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

DIR@IMTECH: CSIR-Institute of Microbial Technology

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Title Purification and characterisation of a thermostable alkaline lipase from a new thermophilic Bacillus sp. RSJ-1
 
Creator Sharma, R
Soni, S.K
Vohra, R M
Gupta, L.K.
Gupta, J.K.
 
Subject QD Chemistry
 
Description 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.
 
Publisher Elsevier Science
 
Date 2002
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://crdd.osdd.net/open/923/1/vohra2002.pdf
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
 
Relation http://dx.doi.org/10.1016/S0032-9592(01)00316-8
http://crdd.osdd.net/open/923/