Fig. 3

LipA variants showing increased extracellular lipase activity (a) or increased extracellular lipase amount (b). B. subtilis TEB1030 producing the different LipA variants were cultivated for 6 h in a 48 well Flowerplate^®, and the relative extracellular specific lipase activity (U/mg) in the culture supernatant was calculated by normalizing the volume activity (U/ml) to the determined protein amount (mg/ml). The extracellular lipase activity was measured using pNPP as a substrate, and the extracellular protein amount was determined by ELISA using a specific polyclonal LipA antibody. LipA variants with significant (p < 0.05) increase in extracellular specific lipase activity (n = 9) are shown relative to wtLipA. wtLipA values were set to 1 (thick black line)