Purification, characterization and thermal inactivation kinetics of a non-regioselective thermostable lipase from a genotypically identified extremophilic Bacillus subtilis NS 8.

Thermostable lipase produced by a genotypically identified extremophilic Bacillus subtilis NS 8 was purified 500-fold to homogeneity with a recovery of 16% by ultrafiltration, DEAE-Toyopearl 650M and Sephadex G-75 column. The purified enzyme showed a prominent single band with a molecular weight of...

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Main Authors: Olusesan, Akanbi Taiwo, Kamaruzaman, Azura Liana, Forghani, Bita, Abu Bakar, Fatimah, Mohammed, Abdulkarim Sabo, Radu, Son, Abd. Manap, Mohd. Yazid, Saari, Nazamid
Format: Article
Language:English
Published: Elsevier 2011
Online Access:http://psasir.upm.edu.my/id/eprint/24096/
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author Olusesan, Akanbi Taiwo
Kamaruzaman, Azura Liana
Forghani, Bita
Abu Bakar, Fatimah
Mohammed, Abdulkarim Sabo
Radu, Son
Abd. Manap, Mohd. Yazid
Saari, Nazamid
author_facet Olusesan, Akanbi Taiwo
Kamaruzaman, Azura Liana
Forghani, Bita
Abu Bakar, Fatimah
Mohammed, Abdulkarim Sabo
Radu, Son
Abd. Manap, Mohd. Yazid
Saari, Nazamid
author_sort Olusesan, Akanbi Taiwo
building UPM Institutional Repository
collection Online Access
description Thermostable lipase produced by a genotypically identified extremophilic Bacillus subtilis NS 8 was purified 500-fold to homogeneity with a recovery of 16% by ultrafiltration, DEAE-Toyopearl 650M and Sephadex G-75 column. The purified enzyme showed a prominent single band with a molecular weight of 45 kDa. The optimum pH and temperature for activity of lipase were 7.0 and 60°C, respectively. The enzyme was stable in the pH range between 7.0 and 9.0 and temperature range between 40 and 70°C. It showed high stability with half-lives of 273.38 min at 60°C, 51.04 min at 70°C and 41.58 min at 80°C. The D-values at 60, 70 and 80°C were 788.70, 169.59 and 138.15 min, respectively. The enzyme's enthalpy, entropy and Gibb's free energy were in the range of 70.07–70.40 kJ mol−1, −83.58 to −77.32 kJ mol−1 K−1 and 95.60–98.96 kJ mol−1, respectively. Lipase activity was slightly enhanced when treated with Mg2+ but there was no significant enhancement or inhibition of the activity with Ca2+. However, other metal ions markedly inhibited its activity. Of all the natural vegetable oils tested, it had slightly higher hydrolytic activity on soybean oil compared to other oils. On TLC plate, the enzyme showed non-regioselective activity for triolein hydrolysis.
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spelling upm-240962014-02-24T05:02:42Z http://psasir.upm.edu.my/id/eprint/24096/ Purification, characterization and thermal inactivation kinetics of a non-regioselective thermostable lipase from a genotypically identified extremophilic Bacillus subtilis NS 8. Olusesan, Akanbi Taiwo Kamaruzaman, Azura Liana Forghani, Bita Abu Bakar, Fatimah Mohammed, Abdulkarim Sabo Radu, Son Abd. Manap, Mohd. Yazid Saari, Nazamid Thermostable lipase produced by a genotypically identified extremophilic Bacillus subtilis NS 8 was purified 500-fold to homogeneity with a recovery of 16% by ultrafiltration, DEAE-Toyopearl 650M and Sephadex G-75 column. The purified enzyme showed a prominent single band with a molecular weight of 45 kDa. The optimum pH and temperature for activity of lipase were 7.0 and 60°C, respectively. The enzyme was stable in the pH range between 7.0 and 9.0 and temperature range between 40 and 70°C. It showed high stability with half-lives of 273.38 min at 60°C, 51.04 min at 70°C and 41.58 min at 80°C. The D-values at 60, 70 and 80°C were 788.70, 169.59 and 138.15 min, respectively. The enzyme's enthalpy, entropy and Gibb's free energy were in the range of 70.07–70.40 kJ mol−1, −83.58 to −77.32 kJ mol−1 K−1 and 95.60–98.96 kJ mol−1, respectively. Lipase activity was slightly enhanced when treated with Mg2+ but there was no significant enhancement or inhibition of the activity with Ca2+. However, other metal ions markedly inhibited its activity. Of all the natural vegetable oils tested, it had slightly higher hydrolytic activity on soybean oil compared to other oils. On TLC plate, the enzyme showed non-regioselective activity for triolein hydrolysis. Elsevier 2011 Article PeerReviewed Olusesan, Akanbi Taiwo and Kamaruzaman, Azura Liana and Forghani, Bita and Abu Bakar, Fatimah and Mohammed, Abdulkarim Sabo and Radu, Son and Abd. Manap, Mohd. Yazid and Saari, Nazamid (2011) Purification, characterization and thermal inactivation kinetics of a non-regioselective thermostable lipase from a genotypically identified extremophilic Bacillus subtilis NS 8. New Biotechnology, 28 (6). pp. 738-745. ISSN 1871-6784 10.1016/j.nbt.2011.01.002 English
spellingShingle Olusesan, Akanbi Taiwo
Kamaruzaman, Azura Liana
Forghani, Bita
Abu Bakar, Fatimah
Mohammed, Abdulkarim Sabo
Radu, Son
Abd. Manap, Mohd. Yazid
Saari, Nazamid
Purification, characterization and thermal inactivation kinetics of a non-regioselective thermostable lipase from a genotypically identified extremophilic Bacillus subtilis NS 8.
title Purification, characterization and thermal inactivation kinetics of a non-regioselective thermostable lipase from a genotypically identified extremophilic Bacillus subtilis NS 8.
title_full Purification, characterization and thermal inactivation kinetics of a non-regioselective thermostable lipase from a genotypically identified extremophilic Bacillus subtilis NS 8.
title_fullStr Purification, characterization and thermal inactivation kinetics of a non-regioselective thermostable lipase from a genotypically identified extremophilic Bacillus subtilis NS 8.
title_full_unstemmed Purification, characterization and thermal inactivation kinetics of a non-regioselective thermostable lipase from a genotypically identified extremophilic Bacillus subtilis NS 8.
title_short Purification, characterization and thermal inactivation kinetics of a non-regioselective thermostable lipase from a genotypically identified extremophilic Bacillus subtilis NS 8.
title_sort purification, characterization and thermal inactivation kinetics of a non-regioselective thermostable lipase from a genotypically identified extremophilic bacillus subtilis ns 8.
url http://psasir.upm.edu.my/id/eprint/24096/
http://psasir.upm.edu.my/id/eprint/24096/