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Fig. 4 | Microbial Cell Factories

Fig. 4

From: Contribution of single amino acid and codon substitutions to the production and secretion of a lipase by Bacillus subtilis

Fig. 4

Combination of beneficial single amino acid mutations in LipA and effects on (a) extracellular lipase activity and amount, and (b) additive effect of two amino acid substitutions on amount of extracellular lipase. B. subtilis TEB1030 harboring 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 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. The relative extracellular lipase amount (dark grey bars) and relative extracellular specific lipase activity (light grey bars) are given relative to wtLipA. Respective wtLipA values were set to 1 (thick black line). Significant changes (p < 0.05) compared to wtLipA are marked with a *, significant changes compared to the single variants (p < 0.05) are marked with **

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