Isolation and purification of actinomycetes that serve as potential inhibitor of GSK3

Teh, Soo Chin (2005) Isolation and purification of actinomycetes that serve as potential inhibitor of GSK3. School of Science Technology. (Unpublished)


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The main objective of the project was to isolate and purify actinomycetes from primary lowland rainforest in Danum Valley that might serve as potential inhibitor against GSK-3p through screening system using transformed yeasts. A total of 12 soil samples have been collected. The isolation of actinomycetes was achieved through the use of humic acid-vitamin agar media. From the isolation media, a total of 31 actinomycetes were successfully purified using Oatmeal media agar. Glycerol stocks were prepared from these purified actinomycetes for storage purpose using 20% glycerol. These 31 strains from the Oatmeal media agar were then grown in 10ml mannitol-peptone-glucose liquid media separately to obtain the secondary metabolites produced by them. After duration of 120 hours in shaking incubator, 10ml acetone was added into each liquid culture to lyse the cells and release the secondary metabolites. These acetone extracts were used for screening against GSK-3 using yeast strains H10, 075 and H10, 079, with the former transformed with pKTlO-gsk3P and the latter transformed with YEP24-MCKl. The screening system is based on temperature sensitivity, in which these two transformed yeast strains are able suppress temperature sensitivity phenotype. The diameter of the halos formed around the discs at the 2 different temperatures (37°C and 25°C) determined the activity of the extract on the yeast. The formation of halos with bigger diameter at 37°C than that of 25°C indicated that the extract has potential inhibitory activity on the yeast. On the other hand, smaller halo at 37°C showed that the extract was toxic to the yeast. From a total of 115 extracts that were screened against GSK-3, only three (H11462, H11490 and H11647) of them showed potential inhibitory activity.

Item Type: Academic Exercise
Uncontrolled Keywords: actinomycetes, Danum Valley, screening system, transformed yeast, humic acid-vitamin, Oatmeal media agar, GSK-3, inhibitory activity
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: SCHOOL > School of Science and Technology
Date Deposited: 09 Jan 2014 05:59
Last Modified: 10 Oct 2017 07:57

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