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Licensed Unlicensed Requires Authentication Published by De Gruyter May 15, 2016

Pseudomonas aeruginosa strain BUP2, a novel bacterium inhabiting the rumen of Malabari goat, produces an efficient lipase

Kizhakkepowathial Nair Unni , Prakasan Priji , Sreedharan Sajith , Panichikkal Abdul Faisal and Sailas Benjamin EMAIL logo
From the journal Biologia

Abstract

Pseudomonas aeruginosa strain BUP2 (MTCC No. 5924), a novel bacterium isolated from the rumen of the Malabari goat was explored in this study for its efficiency in the production of lipase in Benjamin Unni Priji medium supplemented with 1% groundnut oil. Plackett-Burman and Box-Behnken designs were applied for optimizing the culture parameters statistically for the enhanced production of lipase; and temperature (28°C), pH (6) and incubation time (24 h) were found as significant factors for increasing the production of lipase by 11% (from 152 to 171 U/mL). Using (NH4)2SO4 fractionation and Sephadex G-100 gel filtration techniques, the lipase was purified to homogeneity (36 folds with 20% yield) with 2,392 U/mg specific activity; its apparent MW was 29 kDa, as judged by SDS-PAGE. The maximum activity (2,802 U/mL and 177% specific activity) of the purified lipase was observed with 50 mM para-nitrophenyl palmitate as substrate (at pH 8, 45°C temperature, 5.0 mM Ca2+ and 0.5% Triton X-100, after 30 min of incubation). The Km and Vmax values of the purified lipase were found as 4.75 mM and 999 μmol/min/mg, respectively; and that this may be the first report on a lipase produced by a microorganism inhabiting the rumen of a goat. Briefly, the alkalophilic and thermotolerant lipase produced by P. aeruginosa strain BUP2 with higher specific activity would find better utility in detergency; moreover, this low MW protein is a good candidate for genetic engineering toward catalytic resolution of fine chemicals.

Acknowledgements

A research grant (Order No. 1417/2014/KSCSTE) from Kerala State Council for Science, Technology & Environment, Government of Kerala, is gratefully acknowledged. The authors also declare that there exist no competing interests.

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Abbreviations
BUP

Benjamin Unni Priji

EDTA

ethylenediaminetetraacetic acid

ME

β-mercaptoethanol

pNPP

paranitrophenyl palmitate

RSM

response surface methodology

SDS

sodium dodecyl sulphate

Received: 2015-12-26
Accepted: 2016-4-14
Published Online: 2016-5-15
Published in Print: 2016-5-1

© Institute of Molecular Biology, Slovak Academy of Sciences

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