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Group 6 3A-Biochemistry

21 Magsumbol
22 Mandilag
23 Marcelo
24Mendoza

Objectives
To isolate potential microorganisms as producers of

utilizable products for commercial use


To learn simple bioassay techniques for screening and
selection of microbial producer of commercial
products
To isolate and identify pure culture of antibioticproducing strain

What is molds?
Molds are fungi that grows in the form of multi cellular

filaments called hyphae.


In contrast, yeast are fungi that can adopt a single
celled growth habit.
Molds usually appear as a white fluffy fuzzy line.
The dusty texture of molds is caused by profuse
production of asexual spores.

Importance of molds
Molds play important roles in biotechnology and food

science in the production of various foods, beverages,


antibiotics, pharmaceuticals and enzymes.
Ex. The accidental discovery of the antibiotic penicillin
that is produced by the mold Penicillium
chrysogenum.

Importance of molds.
Some of the products of metabolism have great

importance.
Ex. The mold called Rhizopus produces fumaric acid
which can be used In the production of the drug
cortisone.
Other molds can also produce alcohol, citric acid and
oxalic acid.

What is antibiotics?
Antibiotics are a very important part of modern

medicine.
Antibiotics only work only against infections that are
caused by bacteria. These drugs are not effective at all
against most viral infections.
Some antibiotics are effective against only certain
types of bacteria; others can effectively fight a wide
range of bacteria.

Common
soil
molds
2 most common soil molds are Aspergillus and
Penicillium
Penicillium chrysogenum
The source for penicillin (the first antibiotic.)
Penicillin works against gram-positive bacteria, such

as Staphylococcus and Pneumococcus by disrupting


bacterial cell wall synthesis.

Penicillium
chrysogenum

The clear zone of inhibition of growth


of S. aureus with the mold P.
chrysogenum.

Common soil molds (Contd)


Aspergillus
Table 1. Zone of Inhibition (mm) Test Organisms by perpendicular
Streak Method of Fungal Isolates (According to Makut and
Owolewa)

Fungal
Isolates

C. albicans

E. coli

P.
aeruginos
a

S.
aureus

A. niger

(10 mm)

(15 mm)

(7 mm)

(16 mm)

A. flavus

(12 mm)

(8 mm)

No effect

(12 mm)

A.
fumigatus

(11 mm)

No effect

(13 mm)

(14 mm)

Aspergillus niger

Aspergillus fumigatus

Aspergillus flavus

METHODOLOGY

Weigh
Dilute
Inoculate

Incubate
Identify

Inoculate
Incubate

Examine

10g Soil Sample


Until 1:100,000
1mL in Potato Dextrose Agar by
Spread Plate Technique
72 hours at room temperature
Aspergillus
Penicillum
Isolate each mold and inoculate to
Nutrient agar vertically
Inoculate test microorganism across
making a perpendicular streak
48 hours at room temperature
Zone of inhibition

RESULTS

Botanical
A

C
B

Zone of
Inhibition

Test
organism

Gram
T.O.

5.50mm

P. aeruginosa

6.30mm

S. aureus

4.50mm

P. aeruginosa

Botanical
D
E
F

Zone of
Inhibition

Test
organism

Gram
T.O.

8.50mm

P. aeruginosa

11.80mm

P. aeruginosa

12.25mm

P. aeruginosa

FLR Laboratory Soil

Zone of
Inhibition

Test
organism

Gram
T.O.

26.25mm

P. aeruginosa

15.95mm

P. aeruginosa

16.60mm

P. aeruginosa

Sludge

Zone of
Inhibition

Test organism Gram


T.O.

23.50mm

P. aeruginosa

16.70mm

P. aeruginosa

Summary of Results
Source

Zone of
Inhibition

Test Organism

Gram Stain
T.O.

Botanical

5.50mm

P. aeruginosa

Botanical

6.30mm

S. aureus

Botanical

4.50mm

P. aeruginosa

Botanical

8.50mm

P. aeruginosa

Botanical

11.80mm

P. aeruginosa

Botanical

12.25mm

P. aeruginosa

FLR

26.25mm

P. aeruginosa

FLR

15.95mm

P. aeruginosa

FLR

16.60mm

P. aeruginosa

Sludge

23.50mm

P. aeruginosa

Sludge

16.70mm

P. aeruginosa

CONCLUSION
The group therefore conclude that molds isolated from

sludge, botanical garden, and FLR soil is a potential


antibiotic producers.
Majority of the isolated antibiotic producers came
from the Botanical Garden because of frequent
flooding.
Majority inhibits gram-negative bacteria such as
Pseudomonas aeruginosa
Some inhibits gram positive bacteria such as
Staphylococcus aureus

The most potent antibiotic producer isolated was

isolated from FLR Lab. A zone of inhibition, 26.25mm,


was observed.

References:
Penicillium chrysogenum (aka P. notatum), the natural source for the wonder
drug penicillin, the first antibiotic. Tom Volk's Fungus of the Month for
November 2003. [http://botit.botany.wisc.edu/toms_fungi/nov2003.html]
Date retrieved 6 November 2014
Makut, M. D., & Owolewa, O. A. (2011). Antibiotic-Producing Fungi Present in
the Soil Environment of Keffi Metropolis, Nasarawa State, Nigeria. Trakia
Journal of Sciences, 9(2), 33-39.