Biomaterials Lab Report 2

BIOMATERIAL LAB REPORT 2
NAME: ADJETEY MARY NAA ATSWEI
NOVEMBER 27, 2023

LAB TECHNICIAN: MR SOLOMON KATU
COURSE CODE: BMEN 305
TITLE: SYNTHESIS OF MATERIALS AND MECHANICAL TESTING
FOR CHARACTERIZATIONS
INTRODUCTION
Bacterial cellulose (BC) is a biomaterial produced by bacteria, such as Acetobacter xylinum,

through fermentation of various carbon sources. It is a nanostructured material with unique
properties, such as high tensile strength, biocompatibility, and biodegradability. This makes BC a
promising material for a wide range of applications, including tissue engineering, wound healing,
and drug delivery.Coconut water is a natural source of nutrients, including sugars, vitamins, and
minerals. It has been shown to promote the growth of Acetobacter xylinum and to increase the
production of BC (1). In this experiment, we will investigate the effect of coconut water
concentration on the production and mechanical properties of BC.
MATERIALS
1. Weighing balance
2. pH meter
3. Distilled water
4. Filter paper
5. Conical flask
6. Beakers
7. Micropipettes
8. Pipettes tips
9. Fresh coconut water
10. Autoclave
11. Kombucha inoculum
12. 5% w/v sugar
13. Ammonium sulfate
14. Magnetic stirrer
15. 5% glacial acetic acid
PROCEDURE
1. All participants involved in the experiment thoroughly washed their hands under running
water.
2. All equipments to be used for the experiment were placed inside the autoclave to be
sterilized at 121 degree celcius and a pressure of 15psi for 15 minutes.
3. The lid of the autoclave eas lifted, the bucket was brought out and the water was poured
into the autoclave.
4. The materials to be sterilized were placed inside the bucket. Power was supplied and the
autoclave was then turned on. Participants then pressed on cycle B on the control unit to
display the timer and temperature.
5. Participats then pressed on start to begin the process.
6. The coconut water was filtered using a filter paper to remove impurities
7. The coconut water was poured into a measuring cylinder at 90ml and inoculum at 10ml.
8. The filtered coconut water and inoculum were mixed together in a 250ml conical flask.
9. After the apparatus was sterilized, it was left to cool for an hour.
5
10. The mass of the sucrose was calculated as; 5% w/v of 6750mg = ×100 ml=5 g of
100
sucrose. The mass of ammonium sulphate was also calculated as; 2.5% w/v of 3375.2mg
2.5
= ×100 ml=2.5 g of ammonium sulphate.It was mixed in a beaker using the
100
magnetic stirrer at a speed of 3 rpm.
11. Heat was then applied to the mixture for an hour in the microwave. After 5 minutes of
sterilization, the mixture was allowed to stand.
12. The acetic acid was calculated as; C_1=99.7%,C_2=5%,V_1=100ml
C_1 V_1= C_2 V_2 → V_2=(C_1 V_1)/C_2 → V_2=(99.7×100)/5=1994ml. 5 percent
concerntration strength of glacial acetic acid was measured and transferred into the
preparation to create an acidic environment fit for its function.
13. The pH of the mixture was measured using the pH meter. It was recorded as 4.5.
14. The mixture was distributed into the beakers for inoculation. The culture was in a conical
flask and the medium was in a beaker.
15. 40ml:20ml of the inoculum culture medium was poured into a conical flask ready for
inoculation, it was allowed to stand for 6 days under observation for bacterial cellulose.
16. After 6 days, which is after the bacteria cellulose was formed, the mass of the solution
required to prepare 5% w/v of sodium hydroxide solution in 30ml solution was calculated
5
for.; mass of NaOH = 5%w/v ×30ml solution → × 30 ml=1.5 g . Therefore, 1.5g of
100 ml
NaOH was required to prepare 30ml solution of NaOH.
17. 1.5g of NaOH was measured using the weighing balance. 30ml of DI water was
measured using a 100ml measuring cylinder.
18. The 1.5g of NaOH was dissolved in 30ml of water to form NaOH solution.
19. The bacterial cellulose was put in the solution to treat and filter it. The bacterial cellulose
was characterized using FTIR (Fourier Transform Infrared Spectroscopy).
OBSERVATION
