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STUDIES OF ANTIBACTERIAL PROPERTIES AND BIOACTIVE COMPOUNDS
Transcript of STUDIES OF ANTIBACTERIAL PROPERTIES AND BIOACTIVE COMPOUNDS
Ability to prolong life, prevent disease, preserve good health and also promote beauty.
Prevention for cancer, heart disease, lowers blood pressure, cholesterol & also prevent ulcers and kill bacteria.
Seaweeds also rich in bioactive compounds that potentially become functional ingredients for both human and animal health applications (Gupta et al., 2010)
(Cordeiro et al,2006) seaweed represents a potential source of antimicrobial substances due to their diversity of secondary metabolites with antiviral, antibacterial and antifungal activities.
STUDIES OF ANTIBACTERIAL PROPERTIES AND BIOACTIVE COMPOUNDS (Protein) FROM MARINE SEAWEED (Padina sp)
NOOR AFIFAH BT AZMI
Bachelors of Applied Sciences (Hons) Industrial Biotechnology
RESULTS & DISCUSSION
Classified based on their pigments and
colouration that exists in the plant.
Common Name: Scroll Algae, Potato Algae
Depth Collected: 3 - 40 feet
This species of seaweed
can be eaten
in some place and also been used in
Padina sp also is the only one brown seaweed that incorporated calcium.
Contains more than
72 trace minerals.
Substance or compound that has a biological activity if it has direct effects on living organism
An agent that inhibits bacterial growth or kills bacteria.
The term is often used synonymously with the term antibiotic
Pulau Besar, Mersing in Johor Bahru.
QUANTITATIVE & QUALITATIVE ANALYSIS
Accurate methods for quantifying proteins.
Measure the absorbance of the
unknown sample at a wavelength of
Seaweeds were carefully
--> removed the epiphytes
--> endoparasitic infections and undesired particles
Air dried under the shade
for 2 weeks
Gently brushed under running
seawater, rinsed with distilled
Sample ground with homogenizer
Padina sp added with distilled water and kept at 4ºC for 12 h
centrifugation at 8000rpm for 40min,
at 4 ºC.
Supernatant was collected and kept at 4ºC
Salmonella typhi Pseudomonas aeruginosa
Staphylococcus aureus Bacillus Subtilis
Centrifuge at 8000rpm for
40 mins at room temperature.
Supernatant was collected and combined with previous supernatant
Crude extract +
25% of cold (TCA)
Kept in an ice bath for 30 min.
Supernatant was discarded
8000 rpm for 40 mins at room temperature
with 10% of cold TCA &
centrifuged 8000 rpm, at 4ºC for 5 min
-> Supernatant was discarded
-> Added 5% of TCA and go for last centrifugation
1 ml of 1N NaOH
1 hour with occasionally shaking room temperature
Tubes incubated at room
temperature for 30 mins
Absorbance was determined
Six test tubes were prepared with BSA of 20,30,40,50 and 60 µl into five respectively test tubes.
2 ml of solution which consists of 48 ml 2% Na2CO3 in 0.1 N NaOH, 1ml of 1% NaK Tartrate in H2O and 1ml of 1% NaK Tartrate in H2O was added
Tubes were incubated for 10 mins
0.2 ml of dilute Folin-phenol
was added to each tube
Supervisor : Dr.Natanamurugaraj
Broth tube dilution method
24h of incubation at 37 ºC,
tubes were evaluated by
turbidity of the broth.
Method for determining the total protein
in a sample.
4 test tubes
Each tube was added with 1ml of biuret
reagent. (observed for any the
Bioactive Compound by using GC-MS Analysis & HPLC
1µl of algae extract was injected into the GC-MS with Helium carrier gas
-> flow rate of 1.50 ml/min; injector and
->column oven temperature 280ºC and 80ºC;
injection mode ‘split’ and split ratio 20:1.
-> Acetonitrile & deionised water.
->Filter first by the vacuum pump and membrane use was PTFE membrane filter. By using injection method of 20µl with flow rate of 1ml per minute and around 45 min for
each sample to be completed.
The wavelength used was 278nm-340nm.
Results & Discussion
BIOACTIVE COMPOUNDS FROM GS-MS AND HPLC
Crude protein of Padina sp
Qualitative analysis of protein already been carried
out by Biuret’s test.
Determined the lowest concentration of crude extract
against specific bacteria.
Data of Minimal inhibitory concentration (MIC) for
reaction of gram-negative E.coli.
D) Zone of Inhibition of Staphylococcus aureus
C) Zone of Inhibition Pseudomonas aeruginosa
A) Zone of Inhibition Salmonella Typhi
B) Zone of Inhibition Escherichia coli
The result shown that :
-> Padina sp clearly has the antibacterial
->Aided of HPLC and GC-MS shown more strong evidence towards the antibacterial properties
and bioactive compounds that present
E) Zone of Inhibition of Bacillus subtilis
QUALITATIVE & QUANTITATIVE
0.163 = 20.2µg approximately per 1ml
(Rollins et al., 2000) mentioned that tube dilution is the standard method for determining the levels of resistance to an antibiotic
MIC= 80µg/ml with the level of OD is 0.410