Extraction and Purification of Biologically Active Metabolites from Rhodococcus sp. MTM3W5.2

• Main Author: Alenazi, Mohrah
• Format: Others
• Language: English
• Published: Digital Commons @ East Tennessee State University 2018
• Subjects: Antibiotics resistance; Natural product; Rhodococcus; Extraction; Purification; High-Performance Liquid Chromatography; Organic Chemistry
• Online Access: https://dc.etsu.edu/etd/3507; https://dc.etsu.edu/cgi/viewcontent.cgi?article=4958&context=etd

Rhodococcushas been recognized as a potential antibiotic producer. Recently, a strain of Rhodococcussp. MTM3W5.2 was isolated from a soil sample collected in Morristown, Tennessee and was found to produce an inhibitory compound which is active against other related species. The purpose of this research is to extract, purify and analyze the active metabolite. The compound was extracted from RM broth cultures and purified by preliminary fractionation of crude extract through a Sephadex LH-20 column. Further purification was completed using semi-preparative reversed phase column chromatography. Final purification was obtained using multiple rounds of an analytical C18 HPLC column. Based on the results achieved in the UV-Vis spectroscopy and high-resolution mass spectroscopy, the two desired compounds at a retention time of at 57 and 72 min could be polyketides with the molecular formulas C52H78O13 and C19H32O1N1/C13H34O1N1, respectively.