5
2. Breadmaking process: mixing

Mixing of the ingredients according to the

formulation

 to disperse uniformly the ingredients;

 to dissolve and hydrate the ingredients;
 to contribute energy to the gluten development;
 to incorporate air bubbles within the dough.

6
2. Breadmaking process:
development of the gluten structure

The gluten structure (hydrated proteins) is

developed in the dough through the
application of energy during mixing, often
referred to as “kneading”;

This step is responsible for the

transformation wheat flour and
water into a cohesive dough with
viscoelastic properties;

7
 3. Principles of dough formation: Random
development of the gluten structure orientation

(b)
(c)
(a) If
As
Atthe
mixing
mixing
early proceeds,
stagetime pass
more
the
of mixing, the
protein
optimum becomes
development,
gluten fibrils are hydrated
theand
in contact
the
cross-links
withglutenins
begin
tend
the mixer totobreak;
blade, align,
bowl the
gluten networks
sides and are developed
other flour
by the cross-linking of protein
particles;
with disulphide bonds;

Molecular interpretation of gluten development

(Zaidel et al., 2010) 8
3. Principles of dough formation:
development of the gluten structure

Wheat storage protein has unique properties;

No other cereal storage protein possesses the ability to
form a viscoelastic dough when wetted and kneaded.

In studying dough formation, it is common to observe

the physical events on a macro scale, at the supra-
molecular level;
The macro scale properties of dough change with time
and our knowledge about these modifications in
rudimentary, at best.

9
3. Principles of dough formation:
development of the gluten structure

The real reason for understanding the dough formation is

the final product – a good loaf of bread!

The two characteristics that define “good” dough are:

 The ability to retain gas (CO2);

 A proper balance of viscous flow and elastic
strength.

Gluten is the component of the dough that determines

how well these requirements are met.

10
3. Principles of dough formation:
flour and dough components

Wheat flour components (dry basis) can be

classified into six groups:
 Starch;
 Storage (gluten) proteins;
 Non-starch polysaccharides (pentosans);
 Lipids;
 Water-soluble proteins;
 Inorganic compounds (ash).
11
 3. Principles of dough formation: Starch

The largest portion of flour (76 g/100g d.b.)

 Starch;
17 % (d.b.) amylose
-1,4 linked glucose units
MW =100,000 Da

58 % (d.b.) amylopectin
-1,4 and -1,6 linked glucose units
MW =20,000,000 Da
12
3. Principles of dough formation: Starch

Protein-protein
interaction Starch
granules

Protein
phase
SEM mag 5000 x

13
3. Principles of dough formation:
Protein - flour having 12 % protein contains about
10 % of gluten

Gliadin
single-chain polypeptides
2104 < MW < 7104

Glutenin

multiple-chain polymeric
Mixing proteins

Disulphide
bond formation

14
4. Rheology and bread quality

Within the cereal science community:

 Rheological properties of dough are related
to the bread quality;
 Gluten proteins are responsible for variations
in baking quality;
 In particular, the insoluble fraction of HMW
glutenin polymer is related in dough
strength;
 the exact molecular mechanisms responsible
for this variation still remain unclear;

15
4. Rheology and bread quality
Challenges:

 In polymer physics, it is widely accepted

that molecular size, structure and MWD are
linked to their rheological properties;

 Since gluten can be viewed as a polymer, an

understanding of its rheological properties in
relation to the molecular structure and its
functional behavior during the breadmaking
process is highly desirable;
16
4. Rheology and bread quality
Challenges:

 However, many of the rheological tests

on dough are inappropriate, because
they do not use deformation conditions
appropriate to baking, and are quite
often insensitive to changes in molecular
weight and structure;

17
4. Rheology and bread quality
Rheology is the study of the flow and deformation of materials.

Mixing
behavior

Product
quality Rheology Baking
performance

Mouthfeel
and texture

18
4. Rheology and bread quality
The general aims of rheological measurements are:

 To obtain a quantitative description of the

materials' mechanical properties;
 To obtain information related to the molecular
structure and composition of the material;
 To characterize and simulate the material's
performance during processing and for quality
control.

