Molecular and Biochemical Characterization of Oil Degrading Bacteria
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Date
2016-07
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DEPARTMENT OF PLANT MOLECULAR BIOLOGY AND BIOTECHNOLOGY N. M. COLLEGE OF AGRICULTURE NAVSARI AGRICULTURAL UNIVERSITY Navsari
Abstract
Present investigation was carried out on “Molecular and biochemical characterization of oil degrading bacteria”. Total
six isolates from the different area were isolated. All these six
isolates showed lipid solubilization index in the range of 5.4
to 2.33. NAULM-1 showed maximum lipid solubilization
index (5.4). NAULM-4 showed lowest 2.33 lipid solubilization
index. NAULM-2, NAULM-3, NAULM-5 and NAULM-6 gave
3.3, 2.85, 3.2 and 2.43 solubilization index respectively. All
the isolates were Gram negative rod-shaped, motile and non
spore forming bacteria. On solid media all these bacterial
isolates showed raised elevated colonies with undulated
margins. Best pH for the lipase production was neutral and the
production of the same decreases with the further increase or
decrease of the pH. Temperature 35ºC, was found optimum for
lipase production by three isolates (NAULM-1, NAULM-2 and
NAULM-4 with 8.01 U/ml, 5.72 U/ml and 4.91 U/ml lipase
production at 35ºC whereas 30ºC temperature was optimum for
NAULM-3 (4.91 U/ml lipase). One per cent concentration of
the substrate was most suitable for the lipase production.
ABSTRACT
Abstract…
To supply the organic nitrogen source, casein was best
for the isolate NAULM-1, NAULM-2 and NAULM-3. Whereas
tryptone was most preferred by NAULM-4. Among the
different inorganic nitrogen sources NaNO3 was preferred by
isolates NAULM-1, NAULM-2 and NAULM-4 whereas
maximum 5.35 U/ml lipase expressions was observed in
NAULM-3 in KNO3 as inorganic nitrogen source. Overall
NAULM-1 and olive oil was the best combination of isolate
and substrate source for the efficient lipase production in
vitro. Heavy metal Cu, Pb, Cd. drastically reduced expression
of lipase in vitro. The crude enzyme worked best with high
stability at pH 7.0 and temperature 35ºC. Activities of the
enzyme was not much affected at lower temperature up to
25ºC tested by all the isolates. All the isolates were identified
using Biolog Microlog GN2 microplates and by partial
sequencing of the 16S rDNA sequencing. Universal primer
pairs, 27F and 1525R amplified ~1.5 kb fragment of partial
16S rDNA in all the isolates. On sequencing obtained 1153 bp
sequence of NAULM-1, 999 bp sequences of sequence of
NAULM-2, 1166 bp sequence of NAULM-3 and 960 bp
sequence of NAULM-4. The results of the Biolog and
Sequencing matched. On the basis of their identification the
isolate NAULM-1 was tentatively named as Pseudomonas
pseudoalcaligenes (LCB-14) NAULM-1, isolate NAULM-2
was tentatively named as Pseudomonas aeruginosa (ABG5)
NAULM-2, isolate NAULM-3 was tentatively named as
Pseudomonas aeruginosa (M2) NAULM-3 and isolate
NAULM-4 was tentatively named as Pseudomonas aeruginosa
(PS_4) NAULM-4. All these isolates belong to pseudomonas
cluster of Gamma-Proteobacteriaceae. Pseudomonas is one of
Abstract…
the widely recognized species for the production of lipase. P.
pseudoalcaligenes had been used in the past
for bioremediation and commercial lipase production by many
of the organizations. All the isolates can be used for the
commercial lipase production; however, Pseudomonas
pseudoalcaligenes (LCB-14) NAULM-1 found most preferred
and can be exploited for the purpose.
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