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S. Durairaju Nisshanthini,Antony K. Teresa Infanta S.,Duraisamy Senthil Raja,Karuppannan Natarajan,M. Palaniswamy,Jayaraman Angayarkanni 한국미생물학회 2015 The journal of microbiology Vol.53 No.4
Soil and water samples were collected from various regionsof SIPCOT and nearby Vanappadi Lake, Ranipet, Tamilnadu,India. Based on their colony morphology and their stabilityduring subculturing, 72 bacteria were isolated, of which 14isolates were actinomycetes. Preliminary selection was carriedout to exploit the ability of the microorganisms to utilizesodium cyanate as nitrogen source. Those organismsthat were able to utilize cyanate were subjected to secondaryscreening viz., utilization of sodium cyanide as the nitrogensource. The oxygenolytic cleavage of cyanide is dependenton cyanide monooxygenase which obligately requires pterincofactor for its activity. Based on this, the organisms capableof utilizing sodium cyanide were tested for the presence ofpterin. Thin layer chromatography (TLC) of the cell extractsusing n-butanol: 5 N glacial acetic acid (4:1) revealed that10 out of 12 organisms that were able to utilize cyanide hadthe pterin-related blue fluorescent compound in the cellextract. The cell extracts of these 10 organisms were subjectedto high performance thin layer chromatography (HPTLC)for further confirmation using a pterin standard. Based onthe incubation period, cell biomass yield, peak height andarea, strain VPW3 was selected and was identified as Bacillussubtilis. The Rf value of the cell extract was 0.73 which wasconsistent with the 0.74 Rf value of the pterin standardwhen scanned at 254 nm. The compound was extracted andpurified by preparative High Performance Liquid Chromatography(HPLC). Characterization of the compound wasperformed by ultraviolet spectrum, fluorescence spectrum,Electrospray Ionization-Mass Spectrometry (ESI-MS), andNuclear Magnetic Resonance spectroscopy (NMR). The compoundis proposed to be 6-propionyl pterin (2-amino-6-propionyl-3H-pteridin-4-one).