19
4. Rheology and bread quality
Rheological tests attempt to measure the forces required to
produce given controlled deformations, such as:

Compression Uniaxial extension

Biaxial extension

F [mN]

t [s]
Force-displacement curve 20
4. Rheology and bread quality
m = 3000 kg

 = 30 kPa
A = 1 m2

21
4. Rheology and bread quality

There are many tests methods according to

the type of strain imposed:
 Compression;
 Extension;
 Shear;
 Torsion;
Or depending of the magnitude of the
imposed deformation:
 Small;
 Large;
22
4. Rheology and bread quality

For measuring cereal products, the main

techniques have been divided in:

 Descriptive empirical methods;

 Fundamentals measurement;

Descriptive empirical techniques:

 There are a lot used mainly by cereal

industry in evaluating performance during
processing and for quality control;
23
4. Empirical methods
Farinograph
• Dough
• Mixing time, torque, apparent viscosity

Extensograph
• Dough
• Extensibility

Rapid visco amylograph (RVA)

• Pastes, suspensions
• Apparent viscosity, gelatinization temperature

Though these techniques do not provide data in fundamental

units, important information on the quality and performance
on cereal products are obtained.
24
4. Empirical methods

However, these measurements are

not strictly ‘rheological’ tests since:
 the sample geometry is variable and not well
defined;

 the stress and strain are uncontrolled,

complex and non-uniform; and

 it is not possible to define any rheological

parameters such as stress, strain, strain rate,
modulus or viscosity.

25
4. Empirical methods

Therefore, these tests are purely

descriptive and dependent on:
 the type of instrument;

 size and geometry of the test sample;

and

 the specific conditions.

26
4. Fundamental methods

Oscillatory tests
• Fluids, pastes, batters, dough
• Dynamic shear moduli, dynamic viscosity

Uniaxial extension
• Dough
• Extensional viscosity

Dough inflation system

• Dough
• Strain hardening

These techniques measure well-defined physical properties in

fundamental units.

27
4. Fundamental methods
However, these measurements have many
problems since:
 Complex instrumentation which is expensive, time
consuming and requires high level of technical
skill;
 Often inappropriate deformation conditions;
 Difficulty in interpretation of results; and
 Slip and edge effects during testing.

Therefore, these tests are independent on:

 The type of instrument;
 Size and geometry of the test sample; and
 The specific conditions.

28
 5. Uniaxial extension – empirical method
The required force to deform
the dough is expressed in
Extensograph Units (EU);
From the extensograph load-
extension curve, several
parameters can be derived;

Rmax

Emax
Disadvantage:
The rate of deformation under
fermentation is three orders of Atot

magnitude smaller than the

measured maximum rate.
29
 5. Uniaxial extension – fundamental method
Kieffer dough and gluten extensibility rig

The dough is stretched in one direction;

Again from force-extension curve you can
obtain parameters as stress and strain:

F
L
L L

A
; 
L0
L
 ln  
 L0 
F [N] Maximum
point at
Rmax fracture
Disadvantage:
For the tests, the original sample
dimensions are considered;
But their dimensions change extensively E1

and non-uniformly during testing. Emax Etot d [mm]

30
 5. Uniaxial extension – theory
Samples preparation in a mould:
ten pieces 5 cm long

wherein:
y0: specific point where the deformation is zero [mm]
L0: initial length [mm] v
Lt: length at the time t [mm]
w: width of the gap [mm]

2
w 
L0  2     y02 Lt/2
b1
2
h
2
w 
Lt = 2 ×   +  yt + y0 
2 b2

2

L0/2
y0

31
 5. Uniaxial extension – theory

 2   y  y  
2
 w 2

 Lt  t 0
H  ln    ln  
 L0 
w 2   y
2
 2 
 0 
 
d H 1 dL 1 2  yt  y0  dyt 4  yt  y0 
    v
 2
2
dt Lt dt Lt 2 dt Lt
w   yt  y0 
2

Lt/2
b1

h
b2

L0/2
y0

32
5. Uniaxial extension –
comparison of micro-extensograph with a extensograph

4
H [dimensionless] micro-extensograph

extensograph

0
0 100 200 300 400
d [mm]

Hencky strain (H) as a function of the hook displacement (d).

