Purification and characterisation of a thermostable alkaline lipase from a new thermophilic Bacillus sp. RSJ-1

Sharma, R and Soni, S.K and Vohra, R M and Gupta, L.K. and Gupta, J.K. (2002) Purification and characterisation of a thermostable alkaline lipase from a new thermophilic Bacillus sp. RSJ-1. Process Biochemistry, 37 (10). pp. 1075-1084. ISSN 13595113

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Official URL: http://dx.doi.org/10.1016/S0032-9592(01)00316-8

Abstract

An extracellular alkaline lipase from a new thermophilic Bacillus sp. RSJ-1 was purified to homogeneity by ultrafiltration, followed by ammonium sulfate precipitation, dialysis, Q-Sepharose ion exchange chromatography and Sephacryl S-200 SF gel filtration chromatography. This purification protocol resulted in a 201-fold purification of lipase with 19.7% final yield and the relative molecular weight of the enzyme was determined to be 37 kDa by SDS-PAGE. The kinetic characterisation of the purified enzyme exhibited maximum activity at 50 °C and pH 8.0–9.0. It was stable at 50 °C for 60 min and retained >90% of its original activity for 120 min. The half lives at 55, 60, 65, 70 and 75 °C were 240, 150, 90, 45 and 30 min, respectively. The enzyme was also highly stable in a pH range of 8.0–9.0 for 120 min. The enzyme activity was promoted in the presence of Ca2+, Na+, Mg2+ and Ba2+ and was strongly inhibited by Cs+, K+, Co2+ and Zn2+. EDTA did not affect the enzyme activity, whereas the presence of various oxidizing agents, reducing agents and some surfactants, reduced the enzyme activity. The enzyme was highly stable in the presence of some commercial detergent formulations. The values of Km and Vmax, as calculated from the Lineweaver–Burk plot, were 2.2 mg/ml and 1429 U/ml, respectively.

Item Type: Article
Additional Information: Copyright of this article belongs to Elsevier science.
Uncontrolled Keywords: Alkaline; Lipase; Thermophilic; Bacillus sp; Purification; Thermostable
Subjects: Q Science > QD Chemistry
Depositing User: Dr. K.P.S.Sengar
Date Deposited: 13 Feb 2012 15:17
Last Modified: 09 Jan 2015 10:51
URI: http://crdd.osdd.net/open/id/eprint/923

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