Antiobiotic Isolation From Soil

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Group 6 3A-Biochemistry21 Magsumbol

22 Mandilag23 Marcelo

24Mendoza

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ObjectivesTo isolate potential microorganisms as producers ofutilizable products for commercial use

To learn simple bioassay techniques for screening andselection of microbial producer of commercialproductsTo isolate and identify pure culture of antibiotic-

producing strain

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What is molds?Molds are fungi that grows in the form of multi cellularfilaments called hyphae.

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

production of asexual spores.

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Importance of moldsMolds play important roles in biotechnology and foodscience in the production of various foods, beverages,antibiotics, pharmaceuticals and enzymes.Ex. The accidental discovery of the antibiotic penicillinthat is produced by the mold Penicilliumchrysogenum.

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

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

oxalic acid.

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What is antibiotics? Antibiotics are a very important part of modernmedicine.

Antibiotics only work only against infections that arecaused by bacteria. These drugs are not effective at allagainst most viral infections.Some antibiotics are effective against only certain

types of bacteria; others can effectively fight a widerange of bacteria.

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Common soil molds2 most common soil molds are Aspergillus and PenicilliumPenicillium chrysogenum

The source for penicillin (the first antibiotic.)Penicillin works against gram-positive bacteria, suchas Staphylococcus and Pneumococcus by disruptingbacterial cell wall synthesis.

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The clear zone of inhibition of growthof S. aureus with the mold P.

chrysogenum .

Penicillium chrysogenum

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Common soil molds (Contd)

Aspergillus

FungalIsolates

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)

Table 1. Zone of Inhibition (mm) Test Organisms by perpendicularStreak Method of Fungal Isolates (According to Makut andOwolewa)

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Aspergillus niger Aspergillus fumigatus

Aspergillus flavus

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Weigh 10g Soil Sample

Dilute Until 1:100,000

Inoculate 1mL in Potato Dextrose Agar by

Spread Plate Technique

Incubate 72 hours at room temperature

Identify

Aspergillus Penicillum

Inoculate

Isolate each mold and inoculate toNutrient agar vertically

Inoculate test microorganism acrossmaking a perpendicular streak

Incubate 48 hours at room temperature

Examine Zone of inhibition

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RESULTS

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Botanical

Zone ofInhibition

Testorganism

GramT.O.

A 5.50mm P. aeruginosa -B 6.30mm S. aureus +

C 4.50mm P. aeruginosa -

A

BC

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Botanical

Zone ofInhibition

Testorganism

GramT.O.

D 8.50mm P. aeruginosa -

E 11.80mm P. aeruginosa -

F 12.25mm P. aeruginosa -

D

E

F

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FLR Laboratory Soil

Zone ofInhibition

Testorganism

GramT.O.

G 26.25mm P. aeruginosa -

H 15.95mm P. aeruginosa -

I 16.60mm P. aeruginosa -

GH

I

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Sludge

Zone ofInhibition Test organism GramT.O.

J 23.50mm P. aeruginosa -

K 16.70mm P. aeruginosa -

J K

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Summary of ResultsSource Zone ofInhibition

Test Organism Gram StainT.O.

A Botanical 5.50mm P. aeruginosa -

B Botanical 6.30mm S. aureus +C Botanical 4.50mm P. aeruginosa -

D Botanical 8.50mm P. aeruginosa -

E Botanical 11.80mm P. aeruginosa -

F Botanical 12.25mm P. aeruginosa -

G FLR 26.25mm P. aeruginosa -

H FLR 15.95mm P. aeruginosa -

I FLR 16.60mm P. aeruginosa -

J Sludge 23.50mm P. aeruginosa -

K Sludge 16.70mm P. aeruginosa -

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CONCLUSIONThe group therefore conclude that molds isolated fromsludge, botanical garden, and FLR soil is a potentialantibiotic producers.Majority of the isolated antibiotic producers camefrom the Botanical Garden because of frequentflooding.

Majority inhibits gram-negative bacteria such asPseudomonas aeruginosaSome inhibits gram positive bacteria such asStaphylococcus aureus

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The most potent antibiotic producer isolated wasisolated from FLR Lab. A zone of inhibition, 26.25mm, was observed.

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References:Penicillium chrysogenum (aka P. notatum), the natural source for the wonderdrug penicillin, the first antibiotic. Tom Volk's Fungus of the Month forNovember 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 inthe Soil Environment of Keffi Metropolis, Nasarawa State, Nigeria. Trakia Journal of Sciences, 9(2), 33-39.