Bread Production

By Connor Thornton
Spring 2014

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U21655345

Since Neolithic time humans have used yeasts and bacteria to aid in
food production and longevity. Even without the knowledge of the existence
of microorganisms the processes were possible and reproducible. One of the
most significant of which was the production of bread from wheat and water.
Though bread dough production is traditionally attributed to Saccharomyces
cerevisiae there are a number of different yeasts that can be used as well as
lactic acid bacteria which lead to an assortment of different styles and
flavors of bread.
Bread was revealed as flour-water mixtures tended to collect airborne
yeast spores creating dough. The required microorganisms for dough
formation are abundant in nature and they or their spores are readily
transported by wind. Location is important though as the species of
microorganisms in an area depend on the variety of wheat in the surrounding
areas, and as such will metabolize at different rates producing uniquely
regional breads and flavors (Minervini et al. 2012). A mixture of flour, water
and yeast quickly thickens and becomes dough, yellow and thick.
Sourdoughs are those with a complex community of yeasts and lactic acid
bacteria.
Dough grows over time and will continue to grow until it either runs out
of water or nutrients, or else baked, becoming bread. At any point dough
may be taken and added to another flour-water mixture to produce more
bread this process is known as back slopping. This is extremely common in

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sourdough recipes as the keeping the same yeast and bacteria from batch to
batch becomes exceedingly difficult. Industrially, a post-fermented culture is
taken and used to inoculate the next batch; the new inoculum is then given
salt to alter the yeasts metabolism to produce tastier bread. Back sloppings
are incubated for 6 to 15 minutes at around 25 degrees Celsius. Inoculums
can either be used as a leavening agent to encourage CO2 production or as a
baking improver which improve the quality, the enzymatic activity and/or the
physical and chemical resistance of a culture (Minervini et al. 2012). The
main drawback of back slopping is the increased risk of viral contamination.
Yeast used in bread making may be used to make a variety of different
products including (S)-acetoin (LIU Pei-hai et al, 2013), Ethanol in the
production of biofuels (Neilsen J et al, 2013), acetalaldehyde, amylases,
proteases producing peptides and amino acids, lipases (Antonia, MartinzeAnaya, 1996), xylanases and oxidases (Whitehurst and Oort, 2010) vitamins
B1, B4, panthothenic acid, niacin, folic acid and biotin (Minervini et al. 2012).
Industrially S. cerevisiae and other yeasts can be difficult to work with. Many
yeasts undergo a process called the Crabtree effect where glucose is
degraded aerobically even when it is abundant, disregarding the Pasteur
Effect. This is because the elevated presence of glucose inactivates
cytochrome synthesis (Deken, 1996). Also, acetalaldehyde and ethanol are
difficult to make in high concentrations as they are toxic and will kill the
yeast if the toxins become too prevalent (Antonia, Martinze-Anaya, 1996). In
addition, protein biomass is produced from S. cerevisiae due to the high cell

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production rate and relatively simple pathways. Though, difficulties arise as

S. cerevisiae needs to be fed additional carbon, such as molasses,
intermittently after it has peaked in production in order to prevent catabolite
repression due to the high rate of glucose utilization caused by the Crabtree
effect. Secondly, S. cerevisiae requires a fully aerobic environment to
maximize yield. This second point makes testing difficult as fully aerobic
environments are difficult to make on a small, testing, scale (E. Durao et al.
2013).
As with any product the process of transferring the product from origin
to market takes a large amount of time and the shelf life of the product is
key to preventing losses. In bread the two factors to look out for are staling
and fungal production. Staling is the the hardening and drying of bread over
time and is caused by starch isomerization called retrogradation (Whitehurst
and Oort, 2010). Wheat flour comes from the grinding up of the endosperm
of grain which consists mostly of the starches amylose, a spiral polymer of
glucose, and amylopectin, a branched polymer of glucose (Fredriksson et al,
1998). When heated the polymers gelatinize and isomerizes and become
more difficult to degrade thus resulting in an unappetizing flavor and texture.
When heated the amylopectin recrystalizes forming starch networks that
cause water to be pushed into pockets, unable to move (Goesaert et al,
2009). To combat staling emulsifiers and enzymes are used to prevent
isomerization (Whitehurst and Oort, 2010), as well as maltogenic -amylase