33
5. Uniaxial extension – force balance

Fm

yt
Fd

 Lt/
2


y0

Forces acting on the dough piece

34
5. Uniaxial extension – Stress calculation

Fm

yt
Fd

 Lt/
2


y0

Forces acting on the dough piece

35
Effect of maize resistance starch (MRS) and transglutaminase (TG)
on rheological properties of pan bread dough – Central Composite Design(22)
0.17

TG1 [g/100g] 0.15

0.10

0.05

0.03
1.5 3.5 8.5 13.5 15.5
1 base on 100 % mixture MRS1 [g/100g] 36
Effect of MRS and TG on rheological properties of pan bread dough –
uniaxial extension

max [kPa]
max [kPa]

37
 6. Biaxial extension – empirical method
During proof and baking the growth
and stability of gas bubbles within
the dough determines the expansion
of the dough and therefore the
ultimate bread volume and texture;

Rupture
point
P [mmH2O]
wherein:
P: maximum pressure [mmH2O]
Bubble
L: extensibility at rupture [mm] pressure
W: deformation energy 10-4 [J]
Bubble
height

0.0 2.5 5.0 7.5 10.0 12.5 15.0

t [s]
38
 6. Biaxial extension – fundamental method
Dough inflation system (DIS)

 The deformation closely resembles practical

conditions experienced by the cell walls around the
expanding gas cells within the dough;

 Researchers advise that these tests are more

relevant for bread making quality than small strain
dynamic measurements.

Parameters Conventional DIS Baking/proofing

tests conditions
Air flow rate (10  2000) mLmin-1
Strain rate (0.01  0.1) s-1 (0.001  0.2) s-1 (0.0001 – 0.001) s-1
Strain (0.1 – 1) % > 100 %
Shear, dynamic oscillation Biaxial extension
39
6. Biaxial extension – theory
Samples preparation in a mould:
five circular samples
55 mm diameter with 8 mm thickness


R

P h

0

40
 6. Biaxial extension – force balance
The internal pressure on a local thickness is:
thin shell of dough
P 1 2
2
 
 r1 r2
And for a near-spherical bubble:

1 1 = 2 = ; r1 = r2 = R;

 P  R
2
1


2 R

r2 P h
r1
r
wherein:
P: internal pressure [mmH2O]
: stress [kPa] 0

41
6. Biaxial extension – force balance

The estimate thickness () is:


R
2
  h  2
 1  2  P h
0  a 
r

That was directly measured by optical

technique. Therefore:
0
 P  R  P   a  h   P   a  h   h 2 
2 2 2 2 2

   1 a
 
2 4h 4h0 
2
a 

 P   a 2  h 2 
3


4h0 a 4
L
dL L A0 1 0  h2 
H    ln  ln  ln  ln 1  2 
L0
L0 L0 A 2   a 
dV
4h
d
  H  2 dt2
dt h  a  
42
6. Biaxial extension

 [mm]

h [mm]

Spherical bubble crown thickness () as a function of the bubble height (h),
(measured); (calculated).

(Tanner et al., 2008)

43
6. Biaxial extension –
If the pressure and bubble height versus time data is
transformed into a stress-strain curve:

Rupture
point
P [mmH2O]

Bubble
pressure

Bubble
height

0.0 2.5 5.0 7.5 10.0 12.5 15.0

t [s]

44
6. Biaxial extension
During large stretching of materials, the cross-sectional area changes in
a non-uniform way. If the pressure and bubble height versus time data
is transformed into a stress-strain curve:

Rupture
point
P [mmH2O]

Bubble
pressure

Bubble
height

0.0 2.5 5.0 7.5 10.0 12.5 15.0

t [s]
Tension – extension curve for the polar region of a bubble
45
6. Biaxial extension
During large stretching of materials, the cross-sectional area changes in
a non-uniform way. If the pressure and bubble height versus time data
is transformed into a stress-strain curve:

L
dF  d  
 L0 
 [kPa]

Bubble
rupture

 [-]
Tension – extension curve for the polar region of a bubble
46
6. Biaxial extension – criterion for instability in tension

The condition of necking instability is:

F  A  0 ; dF  dA  Ad   0
dA d 
and since:   d
A 
d
   at dF  0 
d
L
dF  d  

 [kPa]
 L0 

  k n
Bubble
rupture

 [-]
47
Effect of maize resistance starch (MRS) and transglutaminase (TG)
on rheological properties of pan bread dough – Central Composite Design(22)
0.17

TG1 [g/100g] 0.15

0.10

0.05

0.03
1.5 3.5 8.5 13.5 15.5
1 base on 100 % mixture MRS1 [g/100g] 48
Effect of MRS and TG on rheological properties of pan bread dough –
biaxial extension

 [kPa]  [kPa]

  1.99  0.33  MRS  0.18  TG  0.34  TG 2   0.20

49
 Strain - hardening
From stress-strain data:
  k exp  n H 

  0.0108  exp  2.21  H 

wherein:
: stress [MPa];
H: Hencky strain [-];
k: coefficient related to the dough viscosity [MPa];
n: strain hardening index [-].
Rupture
point
P [mmH2O]
Values of n greater than
1, indicate strain
Bubble
hardening pressure

Bubble
Greater strain hardening height

Greater bubble failure

Greater loaf volume 0.0 2.5 5.0 7.5 10.0 12.5 15.0
t [s]
50
7. Our approach–
Dynamic rheometer – large deformation

Measured parameters in real time:

c: current [mA];
Vred: reducer speed [rpm].

