Fay Acid Methyl Ester (FAME) Analysis of Moderately Thermophilic Bacteria
Isolated from the Coramandal Coast, Chennai, Tamilnadu
Sharmili AS
1
and Ramasamy P
2
1
Department of Biotechnology, Stella Maris College (Autonomous), Chennai 600 086, India
2
Research and Development Wing, Sree Balaji Medical College and Hospital, BIHER (Bharath University), Chennai, India
Corresponding author: Sharmili AS, Department of Biotechnology, Stella Maris College (Autonomous), Chennai 600 086, India; Email:
arsharmilis@gmail.com
Received date: October 28, 2016; Accepted date: December 28, 2016; Published date: December 30, 2016
Copyright: © 2016 Sharmili AS, et al. This is an open-access arcle distributed under the terms of the Creave Commons Aribuon License, which
permits unrestricted use, distribuon, and reproducon in any medium, provided the original author and source are credited.
Citaon: Sharmili AS, Ramasamy P. Fay Acid Methyl Ester (FAME) Analysis of Moderately Thermophilic Bacteria Isolated from the Coramandal
Coast, Chennai, Tamilnadu.
Abstract
A total of 18 out of 44 moderately thermophlic bacteria
isolated from water samples of the Coramandal Coast,
Chennai, Tamilnadu, were analyzed for Fay Acid Methyl
Ester (FAME). The present study showed that the
predominant fay acid was 15:0i followed by 15:0a, which
were characterisc of Bacillus. The results suggest that the
order of decreasing abundance of terminally branched fay
acids is as follows: iso even-numbered acids namely 16:0i,
14:0i, iso odd-numbered acids, 15:0i, 17:0i, 13:0i, 19:0i and
anteiso acids, 15:0a, 17:0a, 11:0a, 19:0a. Straight chain
saturated fay acids idenfied were 10:0, 12:0, 14:0, 16:0,
18:0. The remaining fay acids components were present in
negligible quanes. The isolates were also idenfied and
classified using the comparison with the TSBA database as
B. cereus-GC subgroup A (isolates ASR 29CIII, ASR 37CIII), B.
sublis (isolates ASR 6SII, ASR 21SI, ASR 25SV), B.
laevolaccus (isolates ASR 4LIII, ASR 7SIII, ASR 11LVI, ASR
42SVII), B. alcalophilus (isolates ASR 14PI, ASR 18PIV), B.
pumilus subgroup A (isolate- ASR 41PVIII), Staphylococcus
schleiferi (isolates- ASR 2LII, ASR 5LIV), S. gallinarum-GC
subgroup A (isolates ASR 13SIV, ASR 33SVI), Kurthia sibirica
(isolate ASR 15PII), Geobacillus stearothermophilus-GC
subgroup (isolate ASR 8LV).
Keywords: Moderately thermophilic bacteria; Bacillus;
FAME; Fay acids; 15:0i; 15:0a; 17:0a
Introducon
Microbial biomarkers are chemical components of
microorganisms which can be analyzed in a given sample and
interpreted (both quantavely and qualitavely) in terms of in
situ microbial biomass. The most useful biomarkers are
membrane lipids and their related fay acids as they are
essenal components of every living cell and have great
structural diversity and high biological specificity [1]. Two
common approaches are analysis of microbial lipids: (i) polar
phospholipid fay acid (PLFA) analysis and (ii) total fay acid
methyl ester (total FAME) [2]. PLFA assay provides informaon
leading to idenficaon and quanficaon of viable bacterial
biomass [3]. Total FAME analysis, on the other hand, includes all
saponifiable lipids present in the sample (including PLFAs).
Polyunsaturated fay acids and long chain fay acids beyond 18
carbons are absent in Prokaryotes. Saturated or
monounsaturated fay acids of length 10 to 18 carbon are
present in Eukaroytes [4,5].
Bacterial fay acids are highly conserved due to their role in
cell structure and funcon and are the major constuents of the
lipid bilayer of bacterial membranes and lipopolysaccharides.
They have been used extensively for taxonomic and
idenficaon purposes. Whole cellular FAME content is a
bacterial profile and is a direct and stable expression of the
cellular genome. The cellular fay acid paern is a phenotypic
character that is not affected by mutaons, acquision or loss of
plasmids. The use of fay acid analysis by gas chromatography
for the idenficaon of bacteria is rapid, efficient, reproducible
and used for the idenficaon of both clinical and
environmental isolates [6-10]. Fay acids are mainly located in
the cytoplasmic membrane as constuents of phospholipids and
as lipopolysaccharides in the outer membrane of Gram-negave
bacteria as well as lipoteichoic acids in Gram-posive bacteria.
The importance of FAME analysis for the idenficaon of
bacteria is based on the large structural differences within these
molecules viz., (i) variaon in length (8 to 20 C-atoms), (ii)
presence of saturated and monounsaturated fay acids, (iii)
occurrence of branched fay acids (iso and anteiso fay acids or
methylated within the molecule), (iv) occurrence of
cyclopropane fay acids (17:0c, 19:0c), (v) occurrence of
hydroxy-fay acids with an OH-group at posion two or three of
the molecule. For classificaon or idenficaon of bacteria the
presence of disnct fay acids and their relave amount is
analyzed and compared with the fay acid profiles of reference
strains. The characterisc feature of a phylogenec group of
bacteria can be the dominang presence of a single fay acid or
a specific fay acid paern. As the fay acid composion of
bacteria is dependent on the growth phase, temperature and
growth medium, standardizaon of these condions is
important. FAME assay is a powerful tool in the study of
Research Article
iMedPub Journals
http://www.imedpub.com/
European Journal of Experimental Biology
ISSN 2248-9215
Vol.6 No.6:5
2016
© Under License of Creative Commons Attribution 3.0 License | This article is available from: http://www.imedpub.com/european-journal-of-experimental-
biology/
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