Fungi Isolation and screening for potential Inhibitors against the two-component system and the serine/threonine protein kinase in Actinomycetes

Vun, Su Chiun (2005) Fungi Isolation and screening for potential Inhibitors against the two-component system and the serine/threonine protein kinase in Actinomycetes. Universiti Malaysia Sabah. (Unpublished)


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The aim of this research study was to isolate different strains of fungi and conduct a screening test by the fungi extract (secondary metabolites) against the twocomponent system and ser/thr protein kinase which can be applied in treating disease such as tuberculosis (TB) disease. A total 12 soil samples were collected from different sites of Danum Valley tropical rainforest, Lahad Datu under identified diterocarps trees. A total of 36 strains of fungi had been isolated by using the Potato Dextrose Agar (PDA) media (Booth, 1971; Johnston & Booth, 1983) with chloramphenicol and sodium chloride at pH 6.7 and pH 4.4. Chloramphenicol is a type of antibiotic which inhibit the growth of the unwanted bacteria. All the fungi cultures showed well and good growth on the PDA media which supplemented with rich required nutrients. The sufficient nutrients provide enough required energy and raw material source for its growth as well as its cell proliferation. The fungal cultures were then purified on the same media without chloramphenicol and also sodium chloride (NaCI). The morphology of the cultures were analyzed, observed and recorded. The morphology that was mentioned was such as the fungi's aerial mycelia, substrate mycelia and their extracellular pigment colour. Fermentation with aerobic condition was carried out after the fungi were inoculated on the PDA slant agar media and the silica gel stock for stock storage usage. The fungal colonies were inoculated onto the 10ml fermentation media inside the 125m1 conical flask. This process was carried out for 120 hours (5 days) at 28°C on the shaker incubator machine that's rotated at 220 r. p. m. Cell harvesting was carried out after the period with the 10ml of acetone for the cell lysis. The fungi extracts were then poured inside McCartney Bottle. The acetone extracts were then tested against the Two-Component System (PhoP/PhoR) in M. smegmatis H8000 and Ser/Thr Protein Kinase (AfsK/AfsR) in S. griseus H10000. A total of 4 extracts showed as potential inhibitors against the PhoP/PhoR pathway in M. smegmatis (mc1255) strain H8000. The 4 extracts with strain number H9984, H9989, H9990 and H9995 showed inhibition zone surround the paper discs on 100μM MgSO4.7H2O plate where the growth of the M. smegmatis was inhibited and not on the 1mM plate where none of the M. smegmatis growth was inhibited by the extracts for both of the screening test which been conducted. A total of 5 extracts (H9386, H9387, H9407, H9409 and H9986) that showed activity on the 1.5% D(+)Glucose plate where inhibition zone seems to be appeared surround the paper discs which been soaked with the 5 extracts above but not on the 1.5% DMannitol plate. These 5 extracts expected to inhibit either only the sporulation or the entire growth of the S. griseus strain 1-110000 where none of the microscopic observation was been carried out to determine for the potential inhibitor among the 5 extracts

Item Type: Academic Exercise
Subjects: Q Science > Q Science (General)
Department: SCHOOL > School of Science and Technology
Date Deposited: 13 May 2013 13:37
Last Modified: 13 Oct 2017 16:53

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