Renz Andrew L.

de Lira 10-Electron

TITLE: Analysis of the Antimicrobial Activity of Lycopene (C10 H26 ) Isolated from Solanum
lycopersicum (Tomato) Against Bacillus subtilis

Introduction

I. Research Topic

Lycopene is an antioxidant that is present in red and pink-colored fruits and vegetables it has
been shown to have heart, blood pressure, prostate, osteoporosis, skin and other benefits in both
natural and synthetic forms. One of the best sources of Lycopene is Solanum lycopersicum more
commonly known as tomato. Because of its health benefits, Lycopene is speculated to house
antimicrobial properties.

This study attempts to successfully isolate Lycopene from tomato through conventional
solvent extraction and anti-solvent precipitation. After the isolation, gram-positive bacteria, Bacillus
subtilis will then be cultured in a nutrient agar. The bacteria will determine whether Lycopene exhibits
antimicrobial property or not.

Proceeding into the experiment proper, a preferable amount of 50 mg isolated Lycopene will
be included in the testing. The agar medium containing B. subtilis will then be poured into a Petri
dish. Immediately, 0.1 mL of the isolated Lycopene will be added onto the cups. Finally, it will be kept
for diffusion in a freezer for 2 hours. It will then be removed and placed in an incubator at 37˚C for
48 hours. After incubation, the zone of inhibition will be measured in millimeters and will be recorded
for the analysis of the antimicrobial activity.

II. Significance of the Problem

This research aims to analyze the possible antimicrobial property of Lycopene extracted from
tomatoes against B. subtilis. The results of the study may serve as a basis for a possible engineering
of an antibacterial drug based on Lycopene. Synthesis of an herbal alternative drug can help to lessen
side effects administered by usual antibacterial drugs. In addition, it can also make antibacterial drugs
more affordable and more efficient. An efficient and affordable alternative drug is much well-needed
by a large portion of the community.

III. Statement of the Problem

 Does Lycopene, an antioxidant isolated from Solanum lycopersicum (tomato), exhibit

antimicrobial activity against Bacillus subtilis?
o Can Lycopene be successfully isolated from S. lycopersicum through
conventional solvent extraction and anti-solvent precipitation?
o How effective is Lycopene against B. subtilis, by measuring the zone of
inhibition in millimeters?
o Is the antimicrobial property of Lycopene significant enough in order for it to
be available for commercial use?

Brief Literature Survey

Lycopene is an open-chain carotenoid found in several fruits and vegetables, especially S.

lycopersicum (tomatoes). It may accumulate in high concentrations in several tissues reacting with
hydroxyl radicals to give a stabilized, highly delocalized species. (Avendano & Menendez, 2015).

Lycopene can be isolated from S. lycopersicum through various methods. The method that
will be used in this study is by using conventional solvent extraction and anti-solvent precipitation.
This particular method has shown to be more effective than the previous reported methods (Roh et
al., 2013). However, in another study, other methods were used to extract Lycopene, and the study
showed that extracted lycopene by different methods showed antimicrobial activity against P.
aeruginosa (Maitra & Sangeeta, 2016).

Lycopene possesses antibacterial and antifungal properties. From a present investigation it

was concluded that the lycopene which was extracted from tomato was effective against oral
pathogens like Staphylococcus aureus and Candida albicans. Candida albicans is a diploid fungus that
can cause opportunistic oral and genital infections in humans. (Sung et al., 2007). In a previous
study, Lycopene was shown to possess antimicrobial property by showing effective inhibition against
the bacteria. Therefore, Lycopene can be considered to be the promising source of antimicrobial
compounds (Dhanawade & Sakhave, 2014). Other than showing antimicrobial property, researchers
also found out that Lycopene has a strong inhibitory effect on C. trachomatis and C. pneumoniae
infections in alveolar macrophages. This finding was documented in our studies by both the
immunofluorescence analysis and electron microscopy. It has to be noted that degree of lycopene
inhibitory of both chlamydia growth was overwhelming and reached over 90% according to the
immunofluorescence analysis (Zingagirova et al., 2017).

Feasibility

The chemicals needed for the isolation of Lycopene from S. lycopersicum are the following
solvents: hexane, ethyl acetate, and ethanol. These solvents can be found in the PSHS-EVC
laboratories. Methanol is also needed as an anti-solvent, which is also readily available in the school.
The bacteria, B. subtilis, can be purchased online. The incubator and the freezer needed for
measuring the zone of inhibition is also available at the laboratory. The entire experiment can be in
about a week at most.

Limitations

In the Lycopene extraction, the original method that I planned to use was by using TLC test,
UV Spectroscopy, and IR analysis, which is unfortunately not as feasible as the method that I am now
currently using.

For the bacteria sample, I wanted to include more species of bacteria, and even fungi in the
study to diversify the results, and to know which bacteria/fungi will be more susceptible to damage.
However, since there is an unavailability of materials if I were to continue with that path, I decided to
pick just one of the bacteria, which is B. subtilis.

References

Avendano, C., & Menendez, J.C. (2015). Cancer Chemoprevention. Retrieved from
https://www.sciencedirect.com/topics/chemistry/lycopene

Dhanawade, S., & Sakhave, A. (2014). Isolation of Lycopene from Tomato and Study of Its
Antimicrobial Activity. Retrieved from https://www.ijsr.net › archive

Maitra, J., & Sangeeta. (2016). Antibacterial Property of Lycopene Extracted from Lycopersicon
esculentum against Pseudomonias aeruginosa. Retrieved from
http://www.ejbps.com/admin/download/article/dm9sdW1lXzNfc2VwdGVtYmVyX2lzc3VlXzkvMTQ3MjYz
NTU1NC5wZGY=

Roh M., Jeon M., Moon J., Moon W., Park S., & Choi J. (2013). A Simple Method for the Isolation of
Lycopene from Lycopersicon esculentum. Retrieved from Isolationoflycopene-Bot.Sci.912187-192.pdf

Sung WS, Lee IS, Lee DG. (2007). Damage to the cytoplasmic membrane and cell death caused by
lycopene in Candida albicans. Retrieved from
http://www.phytochemicals.info/phytochemicals/lycopene/antibacterial-antifungal.php
Zingagirova, N., Morgunova, E., Fedina, E., Shevyagina, N., Borovaya, T., Zhukhovitsky, V., Kyle, N.,
& Petyaev, I.(2017). Lycopene Inhibits Propagation of Chlamydia Infection. Retrieved from
http://www.hindawi.com/journals/scientifica/2017/1478625/