Title |
Spectral characterization of a pteridine derivative from cyanide-utilizing bacterium Bacillus subtilis - JN989651 |
Author |
S. Durairaju Nisshanthini1, Antony K. Teresa Infanta S.1, Duraisamy Senthil Raja2, Karuppannan Natarajan3, M. Palaniswamy4, and Jayaraman Angayarkanni1* |
Address |
1Department of Microbial Biotechnology, Bharathiar University, Coimbatore, Tamil Nadu, India, 2Department of Chemistry, National Tsing Hua University, Hsinchu, Taiwan, 3Department of Chemistry, Bharathiar University, Coimbatore, Tamil Nadu, India, 4Department of Microbiology, Karpagam University, Coimbatore, Tamil Nadu, India |
Bibliography |
Journal of Microbiology, 53(4),262-271, 2015,
|
DOI |
10.1007/s12275-015-4138-0
|
Key Words |
cyanide monooxygenase, Bacillus subtilis, HPLC, ESI-MS, NMR, 6-propionyl pterin |
Abstract |
Soil and water samples were collected from various regions
of SIPCOT and nearby Vanappadi Lake, Ranipet, Tamilnadu,
India. Based on their colony morphology and their stability
during subculturing, 72 bacteria were isolated, of which 14
isolates were actinomycetes. Preliminary selection was carried
out to exploit the ability of the microorganisms to utilize
sodium cyanate as nitrogen source. Those organisms
that were able to utilize cyanate were subjected to secondary
screening viz., utilization of sodium cyanide as the nitrogen
source. The oxygenolytic cleavage of cyanide is dependent
on cyanide monooxygenase which obligately requires pterin
cofactor for its activity. Based on this, the organisms capable
of utilizing sodium cyanide were tested for the presence of
pterin. Thin layer chromatography (TLC) of the cell extracts
using n-butanol: 5 N glacial acetic acid (4:1) revealed that
10 out of 12 organisms that were able to utilize cyanide had
the pterin-related blue fluorescent compound in the cell
extract. The cell extracts of these 10 organisms were subjected
to high performance thin layer chromatography (HPTLC)
for further confirmation using a pterin standard. Based on
the incubation period, cell biomass yield, peak height and
area, strain VPW3 was selected and was identified as Bacillus
subtilis. The Rf value of the cell extract was 0.73 which was
consistent with the 0.74 Rf value of the pterin standard
when scanned at 254 nm. The compound was extracted and
purified by preparative High Performance Liquid Chromatography
(HPLC). Characterization of the compound was
performed by ultraviolet spectrum, fluorescence spectrum,
Electrospray Ionization-Mass Spectrometry (ESI-MS), and
Nuclear Magnetic Resonance spectroscopy (NMR). The compound
is proposed to be 6-propionyl pterin (2-amino-6-
propionyl-3H-pteridin-4-one). |