Part:BBa_K3051001
Candida Antarctica Lipase A
Comparative Enzyme Activity Assay
Figure 1. Relative Comparaison Lipase Activity Assay. Candida Antarctica Lipase A, Bacillus Subtilis Lipase and Compost Lipase enzyme activity were examined using p-Nitrophenol octanoate as a substrate.
Figure 1 compares enzyme activity of Candida Antarctica Lipase A BBa_K3051001, Bacillus Subtilis Lipase BBa_K3051005 and Compost Lipase BBa_K3051421. The lipase parts were tested and characterized using a p nitrophenol octanoate assay. The enzyme activity was determined by measuring change in light absorbance at a wavelength of 400nm, which directly correlated to the concentration of p-Nitrophenol (pNP), caused by lipolysis of p-Nitrophenol octanoate (pNPO) substrate. The data from the pNPO lipolysis assay defined the enzyme activity.
Protein 3D Structure Prediction
Candida Antartica Lipase A (CALA) is a lipase extracted from the yeast Candida Antartica - the enzyme has expressed function at extremely low temperatures. CALA shows high affinity to C8 p-Nitrophenol esters, compared to C4 and >C12 esters (where C denotes the number of carbons in the fatty acid tail bound to the p-Nitrophenol group.)
The protein has shown sensitivity toward phenylmethylsulfonyl fluoride and high alcohol contents, causing a drop in activity to as low as 50% compared to the control.[1]
The enzyme has a kDa of 49.3 and has been shown to hydrolyse p-Nitrophenol octanoate into p-Nitrophenol. The molar extinction coefficient in water has been calculated to be 54570 M-1cm-1 (using ProtoPram ExPaSy tool). A 3D structure of the enzyme has been calculated.</div>
Figure 2. Simulated 3D structure.Structure of Candida Antartica lipase simulated using Phyre 2 modelling.
Sources 1. Lan, Dong-Ming et al. “A novel cold-active lipase from Candida albicans: cloning, expression and characterization of the recombinant enzyme.” International journal of molecular sciences vol. 12,6 (2011): 3950-65. doi:10.3390/ijms12063950
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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