Isolation, Production and Characterization of Thermostable Lipase from Bacillus Sp. Strain L2

A thermostable lipolytic endospore forming Bacillus sp. strain L2 was isolated from a hot spring (91oC) in Slim River, Perak, Malaysia. Nutritional studies showed that casamino acid was the best nitrogen source while trehalose and Tween 60 were the best carbon source and substrate respectively, f...

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Main Author:	Mohammed Mukred, Abdul Daim
Format:	Thesis
Language:	English English
Published:	2004
Subjects:	Lipase - isolation & purification Biotechnology Proteolytic enzymes
Online Access:	http://psasir.upm.edu.my/id/eprint/16/1/1000548940_t_IB_2005_13.pdf
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id	my-upm-ir.16
record_format	uketd_dc
spelling	my-upm-ir.162013-05-27T06:45:02Z Isolation, Production and Characterization of Thermostable Lipase from Bacillus Sp. Strain L2 2004-12 Mohammed Mukred, Abdul Daim A thermostable lipolytic endospore forming Bacillus sp. strain L2 was isolated from a hot spring (91oC) in Slim River, Perak, Malaysia. Nutritional studies showed that casamino acid was the best nitrogen source while trehalose and Tween 60 were the best carbon source and substrate respectively, for lipase production. The enzyme production was promoted in the presence of metal ions such as Ca2+, Cu2+, Fe2+, Co2+, Zn2+, and Sr2+. The optimum lipase production was obtained in the presence of CaCl2. Physiological study indicate that, this strain showed optimal lipase production after 28 h of incubation at 70 oC with 1% starting inoculum. The enzyme was purified to homogeneity by ammonium sulphate precipitation and affinity column chromatography (Heparin), with 3 fold increase in specific activity and 48.9 % recovery. The purified enzyme produced a single band on SDS-PAGE. Its molecular mass was estimated to be 43 KDa. The optimum pH and temperature for L2 lipase were determined to be 7.0 and 80 oC, respectively. The III lipase was very stable for 2 h at 80 oC. The stability at higher temperature was shown to be calcium dependent. The lipase activity was enhanced by Fe2+and Ca2+ while Cu2+ inhibited it. The enzyme activity was strongly inhibited by PMSF and completely inhibited by pCMB. Substrates such as olive oil and sesame oil enhanced the lipase activity. Lipase - isolation & purification Biotechnology Proteolytic enzymes 2004-12 Thesis http://psasir.upm.edu.my/id/eprint/16/ http://psasir.upm.edu.my/id/eprint/16/1/1000548940_t_IB_2005_13.pdf application/pdf en public masters Universiti Putra Malaysia Lipase - isolation & purification Biotechnology Proteolytic enzymes Faculty of Biotechnology and Biomolecular Sciences English
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English English
topic	Lipase - isolation & purification Biotechnology Proteolytic enzymes
spellingShingle	Lipase - isolation & purification Biotechnology Proteolytic enzymes Mohammed Mukred, Abdul Daim Isolation, Production and Characterization of Thermostable Lipase from Bacillus Sp. Strain L2
description	A thermostable lipolytic endospore forming Bacillus sp. strain L2 was isolated from a hot spring (91oC) in Slim River, Perak, Malaysia. Nutritional studies showed that casamino acid was the best nitrogen source while trehalose and Tween 60 were the best carbon source and substrate respectively, for lipase production. The enzyme production was promoted in the presence of metal ions such as Ca2+, Cu2+, Fe2+, Co2+, Zn2+, and Sr2+. The optimum lipase production was obtained in the presence of CaCl2. Physiological study indicate that, this strain showed optimal lipase production after 28 h of incubation at 70 oC with 1% starting inoculum. The enzyme was purified to homogeneity by ammonium sulphate precipitation and affinity column chromatography (Heparin), with 3 fold increase in specific activity and 48.9 % recovery. The purified enzyme produced a single band on SDS-PAGE. Its molecular mass was estimated to be 43 KDa. The optimum pH and temperature for L2 lipase were determined to be 7.0 and 80 oC, respectively. The III lipase was very stable for 2 h at 80 oC. The stability at higher temperature was shown to be calcium dependent. The lipase activity was enhanced by Fe2+and Ca2+ while Cu2+ inhibited it. The enzyme activity was strongly inhibited by PMSF and completely inhibited by pCMB. Substrates such as olive oil and sesame oil enhanced the lipase activity.
format	Thesis
qualification_level	Master's degree
author	Mohammed Mukred, Abdul Daim
author_facet	Mohammed Mukred, Abdul Daim
author_sort	Mohammed Mukred, Abdul Daim
title	Isolation, Production and Characterization of Thermostable Lipase from Bacillus Sp. Strain L2
title_short	Isolation, Production and Characterization of Thermostable Lipase from Bacillus Sp. Strain L2
title_full	Isolation, Production and Characterization of Thermostable Lipase from Bacillus Sp. Strain L2
title_fullStr	Isolation, Production and Characterization of Thermostable Lipase from Bacillus Sp. Strain L2
title_full_unstemmed	Isolation, Production and Characterization of Thermostable Lipase from Bacillus Sp. Strain L2
title_sort	isolation, production and characterization of thermostable lipase from bacillus sp. strain l2
granting_institution	Universiti Putra Malaysia
granting_department	Faculty of Biotechnology and Biomolecular Sciences
publishDate	2004
url	http://psasir.upm.edu.my/id/eprint/16/1/1000548940_t_IB_2005_13.pdf
_version_	1747810148613619712

Isolation, Production and Characterization of Thermostable Lipase from Bacillus Sp. Strain L2

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