Part:BBa_K258006:Experience

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Applications of BBa_K258006

Lipase activity To identify a secretion phenotype on solid medium, E. coli was grown at 25°C for 48 h on LAT (LB medium, 1.5% Bacto Agar, 0.5% tributylin). The phenotype was evident by the development of a halo due to the secreted lipase [12]. In addition, lipase activity was measured spectrophotometrically using p-nitrophenyl palmitate (pNPP) as a substrate [12]. Ten millimolar pNPP dissolved in acetonitrile was mixed with ethanol and 50 mM Tris-HCl (pH 8.5) to a final ratio of acetonitrile:ethanol:Tris-HCl of 1:4:95 (v/v/v). The reaction was started by adding 50 μl of culture supernatant to 200 μl of reaction mixture at 42°C, and absorbance at 420 nm was monitored with Hitachi Spectrophotometry for 20 min. The activity was measured by the increase of optical density (OD).

procedure from:

Export of recombinant proteins in Escherichia coli using ABC transporter with an attached lipase ABC transporter recognition domain (LARD) Chan Woo Chung1, Jinsun You1, Kyeongyeon Kim1, Yuseok Moon2, Hoeon Kim3 and Jung Hoon Ahn* Published: 29 January 2009 Microbial Cell Factories 2009, 8:11 doi:10.1186/1475-2859-8-11 Received: 18 November 2008 Accepted: 29 January 2009 This article is available from: http://www.microbialcellfactories.com/content/8/1/11

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