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producing Bacillus stearothermophilus

which degrade amylopectin isomers
(Goesaert et al, 2009).
Airborne microorganisms used for
making bread vary from region to
region. Many regions throughout
Europe produced unique bread with tastes differing from town to town. The
Protected Designation of Origin law enacted by the European Union
effectively allows products to be named from the region they were produced
and banning the use of that name outside said region. This law became a
cornerstone to the bread industry as the uniqueness of bread improved as
effective patents on natural airborne cultures were formed. The United
States saw a similar event as San Francisco sourdough named its primary
yeast Lactobacillus sanfranciscensis. Sourdough is the most common natural
starter in industrial bread production and is used in a majority of traditional
breads. The intertwined relationship of the S. cerevisiae, other yeast sp and
lactobacillus sp. causes a symbiotic relationship where one protects the
other from physical and chemical harm. The bacteria present in various
traditional sourdough cultures include Leuconostoc citreum, Leuconostoc
mesenteroides subsp. mesenteroides, Lactobacillus sakei, L.
sanfranciscensis, , L. plantarum, L. spicheri, L. brevis, L. paralimentarius, L.
casei, L. namurensis, L. gallinarum, L. rossiae, Lactococcus lactis,
Pediococcus pentosaceus P. inopinatus, P. argentinicus, Weissella cibaria, W.

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confusa, W. paramesenteroides, , and Enterococcus durans (Minervini et al.

2012). Lactic acid bacteria can be in numbers greater than 108 CFU/g-1 over
ten times the amount of yeast but in certain regional sourdoughs they have
been numbered at ten thousand to one and others one to one lactic acid
bacteria to yeast.
While there are numerous micro organisms in sourdough the most
common are Candida kruei, C. milleri, Kazachstania exigua, Picha
norvegensis, Saccharomyces cerevisiae, Torulaspora delbrueckii,
Wickerhamomyces anomalus. Flours are made from one of two species of
grain, either T. aesticum, common wheat or T. durum, an artificially selected
wheat grown since ancient time. Sourdoughs made from one wheat use
different cultures than those from the other, likely because the microbes
pathways have differentiated over time. Flour variations also vary the
amount of maltose, glucose, fructose and amino acids in the bread. The pH
of a sourdough culture ranges from 3.7 to 4.3. and consist of .6% maltose, .
5% glucose .4% fructose and .95 g kg-1 free amino acids with T. duram flours
while the T aestivum sourdoughs consist of .2%, .25%, .25% and .6g kg-1 of
maltose, glucose, fructose and free amino acids respectively. The T. durum
sourdoughs contained more than ten times the amount of most amino acids
than with T. aesticum flours (Minervini et al. 2012). D,L lactic acid from the
lactic acid bacteria is the primary cause of pH decrease. The presence of
each compound will decrease during baking by around a 15% leading to less
impact on the finished product (Black et al. 2013). The individual

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concentrations of amino acids will vary

widely between each specific culture and
impact the overall flavor discrepancy
between breads.

Interestingly, T. duram flour

cultures tend to contain a high amount of obligate heterofermentative
lactobacilli and a low density of yeasts and other lactobacilli. Overall,
sourdough cultures can be split into three groups: ones consisting primarily
of homofermentors, ones consisting primarily of facultative
heterofermentative lactic acid bacteria and high yeast concentration, and
those containing primarily obligate heterofermentative lactic acid bacteria
and large amounts of acetic acid as a result. This is supported by phylogenic
data which suggests that the T. aesticum utilizing lactic acid bacteria to be
distantly related to the T. durum bacteria and that the trait for hetero- or
homofermentation was a primary splitting point in lactic acid bacterial
ancestral history(Minervini et al. 2012). T. aesticum lactic acid bacteria are
homofermentors while T. durum lactic acid bacteria are either obligate or
facultative heterofermentors.
Obligate heterofermentors are mainly found in flours rich in
fermentable carbohydrates and free amino acids suggesting a lack of
flexibility in carbon sources where a drop in readily fermentable
carbohydrates or free amino acids would lead to a rise in facultative

