Isolation and purification of actinomycetes for screening of secondary metabolites against MAPK kinase and MAPK phosphatase in eukaryotic signal transduction pathway

Celistha Fay J. Ferdinand Deosing, (2005) Isolation and purification of actinomycetes for screening of secondary metabolites against MAPK kinase and MAPK phosphatase in eukaryotic signal transduction pathway. Universiti Malaysia Sabah. (Unpublished)

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Abstract

A total of 16 soil samples were collected from underneath selected and identified trees in Newbery Plot of lowland dipterocarp rainforest at Danwn Valley, Lahad Datu, Sabah. The collection of soil was done around the base of dipterocarps as this area has the least tendency for cross-contamination and abundant with humus which supplies nutrients. The soil pH was measured in 1:2.5 soil-water suspension. The pH determined for the soil samples was pH 4.0 (acidic). Soil samples were first mixed, suspended in sterile dH20 and shaken on vortex mixer. All treated samples were serially diluted up to 10-3 and 0.1 ml of aliquot was spread over the surface of humic acid-vitamin B agar (HV). Plates were incubated at 28°C for growth of colonies within 7 to 14 days. Oatmeal agar (OA) was used to obtain single pure colonies. The pure cultures sporulated well on these OA media. There were 47 strains of pure actinomycetes isolated, purified and characterized by presence of aerial and substrate mycelia, extracellular pigmentation and sporulation. The colour definition for the physical and morphological characteristics was standardized according to Sissons Paint Catalogue. These 47 pure actinomycetes were grown aerobically in liquid culture for constant fermentation at 28°C for 5 days and thereafter extracted with acetone. The 47 acetone extracts were screened for microbial inhibitors against MAPK kinase (MKK1) and MAPK phosphatase (MSG5) in budding yeast (Saccharomyces cerevisiae) systems. Besides of screening those 47 extracts, a total of 101 extracts owned by other coursemates were also screened. As a result, overall there would be ]48 extracts that were screened, with a total of only 132 out of the 148 extracts tested against MKKl, while against MSG5 all of the 148 extracts were tested. Out of the 148 extracts tested, only 7 extracts showed activity (toxicity). All of the 7 extracts (H1161O, H1 1612, HI1635, HII640, H11813, H11428 & H11588) showed toxicity against MSG5, in which 2 extracts (H1 1610 & HI 1640) out of the 7 extracts that showed toxicity against MSG5, did not show any activity (nontoxic) against MKKI. This indicates that only 5 extracts (HI1612, H1 1635, H11813, H11428 & H11588) out of the 7 extracts showed toxic activity against MKKl. In this research, there was no discovery of potential inhibitors against MKKI and MSG5.

Item Type: Academic Exercise
Uncontrolled Keywords: soil sampling, actinomycetes, yeast strain, sterilization
Subjects: T Technology > TD Environmental technology. Sanitary engineering
Divisions: SCHOOL > School of Science and Technology
Depositing User: MDM SITI AZIZAH IDRIS
Date Deposited: 03 Sep 2013 03:19
Last Modified: 23 Oct 2017 07:58
URI: http://eprints.ums.edu.my/id/eprint/6853

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