Isolation and Characterization of a Novel Thermophilic-Organic Solvent Stable Lipase From <Emphasis Type="Italic">Acinetobacter baylyi</Emphasis> |
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Authors: | Sasithorn Uttatree Pakorn Winayanuwattikun Jittima Charoenpanich |
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Institution: | 1.Biological Science Program and Center of Excellence for Innovation in Chemistry (PERCH-CIC),Faculty of Science, Burapha University,Bangsaen,Thailand;2.Department of Biochemistry, Faculty of Science,Chulalongkorn University,Bangkok,Thailand;3.Biofuel Production by Biocatalyst Research Unit, Faculty of Science,Chulalongkorn University,Bangkok,Thailand;4.Department of Biochemistry, Faculty of Science,Burapha University,Bangsaen,Thailand;5.Environmental Science Program and Center of Excellence on Environmental Health, Toxicology and Management of Chemicals (ETM-PERDO), Faculty of Science,Burapha University,Bangsaen,Thailand |
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Abstract: | The benzene tolerant Acinetobacter baylyi isolated from marine sludge in Angsila, Thailand could constitutively secrete lipolytic enzymes. The enzyme was successfully
purified 21.89-fold to homogeneity by ammonium sulfate precipitation and gel-permeable column chromatography with a relative
molecular mass as 30 kDa. The enzyme expressed maximum activity at 60°C and pH 8.0 with p-nitrophenyl palmitate as a substrate and found to be stable in pH and temperature ranging from 6.0-9.0 to 60-80°C, respectively.
A study on solvent stability revealed that the enzyme was highly resisted to many organic solvents especially benzene and
isoamyl alcohol, but 40% inhibited by decane, hexane, acetonitrile, and short-chain alcohols. Lipase activity was completely
inhibited in the presence of Fe2+, Mn2+, EDTA, SDS, and Triton X-100 while it was suffered detrimentally by Tween 80. The activity was enhanced by phenylmethylsulfonyl
fluoride (PMSF), Na+, and Mg2+ and no significant effect was found in the presence of Ca2+ and Li+. Half of an activity was retained by Ba2+, Ag+, Hg+, Ni2+, Zn2+, and DTT. The enzyme could hydrolyze a wide range of p-nitrophenyl esters, but preferentially medium length acyl chains (C8-C12). Among natural oils and fats, the enzyme 11-folds favorably catalyzed the hydrolysis of rice bran oil, corn oil, sesame
oil, and coconut oil in comparison to palm oil. Moreover, the transesterification activity of palm oil to fatty acid methyl
esters (FAMEs) revealed 31.64 ± 1.58% after 48 h. The characteristics of novel A. baylyi lipase, as high temperature stability, organic solvent tolerance, and transesterification capacity from palm oil to FAMEs,
indicate that it could be a vigorous biocatalyzer in the prospective fields as bioenergy industry or even in organic synthesis
and pharmaceutical industry. |
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