Calculated parameters:

  k1c
vred
v
wherein: k2
: torque [Nm];
v: blade speed [rpm];
k1: constant [24.91 NmmA-1];
k2: constant [3.96].
51
7. Our approach– Dynamic rheometer – large deformation
The instantaneous specific power (Po ) can be calculated:

2vred   rad min N  m 

Po   
60  m  min s kg 
vred   W 
Po   
30  m  kg 
t
E   Po dt
0

wherein:
m: dough mass [kg];
t: total mixing time [s];
E: specific energy [kJ/kg].

52
Effect of three enzymes: transglutaminase (TG), glucoxidase (Gox)
and xylanase (He) on rheological properties of pan bread dough –
Mixture Design of three components

The dough was prepared with a blend of

12.5 g of MRS plus 87.5 g/100g of wheat flour.

From each run, a dough sample was also

submitted to rheofermentometer tests:

53
Parameters obtained by development curve and gas release curve

Maximum
height

Maximum
pressure

The dough strength is not only

dependent on the total work input,
but also on aeration during the
mixing and produced gas bubbles
during fermentation.

54
Parameters obtained by development curve and gas release curve

A new parameter:

Hm
Hm 
z
Vt Vt 0
z ; z0 
tz0 t0
wherein:
z: normalized gas flow rate [-];
Vt: total gas volume [mL];
subscript 0 indicates control

55
Effect of three enzymes: transglutaminase (TG), glucoxidase (Gox)
and xylanase (He) on rheological properties of pan bread dough

Results:

The new parameter, corrected maximum height (H m )

indicated that dough prepared with higher
concentrations of Gox and TG, presented better
baking performance. TG [4 g/100g]
Hm
20
22
24
26
28
30
32

Gox [4 g/100g] TG [0 g/100g] He [4 g/100g]

56
8. Rheology and bread quality
Gluten and dough are most unique from the
point of material science as they have complex
behavior:

 Rheological properties measured to accuracy

are important for product development both
in research or industry;
 The correlation studies on these properties of
the dough with baking performance of the
end product is an example of such appliance;

57
 References
BLOKSMA, A. H. A Calculation Of The Alveograms Of Some Rheological Model
Substances. Cereal Chemistry, n. 34, p. 126-136, 1957.

CAUVAIN, Stanley P. ; YOUNG, Linda S. Technology of Breadmaking. 2nd ed.. New York:
Springer, LLC, 2007.

CHIN, N. L. ; CAMPBELL, G. M. Dough aeration and rheology: Part 1. Effects of mixing

speed and headspace pressure on mechanical development of bread dough. Journal of
the Science of Food and Agriculture, n. 85, p. 2184-2193, 2005a.

CHIN, N. L. ; CAMPBELL, G. M. Dough aeration and rheology: Part 2. Effects of flour

type, mixing speed and total work input on aeration and rheology of bread dough.
Journal of the Science of Food and Agriculture, n. 85, p. 2194-2202, 2005b.

DUNNEWIND, B. ; SLIWINSKI, E. L. ; GROLLE, K. ; VAN VLIET, T. The Kieffer Dough and

Gluten Extensibility Rig – An experimental evaluation. Journal of Texture Studies, n.
34, p. 537-560, 2004.

LAUNAY, B. ; BURÉ, J. ; PRADEN, J. Use of the Chopin Alveographe as a rheological toll

I. Dough Deformation Measurements. Cereal Chemistry, n. 54, p. 1042-1048, 1977.

LAUNAY, B. ; BURÉ, J. Use Of The Chopin Alveographe As A Rheological Toll II.

Properties In Biaxial Extension. Cereal Chemistry, n. 54, p. 1152-1158, 1977.

SANCHEZ, Diana Buchner de Oliveira. Desempenho Reológico e Entálpico da Massa de

Pão com Amido Resistente de Milho e Transglutaminase. SP: USP, 2009, Dissertação
(Mestrado) .
58
 Prof. Carmen Tadini

catadini@usp.br

http://sites.poli.usp.br/pqi/lea/index_e.html

59