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heterofermentative or homofermentative lactobacilli which are less reliant on

a single carbon source. This is also the reason why the addition of bakers
yeast tends to have a negative effect on the final cell count due to the fact
that bakers yeast will offset the balance and cause a shift in the Obligate to
facultative heterofermentative and homofermentative bacteria.
There are many environmental fungus associated with bread but
Mucor plumbeus and Aspergillus niger are the two most common.
Sourdoughs show increased fungal resistance than other bread products as
Lactobacilli use linoleic acid as a substrate to produce hydroxyl fatty acids
many of which can inhibit fungal growth. Lactobacilli all oxidize linoleic acid
to coriolic acid by lipoxygenase and reductive enzymes. Lactobacilli such as,
M. plumbeus L. plantarum, L. reuteri and L. hammesii, all show large
amounts of linoleic acid consumption into fatty acids with an equivalent
amount of antifungal activity. While L. hammesii produce only one type of
monohydroxy fatty acid other lactobacilli showed the formation of a
multitude or fatty acids of varying size and structure (Black et al. 2013). The
fatty acids from L. hammesii are the most active and thus have the most
impact on fungal growth.
Fatty acids are produced largely in the first two days of growth and
fermentation while in the presence of linoleic acid, which is degraded to a
monohydroxy octadecenoic acid, as well as other enzymes, and reducing
agents from flour. Lactobacilli produce mostly 10-hydroxy-octadecenoic acid.
There is also sign that these fatty acids have a role in the cell wall

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stabilization at high temperatures

especially in L. acidophilus where high
levels of hydroxyl fatty acids are found at
high temperature growth (Black et al.
2013). Unsaturated monohydroxy fatty
acids seem to become antifungal as they inhibit fungal growth by greatly
increasing fungal cell permeability. Though, saturated and unsaturated fatty
acids have no such effect suggesting that the alcohol group plays an
important role in fungal cell permeability.
The use of di- and trihydroxy fatty acids is not common in bread
production as they produce a bitter taste in bread. Monohydroxy fatty acids
on the other hand require much higher amounts than are produced to affect
bread taste and are a viable antifungal agent in sourdough production (Black
et al. 2013).

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Refrences
Enzymes in Food Technology. R. J. Whitehurst, M. van Oort. Blackwell Publishing
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The influence of amylose and amylopectin characteristics on gelatinization and
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Andersson, A.-C. Eliasson, P. man. Volume 35, Issues 34, MarchApril 1998, Pages
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Antifirming Effects of Starch Degrading Enzymes in Bread Crumb. Hans Goesaert,
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Yeast biomass production: a new approach in glucose-limited feeding strategy. rika
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HU Meng, SONG Wei-de, SUN Can, LI Ming-ming, YANG Li-ming. Food Research and
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The Crabtree Effect: A Regulatory System in Yeast. R. H. De Deken. J . gen.
Microbiol. (1966)) 44, 149-156.
Microbial Species Diversity Interactions between Ingredients and Traditional/Typical
Italian Breads: Microbiotas of 19 Sourdoughs Used for Lactic Acid Bacterium and
Yeast. Fabio Minervini, Raffaella Di Cagno, Anna Lattanzi, Maria, Cappelle and Marco
Gobbetti, De Angelis, Livio Antonielli, Gianluigi Cardinali, Stefan. Appl. Environ.
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(Minervini et al. 2012)
Antifungal Hydroxy Fatty Acids Produced during Sourdough Fermentation: Microbial
and Enzymatic Pathways, and Antifungal Activity in Bread. Brenna A. Black,
Emanuele Zannini, Jonathan M. Curtis, Michael G. Gnzle. Appl. Environ. Microbiol.
March 2013 vol. 79 no. 6 1866-1873.
(Black et al. 2013)

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Enzymes and Bread Flavor. M. Antonia Martnez-Anaya Instituto de Agroqumica y

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Images in order from:
www.wildyeastblog.com/wp-content/uploads/2007/07/norwich-sourdough-crumb.jpg
ruralrouteramblings.files.wordpress.com/2011/08/winter-wheat01.jpg
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