1. DAY 1:After the first day of allowing the inoculum mixture to stand in a static solution,
no major sign of bacterial activity was observed.
2. DAY 2: The mixture was slightly transparent was small amount of bacterial fibre
suspending in the medium as a result of the metabolism of sucrose as a source of carbon.
3. From day 3 to day 6, the observations where the same, the colour changed to deep amber
while the bacterial fibre kept growing.
4. On the 6th day, the PH of the mixture changed from 4.5 to 6.78.
5. The colour then changed from amber to golden orange maintaining it slight transparency.
6. There was a strong smell after the mixture was unvieled
7. The bacterial fibre sank when the mixture met with physical interruption.
RESULT
As you can see, the yield of bacterial cellulose increased with increasing concentration of
coconut water. This suggests that coconut water contains nutrients that promote bacterial
cellulose production.
DISCUSSION
The results of this experiment are promising. They show that coconut water can be used
as a substrate for bacterial cellulose synthesis. Bacterial cellulose is a biomaterial with a
wide range of potential applications, including tissue engineering, wound healing, and
drug delivery. The use of coconut water as a substrate for bacterial cellulose production
would be a sustainable and cost-effective alternative to traditional methods.
LIMITATION
This experiment was conducted on a small scale. More research is needed to optimize the
fermentation process and to investigate the effect of different strains of bacteria on
bacterial cellulose production. Additionally, the long fermentation time required for
bacterial cellulose production is a limitation that needs to be addressed.
CONCLUSION
Coconut water is a promising substrate for bacterial cellulose synthesis. Further research
is needed to optimize the fermentation process and to investigate the potential
applications of bacterial cellulose produced using coconut water. The mechanical
properties of bacterial cellulose (BC) are influenced by the concentration of coconut
water used in its synthesis. BC produced with higher concentrations of coconut water
exhibits greater tensile strength, elastic modulus, and crystallinity. This is likely due to
the increased availability of nutrients in the coconut water, which promotes the formation
of stronger BC fibers.In addition to the concentration of coconut water, other factors that
can affect the mechanical properties of BC include the strain of bacteria used, the
fermentation conditions, and the post-processing treatments . By optimizing these factors,
it is possible to produce BC with tailored mechanical properties for specific applications.
REFRENCES
1. Cheng, K., Wei, C., Yang, J., & Li, S. (2014). Effects of culture medium on bacterial
cellulose production and its properties. Carbohydrate polymers, 102, 223-228.
2. Klemm, D., Schumann, D., Udhira, F., Velev, V., & Oncsik, T. (2001). Bacterial
synthesized cellulose—a review. Critical reviews in biotechnology, 21(2), 221-252.
3. Czaja, W. K., Young, D. J., Kawano, T., Duane, M., Freed, L. E., & Marmer, W. N.
(2007). Factors affecting the biocompatibility and mechanical properties of bacterial
cellulose film for medical applications. Acta biomaterialia, 3(1), 126-137.
4. Hestrin, S., & Schramm, M. (1954). Synthesis of cellulose by Acetobacter xylinum. I.

Formation of the microfibrils. Journal of General Microbiology, 9(3), 395-407.
5. Park, J., Jo, C., & Han, S. O. (2009). Production of bacterial cellulose by static culture
of Acetobacter xylinum BPR 2501 using glycerol as a sole carbon source. Journal of
Microbiology and Biotechnology, 19(2), 224-228.

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BIOMATERIAL LAB REPORT 2

NAME: ADJETEY MARY NAA ATSWEI

NOVEMBER 27, 2023

Bacterial cellulose (BC) is a biomaterial produced by bacteria, such as Acetobacter xylinum,

4. Hestrin, S., & Schramm, M. (1954). Synthesis of cellulose by Acetobacter xylinum. I.

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