Part III.

Making bread (application)

Part III. Making bread (application) 
Materials 
The used equipment was a basket, a plastic tub, a set of five knives, a set of planes, an
oven, different types of grill, a spatula, two sets of tablespoon and teaspoon. 
The materials of yeast bread included one half cup of whole wheat flour and one fourth
cu of white flour, one half teaspoon of yeast, one fourth teaspoon of salt, and a 90 mL of
warm water at 55oC. 
The materials of soda bread included of one-half cup of white flour, one-half of
white meal flour, third eight teaspoon of baking soda, one fourth teaspoon of salt and
one-half of butter milk. 
The 1 cup of butter milk was 1 cup of milk mixed one table spoon of vinegar. 
Method 
There are two types of yeast used to make bread: instant dried yeast and butter milk.
The method in this experiment was combined and kneaded all the ingredients. With
instant dried yeast, each group performed at least 30 minutes for incubation’s step.
With butter milk, the incubation’s time was not counted, but the butter milk should be
poured to the mixing just before baking time. All the breads were baked at 180oC
within 30 minutes.  
2. Result 
4. Making bread following two recipes 
a) Using instant dried yeast: 
Comment on the yeast bread: The product got a crispy crust, sweet taste, good smell,
and softer than the soda bread. 
 
 
b) Using baking soda: 
Comment on the soda bread: The bread tastes was soapy, salty but was tougher. The
crust was too dark with dry in texture and came denser than the yeast one.  
  
3. Discussion 
a. Protein 
- DNS assay for reducing sugar estimation
3,5-Dinitrosalicylic acid is an aromatic compound that reacts with reducing sugars to form 3-
amino-5-nitrosalicylic acid, which strongly absorbs light at 540 nm.This will result in a solution
that has red brown color whose color intensity depends on the amount of 3-amino-5-nitro
salicylic acid is formed. The color intensity of a sample then can be inferred by measuring its
absorbance, and from which the concentration of reducing sugar can be calculated throughout
the standard curve. In our group’s experiments, we noticed the red brown color in our sample
test tube, which can be a little hard to differentiated from the test tube containing blank and
control solutions. Since the standard curve calculated by the TA is excellent with the R2 = 0.987
and consistency of the absorbance over three samples, the concentration of sugar can be
inferred with confidence to be 26.6 mg/mL.
- Bradford assay for total protein quantification
The purpose of this task is to determine the concentration of unknown protein throughout
Bradford assay. The reagent (Coomassie Brilliant Blue G-250) in the Bradford assay associates
with proteins changes color from red to blue. This reaction caused the amino acids to change
blue in color, thus, the reagent-protein complex can be analyzed through the
spectrophotometer as it is within the wavelength of the visible light spectrum. There are 20
amino acids in general but Coomassie dye tends to bind with aromatic groups (phenylalanine,
tyrosine, tryptophan) and basic side chains (lysine, arginine, and histidine). CBB G exists in 3
forms with 3 colors respectively: anionic (blue), neutral (green), and cationic (red). The red form
of the dye is converted into blue color when negatively charged of the dye binds to the positive
amine groups of the protein under acidic conditions. If there is no protein to bind, the solution
will remain brown. The standard curve also is provided by the TA with a good R2 value (0.974).
According to the data and standard curve, we can calculate the concentration of unknown
sample and the result was 1.64 mg/ml.
- Enzyme activity
The enzyme activity was defined as moles of substrate converted per unit time. The recorded
results of enzyme activity was 0.54.
b. Bread 
Baking soda is a simple base and therefore needs to be combined with an acid (in our
experiment, vinegar was used) in order to become activated. The mixing of soda bread cannot
wait too much due to the rapid nature of the gas bubbles that are produced through the
reaction of acid plus sodium bicarbonate. That is the reason why the buttermilk should be
poured just before baking time.  
The product got the taste soapy, salty, bitter, and the crust is too dark, the texter is dry due to
some problems. First, our group might have added too much baking soda and salt. Secondly, our
group might have spent too much time to waiting for the dough after buttermilk was poured.
Baking soda starts to react and release its gas as soon as it comes into contact with the sour
milk. Take too long and the gas will escape before the bread is baked then lead to dry bread. 
Yeast differs from baking soda, mainly because it is a live organism and takes substantially
longer to leaven the dough. So, in the experiment, the mixing needs to incubated at least 30
mins. Unlike baking soda, yeast leavens dough through a biological process and results in
fermentation. Through fermentation, yeast can affect the taste associated with dough through
residual alcohol, making it a great option for bread. That's why there is a bit of sweetness in the
taste of the product. 
In conclusion, our product proposed that yeast is better than baking soda. 
3. Conclusion
T. reesei is an important commercial and industrial micro-organism due to its cellulase
production ability. The focus of this fermentation laboratory course was to investigate
Trichoderma sinense which was fermented at laboratory level, which began with the
preparation of cultivation medium, following by sub-culture, morphology observation and
screening for cellulase produced by the fungus. Medium preparation and subculture were
performed well using aseptic technique. However, the result was not as expected due to the
long culture time undergoing Covid 19. The sample then re-prepared for all the group by
teaching assistant and enzyme activity of cellulase was estimated 0.54 enzyme unit. In addition,
an application of yeast was performed in making bread. Type of yeasts and incubation time is
the main points that would change the taste and scent of bread.